US20160251659A1

US20160251659A1 - MicroRNA-Based Methods and Compositions for the Diagnosis, Prognosis and Treatment of Breast Cancer

Info

Publication number: US20160251659A1
Application number: US15/155,479
Authority: US
Inventors: Carlo M. Croce; George A. Calin
Original assignee: Ohio State University Research Foundation
Current assignee: Ohio State University Research Foundation
Priority date: 2005-08-01
Filing date: 2016-05-16
Publication date: 2016-09-01
Also published as: WO2007016548A2; US20140147495A1; CN103882124B; JP2009505639A; JP2013230161A; JP2013116114A; CN103866018A; ES2545383T3; CN103866017A; WO2007016548A3; JP5489459B2; EP1937845A2; CN103866018B; CN101341259A; CN103820562A; CN103820562B; CN101341259B; EP1937845B1; CN102533966B; CA2617581A1

Abstract

The present invention provides novel methods and compositions for the diagnosis, prognosis and treatment of breast cancer. The invention also provides methods of identifying anti-breast cancer agents.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of U.S. Ser. No. 14/145,364 filed Dec. 31, 2013, still pending, which is a divisional application of U.S. Ser. No. 12/012,235, filed Jan. 31, 2008, now U.S. Pat. No. 8,658,370 issued Feb. 25, 2014, which entered the National Phase on Jan. 31, 2008, from the International PCT Application No. U.S. Ser. No. 06/029,889, filed Jul. 31, 2006, which claims the benefit of U.S. Provisional Application No. 60/704,464, filed Aug. 1, 2005. The disclosures of each of the aforementioned applications are incorporated herein by reference for all purposes.

GOVERNMENT SUPPORT

This invention was supported by a grant under Program Project Grant P01CA76259, P01CA81534, and P30CA56036 from the National Cancer Institute. The Government has certain rights in this invention.

BACKGROUND OF THE INVENTION

Breast cancer is a significant health problem for women in the United States and throughout the world. Although advances have been made in the detection and treatment of the disease, breast cancer remains the second leading cause of cancer-related deaths in women, affecting more than 180,000 women in the United States each year. For women in North America, the life-time odds of getting breast cancer are now one in eight.
No universally successful method for the treatment or prevention of breast cancer is currently available. Management of breast cancer currently relies on a combination of early diagnosis (for example, through routine breast screening procedures) and aggressive treatment, which may include one or more of a variety of treatments, such as surgery, radiotherapy, chemotherapy and hormone therapy. The course of treatment for a particular breast cancer is often selected based on a variety of prognostic parameters including an analysis of specific tumor markers.
Although the discovery of BRCA1 and BRCA2 were important steps in identifying key genetic factors involved in breast cancer, it has become clear that mutations in BRCA1 and BRCA2 account for only a fraction of inherited susceptibility to breast cancer. In spite of considerable research into therapies for breast cancer, breast cancer remains difficult to diagnose and treat effectively, and the high mortality observed in breast cancer patients indicates that improvements are needed in the diagnosis, treatment and prevention of the disease.
MicroRNAs are a class of small, non-coding RNAs that control gene expression by hybridizing to and triggering either translational repression or, less frequently, degradation of a messenger RNA (mRNA) target. The discovery and study of miRNAs has revealed miRNA-mediated gene regulatory mechanisms that play important roles in organismal development and various cellular processes, such as cell differentiation, cell growth and cell death. Recent studies suggest that aberrant expression of particular miRNAs may be involved in human diseases, such as neurological disorders and cancer. In particular, misexpression of miR-16-1 and/or miR-15a has been found in human chronic lymphocytic leukemias.
The development and use of microarrays containing all known human microRNAs has permitted a simultaneous analysis of the expression of every miRNA in a sample. These microRNA microarrays have not only been used to confirm that miR-16-1 is deregulated in human CLL cells, but also to generate miRNA expression signatures that are associated with well-defined clinico-pathological features of human CLL.
The use of microRNA microarrays to identify a group of microRNAs, which are differentially-expressed between normal cells and breast cancer cells (for example, an expression signature or expression profile), may help pinpoint specific miRNAs that are involved in breast cancer. Furthermore, the identification of putative targets of these miRNAs may help to unravel their pathogenic role. The present invention provides novel methods and compositions for the diagnosis, prognosis and treatment of breast cancer.

SUMMARY OF THE INVENTION

The present invention is based, in part, on the identification of a breast cancer-specific signature of miRNAs that are differentially-expressed in breast cancer cells, relative to normal control cells.
Accordingly, embodiments of the invention encompass methods of diagnosing whether a subject has, or is at risk for developing, breast cancer, comprising measuring the level of at least one miR gene product in a test sample from the subject and comparing the level of the miR gene product in the test sample to the level of a corresponding miR gene product in a control sample. An alteration (for example, an increase, a decrease) in the level of the miR gene product in the test sample, relative to the level of a corresponding miR gene product in a control sample, is indicative of the subject either having, or being at risk for developing, breast cancer. In certain embodiments, the at least one miR gene product is selected from the group consisting of miR-125b-1, miR125b-2, miR-145, miR-21, miR-155, miR-10b and combinations thereof.
The level of the at least one miR gene product can be measured using a variety of techniques that are well known to those of skill in the art. In one embodiment, the level of the at least one miR gene product is measured using Northern blot analysis. In another embodiment, the level of the at least one miR gene product is measured by reverse transcribing RNA from a test sample obtained from the subject to provide a set of target oligodeoxynucleotides, hybridizing the target oligodeoxynucleotides to a microarray that comprises miRNA-specific probe oligonucleotides to provide a hybridization profile for the test sample, and comparing the test sample hybridization profile to a hybridization profile generated from a control sample. An alteration in the signal of at least one miRNA in the test sample relative to the control sample is indicative of the subject either having, or being at risk for developing, breast cancer. In a particular embodiment, the microarray comprises miRNA-specific probe oligonucleotides for a substantial portion of the human miRNome. In a further embodiment, the microarray comprises miRNA-specific probe oligonucleotides for one or more miRNAs selected from the group consisting of miR-145, miR-21, miR-155, miR-10b, miR-009-1 (miR131-1), miR-34 (miR-170), miR-102 (miR-29b), miR-123 (miR-126), miR-140-as, miR-125a, miR-125b-1, miR-125b-2, miR-194, miR-204, miR-213, let-7a-2, let-7a-3, let-7d (let-7d-v1), let-7f-2, let-7i (let-7d-v2), miR-101-1, miR-122a, miR-128b, miR-136, miR-143, miR-149, miR-191, miR-196-1, miR-196-2, miR-202, miR-203, miR-205, miR-206, miR-210 and combinations thereof.
Embodiments of the invention also provide methods of diagnosing a breast cancer associated with one or more prognostic markers, comprising measuring the level of at least one miR gene product in a breast cancer test sample from a subject and comparing the level of the at least one miR gene product in the breast cancer test sample to the level of a corresponding miR gene product in a control sample. The breast cancer can be associated with one or more adverse prognostic markers associated with breast cancer, such as, but not limited to, estrogen receptor expression, progesterone receptor expression, positive lymph node metastasis, high proliferative index, detectable p53 expression, advanced tumor stage, and high vascular invasion. In one embodiment, the level of the at least one miR gene product is measured by reverse transcribing RNA from a test sample obtained from the subject to provide a set of target oligodeoxynucleotides, hybridizing the target oligodeoxynucleotides to a microarray that comprises miRNA-specific probe oligonucleotides to provide a hybridization profile for the test sample, and comparing the test sample hybridization profile to a hybridization profile generated from a control sample. An alteration in the signal of at least one miRNA in the test sample relative to the control sample is indicative of the subject either having, or being at risk for developing, a breast cancer associated with the one or more prognostic markers. In a particular embodiment, the microarray comprises at least one miRNA-specific probe oligonucleotide for a miRNA selected from the group consisting of miR-26a, miR-26b, miR-102 (miR-29b), miR-30a-5p, miR-30b, miR-30c, miR-30d, miR-185, miR-191, miR-206, miR-212, let-7c, miR-9-2, miR-15-a, miR-21, miR-30a-s, miR-133a-1, miR-137, miR-153-2, miR-154, miR-181a, miR-203, miR-213, let-7f-1, let-7a-3, let-7a-2, miR-9-3, miR-10b, miR-27a, miR-29a, miR-123, miR-205, let-7d, miR-145, miR-16a, miR-128b and combinations thereof.
Embodiments of the invention also encompass methods of treating breast cancer in a subject, wherein at least one miR gene product is de-regulated (for example, down-regulated, up-regulated) in the cancer cells of the subject. When the at least one isolated miR gene product is down-regulated in the breast cancer cells, the method comprises administering an effective amount of the at least one isolated miR gene product, such that proliferation of cancer cells in the subject is inhibited. In one embodiment, the method comprises administering an effective amount of the at least one isolated miR gene product, provided that the miR gene is not miR-15a or miR-16-1, such that proliferation of cancer cells in the subject is inhibited. When the at least one isolated miR gene product is up-regulated in the cancer cells, the method comprises administering to the subject an effective amount of at least one compound for inhibiting expression of the at least one miR gene, such that proliferation of breast cancer cells is inhibited.
In related embodiments, the invention provides methods of treating breast cancer in a subject, comprising determining the amount of at least one miR gene product in breast cancer cells from the subject, relative to control cells. If expression of the miR gene product is deregulated in breast cancer cells, the methods further comprise altering the amount of the at least one miR gene product expressed in the breast cancer cells. If the amount of the miR gene product expressed in the cancer cells is less than the amount of the miR gene product expressed in control cells, the method comprises administering an effective amount of at least one isolated miR gene product. In one embodiment, the miR gene product is not miR-15a or miR-16-1. If the amount of the miR gene product expressed in the cancer cells is greater than the amount of the miR gene product expressed in control cells, the method comprises administering to the subject an effective amount of at least one compound for inhibiting expression of the at least one miR gene. In one embodiment, the miR gene product is not miR-15a or miR-16-1.
The invention further provides pharmaceutical compositions for treating breast cancer. In one embodiment, the pharmaceutical compositions comprise at least one isolated miR gene product and a pharmaceutically-acceptable carrier. In a particular embodiment, the at least one miR gene product corresponds to a miR gene product that has a decreased level of expression in breast cancer cells relative to suitable control cells. In certain embodiments the isolated miR gene product is selected from the group consisting of miR-145, miR-10b, miR-123 (miR-126), miR-140-as, miR-125a, miR-125b-1, miR-125b-2, miR-194, miR-204, let-7a-2, let-7a-3, let-7d (let-7d-v1), let-7f-2, miR-101-1, miR-143 and combinations thereof.
In another embodiment, the pharmaceutical compositions of the invention comprise at least one miR expression inhibition compound. In a particular embodiment, the at least one miR expression inhibition compound is specific for a miR gene whose expression is greater in breast cancer cells than control cells. In certain embodiments, the miR expression inhibition compound is specific for one or more miR gene products selected from the group consisting of miR-21, miR-155, miR-009-1 (miR131-1), miR-34 (miR-170), miR-102 (miR-29b), miR-213, let-7i (let-7d-v2), miR-122a, miR-128b, miR-136, miR-149, miR-191, miR-196-1, miR-196-2, miR-202, miR-203, miR-206, miR-210, miR-213 and combinations thereof.
Embodiments of the invention also encompass methods of identifying an anti-breast cancer agent, comprising providing a test agent to a cell and measuring the level of at least one miR gene product in the cell. In one embodiment, the method comprises providing a test agent to a cell and measuring the level of at least one miR gene product associated with decreased expression levels in breast cancer cells. An increase in the level of the miR gene product in the cell, relative to a suitable control cell, is indicative of the test agent being an anti-breast cancer agent. In a particular embodiment, the at least one miR gene product associated with decreased expression levels in breast cancer cells is selected from the group consisting of miR-145, miR-10b, miR-123 (miR-126), miR-140-as, miR-125a, miR-125b-1, miR-125b-2, miR-194, miR-204, let-7a-2, let-7a-3, let-7d (let-7d-v1), let-7f-2, miR-101-1, miR-143 and combinations thereof.
In other embodiments the method comprises providing a test agent to a cell and measuring the level of at least one miR gene product associated with increased expression levels in breast cancer cells. A decrease in the level of the miR gene product in the cell, relative to a suitable control cell, is indicative of the test agent being an anti-breast cancer agent. In a particular embodiment, at least one miR gene product associated with increased expression levels in breast cancer cells is selected from the group consisting of miR-21, miR-155, miR-009-1 (miR131-1), miR-34 (miR-170), miR-102 (miR-29b), miR-213, let-7i (let-7d-v2), miR-122a, miR-128b, miR-136, miR-149, miR-191, miR-196-1, miR-196-2, miR-202, miR-203, miR-206, miR-210, miR-213 and combinations thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

FIG. 1 depicts a tree generated by cluster analysis showing a separation of breast cancer from normal tissues on the basis of differential microRNA expression (P<0.05). The bar at the bottom of the figure indicates the group of cancer (red) or normal breast tissues (yellow).

FIG. 2 is a graph depicting the probability (0.0 to 1.0) of each sample being a cancerous or normal tissue based on PAM analysis. All breast cancer and normal tissues were correctly predicted by the miR signature shown in Table 2.

FIG. 3A is a Northern blot depicting the expression level of miR-125b, using a miR-125b complementary probe, in a normal sample, as well as several tumor samples from breast cancer patients (P). The U6 probe was used for normalization of expression levels for each sample.

FIG. 3B is a Northern blot depicting the expression level of miR-145, using a miR-145 complementary probe, in a normal sample, as well as several tumor samples from breast cancer patients (P). The U6 probe was used for normalization of expression levels for each sample.

FIG. 3C is a Northern blot depicting the expression level of miR-21, using a miR-21 complementary probe, in a normal sample, as well as several tumor samples from breast cancer patients (labeled as numbered patients). The U6 probe was used for normalization of expression levels for each sample.

FIG. 3D is a Northern blot depicting the expression levels of microRNAs miR-125b, miR-145 and miR-21 in various breast cancer cell lines. The expression level of each microRNA was also determined in a sample from normal tissues. The U6 probe was used for normalization of expression levels for each sample.

FIG. 4A is a table listing miRNAs that are differentially-expressed in breast cancer samples associated with the presence (ER+) or absence (ER−) of estrogen receptor.

FIG. 4B is a table listing miRNAs that are differentially-expressed in breast cancer samples associated with the presence (PR+) or absence (PR−) of progesterone receptor.

FIG. 4C is a table listing miRNAs that are differentially-expressed in breast cancer samples associated with stage 1 (pT1) or stage 2 or 3 (pT2-3) tumors.

FIG. 4D is a table listing miRNAs that are differentially-expressed in breast cancer samples associated with the presence (pN0) or absence (pN10+) of lymph node metastasis.

FIG. 4E is a table listing miRNAs that are differentially-expressed in breast cancer samples associated with the presence or absence of vascular invasion.

FIG. 4F is a table listing miRNAs that are differentially-expressed in breast cancer samples associated with a high (MIB-1>30) or low (MIB-1<20) proliferative index (PI).

FIG. 4G is a table listing miRNAs that are differentially-expressed in breast cancer samples associated with positive (p53+) or negative (p53−) immunostaining of p53.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based, in part, on the identification of particular miRNAs whose expression is altered in breast cancer cells relative to normal control cells, and microRNAs whose expression is altered in breast cancer cells associated with particular prognostic features, relative to breast cancer cells lacking such features.
As used herein interchangeably, a “miR gene product,” “microRNA,” “miR,” or “miRNA” refers to the unprocessed or processed RNA transcript from an miR gene. As the miR gene products are not translated into protein, the term “miR gene products” does not include proteins. The unprocessed miR gene transcript is also called an “miR precursor,” and typically comprises an RNA transcript of about 70-100 nucleotides in length. The miR precursor can be processed by digestion with an RNAse (for example, Dicer, Argonaut, or RNAse III, for example, E. coli RNAse III)) into an active 19-25 nucleotide RNA molecule. This active 19-25 nucleotide RNA molecule is also called the “processed” miR gene transcript or “mature” miRNA.
The active 19-25 nucleotide RNA molecule can be obtained from the miR precursor through natural processing routes (for example, using intact cells or cell lysates) or by synthetic processing routes (for example, using isolated processing enzymes, such as isolated Dicer, Argonaut, or RNAase III). It is understood that the active 19-25 nucleotide RNA molecule can also be produced directly by biological or chemical synthesis, without having been processed from the miR precursor.
The sequences of 187 miR gene products are provided in Table 1. All nucleic acid sequences herein are given in the 5′ to 3′ direction. In addition, genes are represented by italics, and gene products are represented by normal type; for example, mir-17 is the gene and miR-17 is the gene product.
The present invention encompasses methods of diagnosing whether a subject has, or is at risk for developing, breast cancer, comprising measuring the level of at least one miR gene product in a test sample from the subject and comparing the level of the miR gene product in the test sample to the level of a corresponding miR gene product in a control sample. As used herein, a “subject” can be any mammal that has, or is suspected of having, breast cancer. In a particular embodiment, the subject is a human who has, or is suspected of having, breast cancer.
The breast cancer can be any form of breast cancer and may be associated with one or more prognostic markers or features, including, but not limited to, estrogen receptor expression, progesterone receptor expression, lymph node metastasis, high proliferative index, detectable p53 expression, advanced tumor stage, and high vascular invasion. The prognostic marker can be associated with an adverse or negative prognosis, or it may be associated with a good or positive prognosis.

TABLE 1

Human miR Gene Product Sequences

	Precursor Sequence	SEQ ID
Name	(5′ to 3′)*	NO.

hsa-let-	CACTGTGGGATGAGGTAGTAGGTT	1
7a-1-prec	GTATAGTTTTAGGGTCACACCCAC
	CACTGGGAGATAACTATACAATCT
	ACTGTCTTTCCTAACGTG

hsa-let-	AGGTTGAGGTAGTAGGTTGTATAG	2
7a-2-prec	TTTAGAATTACATCAAGGGAGATA
	ACTGTACAGCCTCCTAGCTTTCCT

hsa-let-	GGGTGAGGTAGTAGGTTGTATAGT	3
7a-3-prec	TTGGGGCTCTGCCCTGCTATGGGA
	TAACTATACAATCTACTGTCTTTC
	CT

hsa-let-	GTGACTGCATGCTCCCAGGTTGAG	4
7a-4-prec	GTAGTAGGTTGTATAGTTTAGAAT
	TACACAAGGGAGATAACTGTACAG
	CCTCCTAGCTTTCCTTGGGTCTTG
	CACTAAACAAC

hsa-let-	GGCGGGGTGAGGTAGTAGGTTGTG	5
7b-prec	TGGTTTCAGGGCAGTGATGTTGCC
	CCTCGGAAGATAACTATACAACCT
	ACTGCCTTCCCTG

hsa-let-	GCATCCGGGTTGAGGTAGTAGGTT	6
7c-prec	GTATGGTTTAGAGTTACACCCTGG
	GAGTTAACTGTACAACCTTCTAGC
	TTTCCTTGGAGC

hsa-let-	CCTAGGAAGAGGTAGTAGGTTGCA	7
7d-prec	TAGTTTTAGGGCAGGGATTTTGCC
	CACAAGGAGGTAACTATACGACCT
	GCTGCCTTTCTTAGG

hsa-let-	CTAGGAAGAGGTAGTAGTTTGCAT	8
7d-v1-prec	AGTTTTAGGGCAAAGATTTTGCCC
	ACAAGTAGTTAGCTATACGACCTG
	CAGCCTTTTGTAG

hsa-let-	CTGGCTGAGGTAGTAGTTTGTGCT	9
7d-v2-prec	GTTGGTCGGGTTGTGACATTGCCC
	GCTGTGGAGATAACTGCGCAAGCT
	ACTGCCTTGCTAG

hsa-let-	CCCGGGCTGAGGTAGGAGGTTGTA	10
7e-prec	TAGTTGAGGAGGACACCCAAGGAG
	ATCACTATACGGCCTCCTAGCTTT
	CCCCAGG

hsa-let-	TCAGAGTGAGGTAGTAGATTGTAT	11
7f-1-prec	AGTTGTGGGGTAGTGATTTTACCC
	TGTTCAGGAGATAACTATACAATC
	TATTGCCTTCCCTGA

hsa-let-	CTGTGGGATGAGGTAGTAGATTGT	12
7f-2-prec	ATAGTTGTGGGGTAGTGATTTTAC
	CCTGTTCAGGAGATAACTATACAA
	TCTATTGCCTTCCCTGA

hsa-let-	CTGTGGGATGAGGTAGTAGATTGT	13
7f-2-prec	ATAGTTTTAGGGTCATACCCCATC
	TTGGAGATAACTATACAGTCTACT
	GTCTTTCCCACGG

hsa-let-	TTGCCTGATTCCAGGCTGAGGTAG	14
7g-prec	TAGTTTGTACAGTTTGAGGGTCTA
	TGATACCACCCGGTACAGGAGATA
	ACTGTACAGGCCACTGCCTTGCCA
	GGAACAGCGCGC

hsa-let-	CTGGCTGAGGTAGTAGTTTGTGCT	15
7i-prec	GTTGGTCGGGTTGTGACATTGCCC
	GCTGTGGAGATAACTGCGCAAGCT
	ACTGCCTTGCTAG

hsa-mir-	ACCTACTCAGAGTACATACTTCTT	16
001b-1-prec	TATGTACCCATATGAACATACAAT
	GCTATGGAATGTAAAGAAGTATGT
	ATTTTTGGTAGGC

hsa-mir-	CAGCTAACAACTTAGTAATACCTA	17
001b-1-prec	CTCAGAGTACATACTTCTTTATGT
	ACCCATATGAACATACAATGCTAT
	GGAATGTAAAGAAGTATGTATTTT
	TGGTAGGCAATA

hsa-mir-	GCCTGCTTGGGAAACATACTTCTT	18
001b-2-prec	TATATGCCCATATGGACCTGCTAA
	GCTATGGAATGTAAAGAAGTATGT
	ATCTCAGGCCGGG

hsa-mir-	TGGGAAACATACTTCTTTATATGC	19
001b-prec	CCATATGGACCTGCTAAGCTATGG
	AATGTAAAGAAGTATGTATCTCA

hsa-mir-	ACCTACTCAGAGTACATACTTCTT	20
001d-prec	TATGTACCCATATGAACATACAAT
	GCTATGGAATGTAAAGAAGTATGT
	ATTTTTGGTAGGC

hsa-mir-	TGGATGTTGGCCTAGTTCTGTGTG	21
007-1	GAAGACTAGTGATTTTGTTGTTTT
	TAGATAACTAAATCGACAACAAAT
	CACAGTCTGCCATATGGCACAGGC
	CATGCCTCTACA

hsa-mir-	TTGGATGTTGGCCTAGTTCTGTGT	22
007-1-prec	GGAAGACTAGTGATTTTGTTGTTT
	TTAGATAACTAAATCGACAACAAA
	TCACAGTCTGCCATATGGCACAGG
	CCATGCCTCTACAG

hsa-mir-	CTGGATACAGAGTGGACCGGCTGG	23
007-2	CCCCATCTGGAAGACTAGTGATTT
	TGTTGTTGTCTTACTGCGCTCAAC
	AACAAATCCCAGTCTACCTAATGG
	TGCCAGCCATCGCA

hsa-mir-	CTGGATACAGAGTGGACCGGCTGG	24
007-2-prec	CCCCATCTGGAAGACTAGTGATTT
	TGTTGTTGTCTTACTGCGCTCAAC
	AACAAATCCCAGTCTACCTAATGG
	TGCCAGCCATCGCA

hsa-mir-	AGATTAGAGTGGCTGTGGTCTAGT	25
007-3	GCTGTGTGGAAGACTAGTGATTTT
	GTTGTTCTGATGTACTACGACAAC
	AAGTCACAGCCGGCCTCATAGCGC
	AGACTCCCTTCGAC

hsa-mir-	AGATTAGAGTGGCTGTGGTCTAGT	26
007-3-prec	GCTGTGTGGAAGACTAGTGATTTT
	GTTGTTCTGATGTACTACGACAAC
	AAGTCACAGCCGGCCTCATAGCGC
	AGACTCCCTTCGAC

hsa-mir-	CGGGGTTGGTTGTTATCTTTGGTT	27
009-1	ATCTAGCTGTATGAGTGGTGTGGA
	GTCTTCATAAAGCTAGATAACCGA
	AAGTAAAAATAACCCCA

hsa-mir-	GGAAGCGAGTTGTTATCTTTGGTT	28
009-2	ATCTAGCTGTATGAGTGTATTGGT
	CTTCATAAAGCTAGATAACCGAAA
	GTAAAAACTCCTTCA

hsa-mir-	GGAGGCCCGTTTCTCTCTTTGGTT	29
009-3	ATCTAGCTGTATGAGTGCCACAGA
	GCCGTCATAAAGCTAGATAACCGA
	AAGTAGAAATGATTCTCA

hsa-mir-	GATCTGTCTGTCTTCTGTATATAC	30
010a-prec	CCTGTAGATCCGAATTTGTGTAAG
	GAATTTTGTGGTCACAAATTCGTA
	TCTAGGGGAATATGTAGTTGACAT
	AAACACTCCGCTCT

hsa-mir-	CCAGAGGTTGTAACGTTGTCTATA	31
010b-prec	TATACCCTGTAGAACCGAATTTGT
	GTGGTATCCGTATAGTCACAGATT
	CGATTCTAGGGGAATATATGGTCG
	ATGCAAAAACTTCA

hsa-mir-	GCGCGAATGTGTGTTTAAAAAAAA	32
015a-2-prec	TAAAACCTTGGAGTAAAGTAGCAG
	CACATAATGGTTTGTGGATTTTGA
	AAAGGTGCAGGCCATATTGTGCTG
	CCTCAAAAATAC

hsa-mir-	CCTTGGAGTAAAGTAGCAGCACAT	33
015a-prec	AATGGTTTGTGGATTTTGAAAAGG
	TGCAGGCCATATTGTGCTGCCTCA
	AAAATACAAGG

hsa-mir-	CTGTAGCAGCACATCATGGTTTAC	34
015b-prec	ATGCTACAGTCAAGATGCGAATCA
	TTATTTGCTGCTCTAG

hsa-mir-	TTGAGGCCTTAAAGTACTGTAGCA	35
015b-prec	GCACATCATGGTTTACATGCTACA
	GTCAAGATGCGAATCATTATTTGC
	TGCTCTAGAAATTTAAGGAAATTC
	AT

hsa-mir-	GTCAGCAGTGCCTTAGCAGCACGT	36
016a-chr13	AAATATTGGCGTTAAGATTCTAAA
	ATTATCTCCAGTATTAACTGTGCT
	GCTGAAGTAAGGTTGAC

hsa-mir-	GTTCCACTCTAGCAGCACGTAAAT	37
016b-chr3	ATTGGCGTAGTGAAATATATATTA
	AACACCAATATTACTGTGCTGCTT
	TAGTGTGAC

hsa-mir-	GCAGTGCCTTAGCAGCACGTAAAT	38
016-prec-13	ATTGGCGTTAAGATTCTAAAATTA
	TCTCCAGTATTAACTGTGCTGCTG
	AAGTAAGGT

hsa-mir-	GTCAGAATAATGTCAAAGTGCTTA	39
017-prec	CAGTGCAGGTAGTGATATGTGCAT
	CTACTGCAGTGAAGGCACTTGTAG
	CATTATGGTGAC

hsa-mir-	TGTTCTAAGGTGCATCTAGTGCAG	40
018-prec	ATAGTGAAGTAGATTAGCATCTAC
	TGCCCTAAGTGCTCCTTCTGGCA

hsa-mir-	TTTTTGTTCTAAGGTGCATCTAGT	41
018-prec-13	GCAGATAGTGAAGTAGATTAGCAT
	CTACTGCCCTAAGTGCTCCTTCTG
	GCATAAGAA

hsa-mir-	GCAGTCCTCTGTTAGTTTTGCATA	42
019a-prec	GTTGCACTACAAGAAGAATGTAGT
	TGTGCAAATCTATGCAAAACTGAT
	GGTGGCCTGC

hsa-mir-	CAGTCCTCTGTTAGTTTTGCATAG	43
019a-prec-13	TTGCACTACAAGAAGAATGTAGTT
	GTGCAAATCTATGCAAAACTGATG
	GTGGCCTG

hsa-mir-	CACTGTTCTATGGTTAGTTTTGCA	44
019b-1-prec	GGTTTGCATCCAGCTGTGTGATAT
	TCTGCTGTGCAAATCCATGCAAAA
	CTGACTGTGGTAGTG

hsa-mir-	ACATTGCTACTTACAATTAGTTTT	45
019b-2-prec	GCAGGTTTGCATTTCAGCGTATAT
	ATGTATATGTGGCTGTGCAAATCC
	ATGCAAAACTGATTGTGATAATGT

hsa-mir-	TTCTATGGTTAGTTTTGCAGGTTT	46
019b-prec-13	GCATCCAGCTGTGTGATATTCTGC
	TGTGCAAATCCATGCAAAACTGAC
	TGTGGTAG

hsa-mir-	TTACAATTAGTTTTGCAGGTTTGC	47
019b-prec-X	ATTTCAGCGTATATATGTATATGT
	GGCTGTGCAAATCCATGCAAAACT
	GATTGTGAT

hsa-mir-	GTAGCACTAAAGTGCTTATAGTGC	48
020-prec	AGGTAGTGTTTAGTTATCTACTGC
	ATTATGAGCACTTAAAGTACTGC

hsa-mir-	TGTCGGGTAGCTTATCAGACTGAT	49
021-prec	GTTGACTGTTGAATCTCATGGCAA
	CACCAGTCGATGGGCTGTCTGACA

hsa-mir-	ACCTTGTCGGGTAGCTTATCAGAC	50
021-prec-17	TGATGTTGACTGTTGAATCTCATG
	GCAACACCAGTCGATGGGCTGTCT
	GACATTTTG

hsa-mir-	GGCTGAGCCGCAGTAGTTCTTCAG	51
022-prec	TGGCAAGCTTTATGTCCTGACCCA
	GCTAAAGCTGCCAGTTGAAGAACT
	GTTGCCCTCTGCC

hsa-mir-	GGCCGGCTGGGGTTCCTGGGGATG	52
023a-prec	GGATTTGCTTCCTGTCACAAATCA
	CATTGCCAGGGATTTCCAACCGAC
	C

hsa-mir-	CTCAGGTGCTCTGGCTGCTTGGGT	53
023b-prec	TCCTGGCATGCTGATTTGTGACTT
	AAGATTAAAATCACATTGCCAGGG
	ATTACCACGCAACCACGACCTTGG
	C

hsa-mir-	CCACGGCCGGCTGGGGTTCCTGGG	54
023-prec-19	GATGGGATTTGCTTCCTGTCACAA
	ATCACATTGCCAGGGATTTCCAAC
	CGACCCTGA

hsa-mir-	CTCCGGTGCCTACTGAGCTGATAT	55
024-1-prec	CAGTTCTCATTTTACACACTGGCT
	CAGTTCAGCAGGAACAGGAG

hsa-mir-	CTCTGCCTCCCGTGCCTACTGAGC	56
024-2-prec	TGAAACACAGTTGGTTTGTGTACA
	CTGGCTCAGTTCAGCAGGAACAGG
	G

hsa-mir-	CCCTGGGCTCTGCCTCCCGTGCCT	57
024-prec-19	ACTGAGCTGAAACACAGTTGGTTT
	GTGTACACTGGCTCAGTTCAGCAG
	GAACAGGGG

hsa-mir-	CCCTCCGGTGCCTACTGAGCTGAT	58
024-prec-9	ATCAGTTCTCATTTTACACACTGG
	CTCAGTTCAGCAGGAACAGCATC

hsa-mir-	GGCCAGTGTTGAGAGGCGGAGACT	59
025-prec	TGGGCAATTGCTGGACGCTGCCCT
	GGGCATTGCACTTGTCTCGGTCTG
	ACAGTGCCGGCC

hsa-mir-	AGGCCGTGGCCTCGTTCAAGTAAT	60
026a-prec	CCAGGATAGGCTGTGCAGGTCCCA
	ATGGCCTATCTTGGTTACTTGCAC
	GGGGACGCGGGCCT

hsa-mir-	CCGGGACCCAGTTCAAGTAATTCA	61
026b-prec	GGATAGGTTGTGTGCTGTCCAGCC
	TGTTCTCCATTACTTGGCTCGGGG
	ACCGG

hsa-mir-	CTGAGGAGCAGGGCTTAGCTGCTT	62
027a-prec	GTGAGCAGGGTCCACACCAAGTCG
	TGTTCACAGTGGCTAAGTTCCGCC
	CCCCAG

hsa-mir-	AGGTGCAGAGCTTAGCTGATTGGT	63
027b-prec	GAACAGTGATTGGTTTCCGCTTTG
	TTCACAGTGGCTAAGTTCTGCACC
	T

hsa-mir-	ACCTCTCTAACAAGGTGCAGAGCT	64
027b-prec	TAGCTGATTGGTGAACAGTGATTG
	GTTTCCGCTTTGTTCACAGTGGCT
	AAGTTCTGCACCTGAAGAGAAGGT
	G

hsa-mir-	CCTGAGGAGCAGGGCTTAGCTGCT	65
027-prec-19	TGTGAGCAGGGTCCACACCAAGTC
	GTGTTCACAGTGGCTAAGTTCCGC
	CCCCCAGG

hsa-mir-	GGTCCTTGCCCTCAAGGAGCTCAC	66
028-prec	AGTCTATTGAGTTACCTTTCTGAC
	TTTCCCACTAGATTGTGAGCTCCT
	GGAGGGCAGGCACT

hsa-mir-	CCTTCTGTGACCCCTTAGAGGATG	67
029a-2	ACTGATTTCTTTTGGTGTTCAGAG
	TCAATATAATTTTCTAGCACCATC
	TGAAATCGGTTATAATGATTGGGG
	AAGAGCACCATG

hsa-mir-	ATGACTGATTTCTTTTGGTGTTCA	68
029a-prec	GAGTCAATATAATTTTCTAGCACC
	ATCTGAAATCGGTTAT

hsa-mir-	ACCACTGGCCCATCTCTTACACAG	69
029c-prec	GCTGACCGATTTCTCCTGGTGTTC
	AGAGTCTGTTTTTGTCTAGCACCA
	TTTGAAATCGGTTATGATGTAGGG
	GGAAAAGCAGCAGC

hsa-mir-	GCGACTGTAAACATCCTCGACTGG	70
030a-prec	AAGCTGTGAAGCCACAGATGGGCT
	TTCAGTCGGATGTTTGCAGCTGC

hsa-mir-	ATGTAAACATCCTACACTCAGCTG	71
030b-prec	TAATACATGGATTGGCTGGGAGGT
	GGATGTTTACGT

hsa-mir-	ACCAAGTTTCAGTTCATGTAAACA	72
030b-prec	TCCTACACTCAGCTGTAATACATG
	GATTGGCTGGGAGGTGGATGTTTA
	CTTCAGCTGACTTGGA

hsa-mir-	AGATACTGTAAACATCCTACACTC	73
030c-prec	TCAGCTGTGGAAAGTAAGAAAGCT
	GGGAGAAGGCTGTTTACTCTTTCT

hsa-mir-	GTTGTTGTAAACATCCCCGACTGG	74
030d-prec	AAGCTGTAAGACACAGCTAAGCTT
	TCAGTCAGATGTTTGCTGCTAC

hsa-mir-	GGAGAGGAGGCAAGATGCTGGCAT	75
031-prec	AGCTGTTGAACTGGGAACCTGCTA
	TGCCAACATATTGCCATCTTTCC

hsa-mir-	GGAGATATTGCACATTACTAAGTT	76
032-prec	GCATGTTGTCACGGCCTCAATGCA
	ATTTAGTGTGTGTGATATTTTC

hsa-mir-	GGGGGCCGAGAGAGGCGGGCGGCC	77
033b-prec	CCGCGGTGCATTGCTGTTGCATTG
	CACGTGTGTGAGGCGGGTGCAGTG
	CCTCGGCAGTGCAGCCCGGAGCCG
	GCCCCTGGCACCAC

hsa-mir-	CTGTGGTGCATTGTAGTTGCATTG	78
033-prec	CATGTTCTGGTGGTACCCATGCAA
	TGTTTCCACAGTGCATCACAG

hsa-mir-	GGCCAGCTGTGAGTGTTTCTTTGG	79
034-prec	CAGTGTCTTAGCTGGTTGTTGTGA
	GCAATAGTAAGGAAGCAATCAGCA
	AGTATACTGCCCTAGAAGTGCTGC
	ACGTTGTGGGGCCC

hsa-mir-	TCAGAATAATGTCAAAGTGCTTAC	80
091-prec-13	AGTGCAGGTAGTGATATGTGCATC
	TACTGCAGTGAAGGCACTTGTAGC
	ATTATGGTGA

hsa-mir-	CTTTCTACACAGGTTGGGATCGGT	81
092-prec-13 =	TGCAATGCTGTGTTTCTGTATGGT
092-1	ATTGCACTTGTCCCGGCCTGTTGA
	GTTTGG

hsa-mir-	TCATCCCTGGGTGGGGATTTGTTG	82
092-prec-X =	CATTACTTGTGTTCTATATAAAGT
092-2	ATTGCACTTGTCCCGGCCTGTGGA
	AGA

hsa-mir-	CTGGGGGCTCCAAAGTGCTGTTCG	83
093-prec-7.1 =	TGCAGGTAGTGTGATTACCCAACC
093-1	TACTGCTGAGCTAGCACTTCCCGA
	GCCCCCGG

hsa-mir-	CTGGGGGCTCCAAAGTGCTGTTCG	84
093-prec-7.2 =	TGCAGGTAGTGTGATTACCCAACC
093-2	TACTGCTGAGCTAGCACTTCCCGA
	GCCCCCGG

hsa-mir-	AACACAGTGGGCACTCAATAAATG	85
095-prec-4	TCTGTTGAATTGAAATGCGTTACA
	TTCAACGGGTATTTATTGAGCACC
	CACTCTGTG

hsa-mir-	TGGCCGATTTTGGCACTAGCACAT	86
096-prec-7	TTTTGCTTGTGTCTCTCCGCTCTG
	AGCAATCATGTGCAGTGCCAATAT
	GGGAAA

hsa-mir-	GTGAGGTAGTAAGTTGTATTGTTG	87
098-prec-X	TGGGGTAGGGATATTAGGCCCCAA
	TTAGAAGATAACTATACAACTTAC
	TACTTTCC

hsa-mir-	GGCACCCACCCGTAGAACCGACCT	88
099b-prec-19	TGCGGGGCCTTCGCCGCACACAAG
	CTCGTGTCTGTGGGTCCGTGTC

hsa-mir-	CCCATTGGCATAAACCCGTAGATC	89
099-prec-21	CGATCTTGTGGTGAAGTGGACCGC
	ACAAGCTCGCTTCTATGGGTCTGT
	GTCAGTGTG

hsa-mir-	AAGAGAGAAGATATTGAGGCCTGT	90
100-1/2-prec	TGCCACAAACCCGTAGATCCGAAC
	TTGTGGTATTAGTCCGCACAAGCT
	TGTATCTATAGGTATGTGTCTGTT
	AGGCAATCTCAC

hsa-mir-	CCTGTTGCCACAAACCCGTAGATC	91
100-prec-11	CGAACTTGTGGTATTAGTCCGCAC
	AAGCTTGTATCTATAGGTATGTGT
	CTGTTAGG

hsa-mir-	AGGCTGCCCTGGCTCAGTTATCAC	92
101-1/2-prec	AGTGCTGATGCTGTCTATTCTAAA
	GGTACAGTACTGTGATAACTGAAG
	GATGGCAGCCATCTTACCTTCCAT
	CAGAGGAGCCTCAC

hsa-mir-	TCAGTTATCACAGTGCTGATGCTG	93
101-prec	TCCATTCTAAAGGTACAGTACTGT
	GATAACTGA

hsa-mir-	TGCCCTGGCTCAGTTATCACAGTG	94
101-prec-1	CTGATGCTGTCTATTCTAAAGGTA
	CAGTACTGTGATAACTGAAGGATG
	GCA

hsa-mir-	TGTCCTTTTTCGGTTATCATGGTA	95
101-prec-9	CCGATGCTGTATATCTGAAAGGTA
	CAGTACTGTGATAACTGAAGAATG
	GTG

hsa-mir-	CTTCTGGAAGCTGGTTTCACATGG	96
102-prec-1	TGGCTTAGATTTTTCCATCTTTGT
	ATCTAGCACCATTTGAAATCAGTG
	TTTTAGGAG

hsa-mir-	CTTCAGGAAGCTGGTTTCATATGG	97
102-prec-7.1	TGGTTTAGATTTAAATAGTGATTG
	TCTAGCACCATTTGAAATCAGTGT
	TCTTGGGGG

hsa-mir-	CTTCAGGAAGCTGGTTTCATATGG	98
102-prec-7.2	TGGTTTAGATTTAAATAGTGATTG
	TCTAGCACCATTTGAAATCAGTGT
	TCTTGGGGG

hsa-mir-	TTGTGCTTTCAGCTTCTTTACAGT	99
103-2-prec	GCTGCCTTGTAGCATTCAGGTCAA
	GCAACATTGTACAGGGCTATGAAA
	GAACCA

hsa-mir-	TTGTGCTTTCAGCTTCTTTACAGT	100
103-prec-20	GCTGCCTTGTAGCATTCAGGTCAA
	GCAACATTGTACAGGGCTATGAAA
	GAACCA

hsa-mir-	TACTGCCCTCGGCTTCTTTACAGT	101
103-prec-5 =	GCTGCCTTGTTGCATATGGATCAA
103-1	GCAGCATTGTACAGGGCTATGAAG
	GCATTG

hsa-mir-	AAATGTCAGACAGCCCATCGACTG	102
104-prec-17	GTGTTGCCATGAGATTCAACAGTC
	AACATCAGTCTGATAAGCTACCCG
	ACAAGG

hsa-mir-	TGTGCATCGTGGTCAAATGCTCAG	103
105-prec-X.1 =	ACTCCTGTGGTGGCTGCTCATGCA
105-1	CCACGGATGTTTGAGCATGTGCTA
	CGGTGTCTA

hsa-mir-	TGTGCATCGTGGTCAAATGCTCAG	104
105-prec-X.2 =	ACTCCTGTGGTGGCTGCTCATGCA
105-2	CCACGGATGTTTGAGCATGTGCTA
	CGGTGTCTA

hsa-mir-	CCTTGGCCATGTAAAAGTGCTTAC	105
106-prec-X	AGTGCAGGTAGCTTTTTGAGATCT
	ACTGCAATGTAAGCACTTCTTACA
	TTACCATGG

hsa-mir-	CTCTCTGCTTTCAGCTTCTTTACA	106
107-prec-10	GTGTTGCCTTGTGGCATGGAGTTC
	AAGCAGCATTGTACAGGGCTATCA
	AAGCACAGA

hsa-mir-	CCTTAGCAGAGCTGTGGAGTGTGA	107
122a-prec	CAATGGTGTTTGTGTCTAAACTAT
	CAAACGCCATTATCACACTAAATA
	GCTACTGCTAGGC

hsa-mir-	AGCTGTGGAGTGTGACAATGGTGT	108
122a-prec	TTGTGTCCAAACTATCAAACGCCA
	TTATCACACTAAATAGCT

hsa-mir-	ACATTATTACTTTTGGTACGCGCT	109
123-prec	GTGACACTTCAAACTCGTACCGTG
	AGTAATAATGCGC

hsa-mir-	tccttcctCAGGAGAAAGGCCTCT	110
124a-1-prec	CTCTCCGTGTTCACAGCGGACCTT
	GATTTAAATGTCCATACAATTAAG
	GCACGCGGTGAATGCCAAGAATGG
	GGCT

hsa-mir-	AGGCCTCTCTCTCCGTGTTCACAG	111
124a-1-prec	CGGACCTTGATTTAAATGTCCATA
	CAATTAAGGCACGCGGTGAATGCC
	AAGAATGGGGCTG

hsa-mir-	ATCAAGATTAGAGGCTCTGCTCTC	112
124a-2-prec	CGTGTTCACAGCGGACCTTGATTT
	AATGTCATACAATTAAGGCACGCG
	GTGAATGCCAAGAGCGGAGCCTAC
	GGCTGCACTTGAAG

hsa-mir-	CCCGCCCCAGCCCTGAGGGCCCCT	113
124a-3-prec	CTGCGTGTTCACAGCGGACCTTGA
	TTTAATGTCTATACAATTAAGGCA
	CGCGGTGAATGCCAAGAGAGGCGC
	CTCCGCCGCTCCTT

hsa-mir-	TGAGGGCCCCTCTGCGTGTTCACA	114
124a-3-prec	GCGGACCTTGATTTAATGTCTATA
	CAATTAAGGCACGCGGTGAATGCC
	AAGAGAGGCGCCTCC

hsa-mir-	CTCTGCGTGTTCACAGCGGACCTT	115
124a-prec	GATTTAATGTCTATACAATTAAGG
	CACGCGGTGAATGCCAAGAG

hsa-mir-	CTCTCCGTGTTCACAGCGGACCTT	116
124b-prec	GATTTAATGTCATACAATTAAGGC
	ACGCGGTGAATGCCAAGAG

hsa-mir-	TGCCAGTCTCTAGGTCCCTGAGAC	117
125a-prec	CCTTTAACCTGTGAGGACATCCAG
	GGTCACAGGTGAGGTTCTTGGGAG
	CCTGGCGTCTGGCC

hsa-mir-	GGTCCCTGAGACCCTTTAACCTGT	118
125a-prec	GAGGACATCCAGGGTCACAGGTGA
	GGTTCTTGGGAGCCTGG

hsa-mir-	ACATTGTTGCGCTCCTCTCAGTCC	119
125b-1	CTGAGACCCTAACTTGTGATGTTT
	ACCGTTTAAATCCACGGGTTAGGC
	TCTTGGGAGCTGCGAGTCGTGCTT
	TTGCATCCTGGA

hsa-mir-	TGCGCTCCTCTCAGTCCCTGAGAC	120
125b-1	CCTAACTTGTGATGTTTACCGTTT
	AAATCCACGGGTTAGGCTCTTGGG
	AGCTGCGAGTCGTGCT

hsa-mir-	ACCAGACTTTTCCTAGTCCCTGAG	121
125b-2-prec	ACCCTAACTTGTGAGGTATTTTAG
	TAACATCACAAGTCAGGCTCTTGG
	GACCTAGGCGGAGGGGA

hsa-mir-	CCTAGTCCCTGAGACCCTAACTTG	122
125b-2-prec	TGAGGTATTTTAGTAACATCACAA
	GTCAGGCTCTTGGGACCTAGGC

hsa-mir-	CGCTGGCGACGGGACATTATTACT	123
126-prec	TTTGGTACGCGCTGTGACACTTCA
	AACTCGTACCGTGAGTAATAATGC
	GCCGTCCACGGCA

hsa-mir-	ACATTATTACTTTTGGTACGCGCT	124
126-prec	GTGACACTTCAAACTCGTACCGTG
	AGTAATAATGCGC

hsa-mir-	TGTGATCACTGTCTCCAGCCTGCT	125
127-prec	GAAGCTCAGAGGGCTCTGATTCAG
	AAAGATCATCGGATCCGTCTGAGC
	TTGGCTGGTCGGAAGTCTCATCAT
	C

hsa-mir-	CCAGCCTGCTGAAGCTCAGAGGGC	126
127-prec	TCTGATTCAGAAAGATCATCGGAT
	CCGTCTGAGCTTGGCTGGTCGG

hsa-mir-	TGAGCTGTTGGATTCGGGGCCGTA	127
128a-prec	GCACTGTCTGAGAGGTTTACATTT
	CTCACAGTGAACCGGTCTCTTTTT
	CAGCTGCTTC

hsa-mir-	GCCCGGCAGCCACTGTGCAGTGGG	128
128b-prec	AAGGGGGGCCGATACACTGTACGA
	GAGTGAGTAGCAGGTCTCACAGTG
	AACCGGTCTCTTTCCCTACTGTGT
	CACACTCCTAATGG

hsa-mir-	GTTGGATTCGGGGCCGTAGCACTG	129
128-prec	TCTGAGAGGTTTACATTTCTCACA
	GTGAACCGGTCTCTTTTTCAGC

hsa-mir-	TGGATCTTTTTGCGGTCTGGGCTT	130
129-prec	GCTGTTCCTCTCAACAGTAGTCAG
	GAAGCCCTTACCCCAAAAAGTATC
	TA

hsa-mir-	TGCTGCTGGCCAGAGCTCTTTTCA	131
130a-prec	CATTGTGCTACTGTCTGCACCTGT
	CACTAGCAGTGCAATGTTAAAAGG
	GCATTGGCCGTGTAGTG

hsa-mir-	gccaggaggcggGGTTGGTTGTTA	132
131-1-prec	TCTTTGGTTATCTAGCTGTATGAG
	TGGTGTGGAGTCTTCATAAAGCTA
	GATAACCGAAAGTAAAAATAACCC
	CATACACTGCGCAG

hsa-mir-	CACGGCGCGGCAGCGGCACTGGCT	133
131-3-prec	AAGGGAGGCCCGTTTCTCTCTTTG
	GTTATCTAGCTGTATGAGTGCCAC
	AGAGCCGTCATAAAGCTAGATAAC
	CGAAAGTAGAAATG

hsa-mir-	GTTGTTATCTTTGGTTATCTAGCT	134
131-prec	GTATGAGTGTATTGGTCTTCATAA
	AGCTAGATAACCGAAAGTAAAAAC

hsa-mir-	CCGCCCCCGCGTCTCCAGGGCAAC	135
132-prec	CGTGGCTTTCGATTGTTACTGTGG
	GAACTGGAGGTAACAGTCTACAGC
	CATGGTCGCCCCGCAGCACGCCCA
	CGCGC

hsa-mir-	GGGCAACCGTGGCTTTCGATTGTT	136
132-prec	ACTGTGGGAACTGGAGGTAACAGT
	CTACAGCCATGGTCGCCC

hsa-mir-	ACAATGCTTTGCTAGAGCTGGTAA	137
133a-1	AATGGAACCAAATCGCCTCTTCAA
	TGGATTTGGTCCCCTTCAACCAGC
	TGTAGCTATGCATTGA

hsa-mir-	GGGAGCCAAATGCTTTGCTAGAGC	138
133a-2	TGGTAAAATGGAACCAAATCGACT
	GTCCAATGGATTTGGTCCCCTTCA
	ACCAGCTGTAGCTGTGCATTGATG
	GCGCCG

hsa-mir-	GCTAGAGCTGGTAAAATGGAACCA	139
133-prec	AATCGCCTCTTCAATGGATTTGGT
	CCCCTTCAACCAGCTGTAGC

hsa-mir-	CAGGGTGTGTGACTGGTTGACCAG	140
134-prec	AGGGGCATGCACTGTGTTCACCCT
	GTGGGCCACCTAGTCACCAACCCT
	C

hsa-mir-	AGGGTGTGTGACTGGTTGACCAGA	141
134-prec	GGGGCATGCACTGTGTTCACCCTG
	TGGGCCACCTAGTCACCAACCCT

hsa-mir-	AGGCCTCGCTGTTCTCTATGGCTT	142
135-1-prec	TTTATTCCTATGTGATTCTACTGC
	TCACTCATATAGGGATTGGAGCCG
	TGGCGCACGGCGGGGACA

hsa-mir-	AGATAAATTCACTCTAGTGCTTTA	143
135-2-prec	TGGCTTTTTATTCCTATGTGATAG
	TAATAAAGTCTCATGTAGGGATGG
	AAGCCATGAAATACATTGTGAAAA
	ATCA

hsa-mir-	CTATGGCTTTTTATTCCTATGTGA	144
135-prec	TTCTACTGCTCACTCATATAGGGA
	TTGGAGCCGTGG

hsa-mir-	TGAGCCCTCGGAGGACTCCATTTG	145
136-prec	TTTTGATGATGGATTCTTATGCTC
	CATCATCGTCTCAAATGAGTCTTC
	AGAGGGTTCT

hsa-mir-	GAGGACTCCATTTGTTTTGATGAT	146
136-prec	GGATTCTTATGCTCCATCATCGTC
	TCAAATGAGTCTTC

hsa-mir-	CTTCGGTGACGGGTATTCTTGGGT	147
137-prec	GGATAATACGGATTACGTTGTTAT
	TGCTTAAGAATACGCGTAGTCGAG
	G

hsa-mir-	CCCTGGCATGGTGTGGTGGGGCAG	148
138-1-prec	CTGGTGTTGTGAATCAGGCCGTTG
	CCAATCAGAGAACGGCTACTTCAC
	AACACCAGGGCCACACCACACTAC
	AGG

hsa-mir-	CGTTGCTGCAGCTGGTGTTGTGAA	149
138-2-prec	TCAGGCCGACGAGCAGCGCATCCT
	CTTACCCGGCTATTTCACGACACC
	AGGGTTGCATCA

hsa-mir-	CAGCTGGTGTTGTGAATCAGGCCG	150
138-prec	ACGAGCAGCGCATCCTCTTACCCG
	GCTATTTCACGACACCAGGGTTG

hsa-mir-	GTGTATTCTACAGTGCACGTGTCT	151
139-prec	CCAGTGTGGCTCGGAGGCTGGAGA
	CGCGGCCCTGTTGGAGTAAC

hsa-mir-	TGTGTCTCTCTCTGTGTCCTGCCA	152
140	GTGGTTTTACCCTATGGTAGGTTA
	CGTCATGCTGTTCTACCACAGGGT
	AGAACCACGGACAGGATACCGGGG
	CACC

hsa-mir-	TCCTGCCAGTGGTTTTACCCTATG	153
140as-prec	GTAGGTTACGTCATGCTGTTCTAC
	CACAGGGTAGAACCACGGACAGGA

hsa-mir-	CCTGCCAGTGGTTTTACCCTATGG	154
140s-prec	TAGGTTACGTCATGCTGTTCTACC
	ACAGGGTAGAACCACGGACAGG

hsa-mir-	CGGCCGGCCCTGGGTCCATCTTCC	155
141-prec	AGTACAGTGTTGGATGGTCTAATT
	GTGAAGCTCCTAACACTGTCTGGT
	AAAGATGGCTCCCGGGTGGGTTC

hsa-mir-	GGGTCCATCTTCCAGTACAGTGTT	156
141-prec	GGATGGTCTAATTGTGAAGCTCCT
	AACACTGTCTGGTAAAGATGGCCC

hsa-mir-	ACCCATAAAGTAGAAAGCACTACT	157
142as-prec	AACAGCACTGGAGGGTGTAGTGTT
	TCCTACTTTATGGATG

hsa-mir-	GACAGTGCAGTCACCCATAAAGTA	158
142-prec	GAAAGCACTACTAACAGCACTGGA
	GGGTGTAGTGTTTCCTACTTTATG
	GATGAGTGTACTGTG

hsa-mir-	ACCCATAAAGTAGAAAGCACTACT	159
142s-pres	AACAGCACTGGAGGGTGTAGTGTT
	TCCTACTTTATGGATG

hsa-mir-	GCGCAGCGCCCTGTCTCCCAGCCT	160
143-prec	GAGGTGCAGTGCTGCATCTCTGGT
	CAGTTGGGAGTCTGAGATGAAGCA
	CTGTAGCTCAGGAAGAGAGAAGTT
	GTTCTGCAGC

hsa-mir-	CCTGAGGTGCAGTGCTGCATCTCT	161
143-prec	GGTCAGTTGGGAGTCTGAGATGAA
	GCACTGTAGCTCAGG

hsa-mir-	TGGGGCCCTGGCTGGGATATCATC	162
144-prec	ATATACTGTAAGTTTGCGATGAGA
	CACTACAGTATAGATGATGTACTA
	GTCCGGGCACCCCC

hsa-mir-	GGCTGGGATATCATCATATACTGT	163
144-prec	AAGTTTGCGATGAGACACTACAGT
	ATAGATGATGTACTAGTC

hsa-mir-	CACCTTGTCCTCACGGTCCAGTTT	164
145-prec	TCCCAGGAATCCCTTAGATGCTAA
	GATGGGGATTCCTGGAAATACTGT
	TCTTGAGGTCATGGTT

hsa-mir-	CTCACGGTCCAGTTTTCCCAGGAA	165
145-prec	TCCCTTAGATGCTAAGATGGGGAT
	TCCTGGAAATACTGTTCTTGAG

hsa-mir-	CCGATGTGTATCCTCAGCTTTGAG	166
146-prec	AACTGAATTCCATGGGTTGTGTCA
	GTGTCAGACCTCTGAAATTCAGTT
	CTTCAGCTGGGATATCTCTGTCAT
	CGT

hsa-mir-	AGCTTTGAGAACTGAATTCCATGG	167
146-prec	GTTGTGTCAGTGTCAGACCTGTGA
	AATTCAGTTCTTCAGCT

hsa-mir-	AATCTAAAGACAACATTTCTGCAC	168
147-prec	ACACACCAGACTATGGAAGCCAGT
	GTGTGGAAATGCTTCTGCTAGATT

hsa-mir-	GAGGCAAAGTTCTGAGACACTCCG	169
148-prec	ACTCTGAGTATGATAGAAGTCAGT
	GCACTACAGAACTTTGTCTC

hsa-mir-	GCCGGCGCCCGAGCTCTGGCTCCG	170
149-prec	TGTCTTCACTCCCGTGCTTGTCCG
	AGGAGGGAGGGAGGGACGGGGGCT
	GTGCTGGGGCAGCTGGA

hsa-mir-	GCTCTGGCTCCGTGTCTTCACTCC	171
149-prec	CGTGCTTGTCCGAGGAGGGAGGGA
	GGGAC

hsa-mir-	CTCCCCATGGCCCTGTCTCCCAAC	172
150-prec	CCTTGTACCAGTGCTGGGCTCAGA
	CCCTGGTACAGGCCTGGGGGACAG
	GGACCTGGGGAC

hsa-mir-	CCCTGTCTCCCAACCCTTGTACCA	173
150-prec	GTGCTGGGCTCAGACCCTGGTACA
	GGCCTGGGGGACAGGG

hsa-mir-	CCTGCCCTCGAGGAGCTCACAGTC	174
151-prec	TAGTATGTCTCATCCCCTACTAGA
	CTGAAGCTCCTTGAGGACAGG

hsa-mir-	TGTCCCCCCCGGCCCAGGTTCTGT	175
152-prec	GATACACTCCGACTCGGGCTCTGG
	AGCAGTCAGTGCATGACAGAACTT
	GGGCCCGGAAGGACC

hsa-mir-	GGCCCAGGTTCTGTGATACACTCC	176
152-prec	GACTCGGGCTCTGGAGCAGTCAGT
	GCATGACAGAACTTGGGCCCCGG

hsa-mir-	CTCACAGCTGCCAGTGTCATTTTT	177
153-1-prec	GTGATCTGCAGCTAGTATTCTCAC
	TCCAGTTGCATAGTCACAAAAGTG
	ATCATTGGCAGGTGTGGC

hsa-mir-	tctctctctccctcACAGCTGCCA	178
153-1-prec	GTGTCATTGTCACAAAAGTGATCA
	TTGGCAGGTGTGGCTGCTGCATG

hsa-mir-	AGCGGTGGCCAGTGTCATTTTTGT	179
153-2-prec	GATGTTGCAGCTAGTAATATGAGC
	CCAGTTGCATAGTCACAAAAGTGA
	TCATTGGAAACTGTG

hsa-mir-	CAGTGTCATTTTTGTGATGTTGCA	180
153-2-prec	GCTAGTAATATGAGCCCAGTTGCA
	TAGTCACAAAAGTGATCATTG

hsa-mir-	GTGGTACTTGAAGATAGGTTATCC	181
154-prec	GTGTTGCCTTCGCTTTATTTGTGA
	CGAATCATACACGGTTGACCTATT
	TTTCAGTACCAA

hsa-mir-	GAAGATAGGTTATCCGTGTTGCCT	182
154-prec	TCGCTTTATTTGTGACGAATCATA
	CACGGTTGACCTATTTTT

hsa-mir-	CTGTTAATGCTAATCGTGATAGGG	183
155-prec	GTTTTTGCCTCCAACTGACTCCTA
	CATATTAGCATTAACAG

hsa-mir-	CAATGTCAGCAGTGCCTTAGCAGC	184
16-2-prec	ACGTAAATATTGGCGTTAAGATTC
	TAAAATTATCTCCAGTATTAACTG
	TGCTGCTGAAGTAAGGTTGACCAT
	ACTCTACAGTTG

hsa-mir-	AGAAGGGCTATCAGGCCAGCCTTC	185
181a-prec	AGAGGACTCCAAGGAACATTCAAC
	GCTGTCGGTGAGTTTGGGATTTGA
	AAAAACCACTGACCGTTGACTGTA
	CCTTGGGGTCCTTA

hsa-mir-	TGAGTTTTGAGGTTGCTTCAGTGA	186
181b-prec	ACATTCAACGCTGTCGGTGAGTTT
	GGAATTAAAATCAAAACCATCGAC
	CGTTGATTGTACCCTATGGCTAAC
	CATCATCTACTCCA

hsa-mir-	CGGAAAATTTGCCAAGGGTTTGGG	187
181c-prec	GGAACATTCAACCTGTCGGTGAGT
	TTGGGCAGCTCAGGCAAACCATCG
	ACCGTTGAGTGGACCCTGAGGCCT
	GGAATTGCCATCCT

hsa-mir-	GAGCTGCTTGCCTCCCCCCGTTTT	188
182-as-prec	TGGCAATGGTAGAACTCACACTGG
	TGAGGTAACAGGATCCGGTGGTTC
	TAGACTTGCCAACTATGGGGCGAG
	GACTCAGCCGGCAC

hsa-mir-	TTTTTGGCAATGGTAGAACTCACA	189
182-prec	CTGGTGAGGTAACAGGATCCGGTG
	GTTCTAGACTTGCCAACTATGG

hsa-mir-	CCGCAGAGTGTGACTCCTGTTCTG	190
183-prec	TGTATGGCACTGGTAGAATTCACT
	GTGAACAGTCTCAGTCAGTGAATT
	ACCGAAGGGCCATAAACAGAGCAG
	AGACAGATCCACGA

hsa-mir-	CCAGTCACGTCCCCTTATCACTTT	191
184-prec	TCCAGCCCAGCTTTGTGACTGTAA
	GTGTTGGACGGAGAACTGATAAGG
	GTAGGTGATTGA

hsa-mir-	CCTTATCACTTTTCCAGCCCAGCT	192
184-prec	TTGTGACTGTAAGTGTTGGACGGA
	GAACTGATAAGGGTAGG

hsa-mir-	AGGGGGCGAGGGATTGGAGAGAAA	193
185-prec	GGCAGTTCCTGATGGTCCCCTCCC
	CAGGGGCTGGCTTTCCTCTGGTCC
	TTCCCTCCCA

hsa-mir-	AGGGATTGGAGAGAAAGGCAGTTC	194
185-prec	CTGATGGTCCCCTCCCCAGGGGCT
	GGCTTTCCTCTGGTCCTT

hsa-mir-	TGCTTGTAACTTTCCAAAGAATTC	195
186-prec	TCCTTTTGGGCTTTCTGGTTTTAT
	TTTAAGCCCAAAGGTGAATTTTTT
	GGGAAGTTTGAGCT

hsa-mir-	ACTTTCCAAAGAATTCTCCTTTTG	196
186-prec	GGCTTTCTGGTTTTATTTTAAGCC
	CAAAGGTGAATTTTTTGGGAAGT

hsa-mir-	GGTCGGGCTCACCATGACACAGTG	197
187-prec	TGAGACTCGGGCTACAACACAGGA
	CCCGGGGCGCTGCTCTGACCCCTC
	GTGTCTTGTGTTGCAGCCGGAGGG
	ACGCAGGTCCGCA

hsa-mir-	TGCTCCCTCTCTCACATCCCTTGC	198
188-prec	ATGGTGGAGGGTGAGCTTTCTGAA
	AACCCCTCCCACATGCAGGGTTTG
	CAGGATGGCGAGCC

hsa-mir-	TCTCACATCCCTTGCATGGTGGAG	199
188-prec	GGTGAGCTTTCTGAAAACCCCTCC
	CACATGCAGGGTTTGCAGGA

hsa-mir-	CTGTCGATTGGACCCGCCCTCCGG	200
189-prec	TGCCTACTGAGCTGATATCAGTTC
	TCATTTTACACACTGGCTCAGTTC
	AGCAGGAACAGGAGTCGAGCCCTT
	GAGCAA

hsa-mir-	CTCCGGTGCCTACTGAGCTGATAT	201
189-prec	CAGTTCTCATTTTACACACTGGCT
	CAGTTCAGCAGGAACAGGAG

hsa-mir-	TGCAGGCCTCTGTGTGATATGTTT	202
190-prec	GATATATTAGGTTGTTATTTAATC
	CAACTATATATCAAACATATTCCT
	ACAGTGTCTTGCC

hsa-mir-	CTGTGTGATATGTTTGATATATTA	203
190-prec	GGTTGTTATTTAATCCAACTATAT
	ATCAAACATATTCCTACAG

hsa-mir-	CGGCTGGACAGCGGGCAACGGAAT	204
191-prec	CCCAAAAGCAGCTGTTGTCTCCAG
	AGCATTCCAGCTGCGCTTGGATTT
	CGTCCCCTGCTCTCCTGCCT

hsa-mir-	AGCGGGCAACGGAATCCCAAAAGC	205
191-prec	AGCTGTTGTCTCCAGAGCATTCCA
	GCTGCGCTTGGATTTCGTCCCCTG
	CT

hsa-mir-	CCGAGACCGAGTGCACAGGGCTCT	206
192-2/3	GACCTATGAATTGACAGCCAGTGC
	TCTCGTCTCCCCTCTGGCTGCCAA
	TTCCATAGGTCACAGGTATGTTCG
	CCTCAATGCCAG

hsa-mir-	GCCGAGACCGAGTGCACAGGGCTC	207
192-prec	TGACCTATGAATTGACAGCCAGTG
	CTCTCGTCTCCCCTCTGGCTGCCA
	ATTCCATAGGTCACAGGTATGTTC
	GCCTCAATGCCAGC

hsa-mir-	CGAGGATGGGAGCTGAGGGCTGGG	208
193-prec	TCTTTGCGGGCGAGATGAGGGTGT
	CGGATCAACTGGCCTACAAAGTCC
	CAGTTCTCGGCCCCCG

hsa-mir-	GCTGGGTCTTTGCGGGCGAGATGA	209
193-prec	GGGTGTCGGATCAACTGGCCTACA
	AAGTCCCAGT

hsa-mir-	ATGGTGTTATCAAGTGTAACAGCA	210
194-prec	ACTCCATGTGGACTGTGTACCAAT
	TTCCAGTGGAGATGCTGTTACTTT
	TGATGGTTACCAA

hsa-mir-	GTGTAACAGCAACTCCATGTGGAC	211
194-prec	TGTGTACCAATTTCCAGTGGAGAT
	GCTGTTACTTTTGAT

hsa-mir-	AGCTTCCCTGGCTCTAGCAGCACA	212
195-prec	GAAATATTGGCACAGGGAAGCGAG
	TCTGCCAATATTGGCTGTGCTGCT
	CCAGGCAGGGTGGTG

hsa-mir-	TAGCAGCACAGAAATATTGGCACA	213
195-prec	GGGAAGCGAGTCTGCCAATATTGG
	CTGTGCTGCT

hsa-mir-	CTAGAGCTTGAATTGGAACTGCTG	214
196-1-prec	AGTGAATTAGGTAGTTTCATGTTG
	TTGGGCCTGGGTTTCTGAACACAA
	CAACATTAAACCACCCGATTCACG
	GCAGTTACTGCTCC

hsa-mir-	GTGAATTAGGTAGTTTCATGTTGT	215
196-1-prec	TGGGCCTGGGTTTCTGAACACAAC
	AACATTAAACCACCCGATTCAC

hsa-mir-	TGCTCGCTCAGCTGATCTGTGGCT	216
196-2-prec	TAGGTAGTTTCATGTTGTTGGGAT
	TGAGTTTTGAACTCGGCAACAAGA
	AACTGCCTGAGTTACATCAGTCGG
	TTTTCGTCGAGGGC

hsa-mir-	GTGAATTAGGTAGTTTCATGTTGT	217
196-prec	TGGGCCTGGGTTTCTGAACACAAC
	AACATTAAACCACCCGATTCAC

hsa-mir-	GGCTGTGCCGGGTAGAGAGGGCAG	218
197-prec	TGGGAGGTAAGAGCTCTTCACCCT
	TCACCACCTTCTCCACCCAGCATG
	GCC

hsa-mir-	TCATTGGTCCAGAGGGGAGATAGG	219
198-prec	TTCCTGTGATTTTTCCTTCTTCTC
	TATAGAATAAATGA

hsa-mir-	GCCAACCCAGTGTTCAGACTACCT	220
199a-1-prec	GTTCAGGAGGCTCTCAATGTGTAC
	AGTAGTCTGCACATTGGTTAGGC

hsa-mir-	AGGAAGCTTCTGGAGATCCTGCTC	221
199a-2-prec	CGTCGCCCCAGTGTTCAGACTACC
	TGTTCAGGACAATGCCGTTGTACA
	GTAGTCTGCACATTGGTTAGACTG
	GGCAAGGGAGAGCA

hsa-mir-	CCAGAGGACACCTCCACTCCGTCT	222
199b-prec	ACCCAGTGTTTAGACTATCTGTTC
	AGGACTCCCAAATTGTACAGTAGT
	CTGCACATTGGTTAGGCTGGGCTG
	GGTTAGACCCTCGG

hsa-mir-	GCCAACCCAGTGTTCAGACTACCT	223
199s-prec	GTTCAGGAGGCTCTCAATGTGTAC
	AGTAGTCTGCACATTGGTTAGGC

hsa-mir-	GCCGTGGCCATCTTACTGGGCAGC	224
200a-prec	ATTGGATGGAGTCAGGTCTCTAAT
	ACTGCCTGGTAATGATGACGGC

hsa-mir-	CCAGCTCGGGCAGCCGTGGCCATC	225
200b-prec	TTACTGGGCAGCATTGGATGGAGT
	CAGGTCTCTAATACTGCCTGGTAA
	TGATGACGGCGGAGCCCTGCACG

hsa-mir-	GTTCCTTTTTCCTATGCATATACT	226
202-prec	TCTTTGAGGATCTGGCCTAAAGAG
	GTATAGGGCATGGGAAGATGGAGC

hsa-mir-	GTGTTGGGGACTCGCGCGCTGGGT	227
203-prec	CCAGTGGTTCTTAACAGTTCAACA
	GTTCTGTAGCGCAATTGTGAAATG
	TTTAGGACCACTAGACCCGGCGGG
	CGCGGCGACAGCGA

hsa-mir-	GGCTACAGTCTTTCTTCATGTGAC	228
204-prec	TCGTGGACTTCCCTTTGTCATCCT
	ATGCCTGAGAATATATGAAGGAGG
	CTGGGAAGGCAAAGGGACGTTCAA
	TTGTCATCACTGGC

hsa-mir-	AAAGATCCTCAGACAATCCATGTG	229
205-prec	CTTCTCTTGTCCTTCATTCCACCG
	GAGTCTGTCTCATACCCAACCAGA
	TTTCAGTGGAGTGAAGTTCAGGAG
	GCATGGAGCTGACA

hsa-mir-	TGCTTCCCGAGGCCACATGCTTCT	230
206-prec	TTATATCCCCATATGGATTACTTT
	GCTATGGAATGTAAGGAAGTGTGT
	GGTTTCGGCAAGTG

hsa-mir-	AGGCCACATGCTTCTTTATATCCC	231
206-prec	CATATGGATTACTTTGCTATGGAA
	TGTAAGGAAGTGTGTGGTTTT

hsa-mir-	TGACGGGCGAGCTTTTGGCCCGGG	232
208-prec	TTATACCTGATGCTCACGTATAAG
	ACGAGCAAAAAGCTTGTTGGTCA

hsa-mir-	ACCCGGCAGTGCCTCCAGGCGCAG	233
210-prec	GGCAGCCCCTGCCCACCGCACACT
	GCGCTGCCCCAGACCCACTGTGCG
	TGTGACAGCGGCTGATCTGTGCCT
	GGGCAGCGCGACCC

hsa-mir-	TCACCTGGCCATGTGACTTGTGGG	234
211-prec	CTTCCCTTTGTCATCCTTCGCCTA
	GGGCTCTGAGCAGGGCAGGGACAG
	CAAAGGGGTGCTCAGTTGTCACTT
	CCCACAGCACGGAG

hsa-mir-	CGGGGCACCCCGCCCGGACAGCGC	235
212-prec	GCCGGCACCTTGGCTCTAGACTGC
	TTACTGCCCGGGCCGCCCTCAGTA
	ACAGTCTCCAGTCACGGCCACCGA
	CGCCTGGCCCCGCC

hsa-mir-	CCTGTGCAGAGATTATTTTTTAAA	236
213-prec	AGGTCACAATCAACATTCATTGCT
	GTCGGTGGGTTGAACTGTGTGGAC
	AAGCTCACTGAACAATGAATGCAA
	CTGTGGCCCCGCTT

hsa-mir-	GAGTTTTGAGGTTGCTTCAGTGAA	237
213-prec-	CATTCAACGCTGTCGGTGAGTTTG
LIM	GAATTAAAATCAAAACCATCGACC
	GTTGATTGTACCCTATGGCTAACC
	ATCATCTACTCC

hsa-mir-	GGCCTGGCTGGACAGAGTTGTCAT	238
214-prec	GTGTCTGCCTGTCTACACTTGCTG
	TGCAGAACATCCGCTCACCTGTAC
	AGCAGGCACAGACAGGCAGTCACA
	TGACAACCCAGCCT

hsa-mir-	ATCATTCAGAAATGGTATACAGGA	239
215-prec	AAATGACCTATGAATTGACAGACA
	ATATAGCTGAGTTTGTCTGTCATT
	TCTTTAGGCCAATATTCTGTATGA
	CTGTGCTACTTCAA

hsa-mir-	GATGGCTGTGAGTTGGCTTAATCT	240
216-prec	CAGCTGGCAACTGTGAGATGTTCA
	TACAATCCCTCACAGTGGTCTCTG
	GGATTATGCTAAACAGAGCAATTT
	CCTAGCCCTCACGA

hsa-mir-	AGTATAATTATTACATAGTTTTTG	241
217-prec	ATGTCGCAGATACTGCATCAGGAA
	CTGATTGGATAAGAATCAGTCACC
	ATCAGTTCCTAATGCATTGCCTTC
	AGCATCTAAACAAG

hsa-mir-	GTGATAATGTAGCGAGATTTTCTG	242
218-1-prec	TTGTGCTTGATCTAACCATGTGGT
	TGCGAGGTATGAGTAAAACATGGT
	TCCGTCAAGCACCATGGAACGTCA
	CGCAGCTTTCTACA

hsa-mir-	GACCAGTCGCTGCGGGGCTTTCCT	243
218-2-prec	TTGTGCTTGATCTAACCATGTGGT
	GGAACGATGGAAACGGAACATGGT
	TCTGTCAAGCACCGCGGAAAGCAC
	CGTGCTCTCCTGCA

hsa-mir-	CCGCCCCGGGCCGCGGCTCCTGAT	244
219-prec	TGTCCAAACGCAATTCTCGAGTCT
	ATGGCTCCGGCCGAGAGTTGAGTC
	TGGACGTCCCGAGCCGCCGCCCCC
	AAACCTCGAGCGGG

hsa-mir-	GACAGTGTGGCATTGTAGGGCTCC	245
220-prec	ACACCGTATCTGACACTTTGGGCG
	AGGGCACCATGCTGAAGGTGTTCA
	TGATGCGGTCTGGGAACTCCTCAC
	GGATCTTACTGATG

hsa-mir-	TGAACATCCAGGTCTGGGGCATGA	246
221-prec	ACCTGGCATACAATGTAGATTTCT
	GTGTTCGTTAGGCAACAGCTACAT
	TGTCTGCTGGGTTTCAGGCTACCT
	GGAAACATGTTCTC

hsa-mir-	GCTGCTGGAAGGTGTAGGTACCCT	247
222-prec	CAATGGCTCAGTAGCCAGTGTAGA
	TCCTGTCTTTCGTAATCAGCAGCT
	ACATCTGGCTACTGGGTCTCTGAT
	GGCATCTTCTAGCT

hsa-mir-	CCTGGCCTCCTGCAGTGCCACGCT	248
223-prec	CCGTGTATTTGACAAGCTGAGTTG
	GACACTCCATGTGGTAGAGTGTCA
	GTTTGTCAAATACCCCAAGTGCGG
	CACATGCTTACCAG

hsa-mir-	GGGCTTTCAAGTCACTAGTGGTTC	249
224-prec	CGTTTAGTAGATGATTGTGCATTG
	TTTCAAAATGGTGCCCTAGTGACT
	ACAAAGCCC

hsA-mir-	CTTCTGGAAGCTGGTTTCACATGG	250
29b-1 =	TGGCTTAGATTTTTCCATCTTTGT
102-prec1	ATCTAGCACCATTTGAAATCAGTG
	TTTTAGGAG

hsA-mir-	CTTCAGGAAGCTGGTTTCATATGG	251
29b-2 =	TGGTTTAGATTTAAATAGTGATTG
102prec7.1 =	TCTAGCACCATTTGAAATCAGTGT
7.2	TCTTGGGGG

hsA-mir-	CTTCAGGAAGCTGGTTTCATATGG	252
29b-3 =	TGGTTTAGATTTAAATAGTGATTG
102prec7.1 =	TCTAGCACCATTTGAAATCAGTGT
7.2	TCTTGGGGG

hsa-mir-	GTGAGCGACTGTAAACATCCTCGA	253
30* = mir-	CTGGAAGCTGTGAAGCCACAGATG
097-prec-6	GGCTTTCAGTCGGATGTTTGCAGC
	TGCCTACT

mir-033b	ACCAAGTTTCAGTTCATGTAAACA	254
	TCCTACACTCAGCTGTAATACATG
	GATTGGCTGGGAGGTGGATGTTTA
	CTTCAGCTGACTTGGA

mir-101-	TGCCCTGGCTCAGTTATCACAGTG	255
precursor-9 =	CTGATGCTGTCTATTCTAAAGGTA
mir-101-3	CAGTACTGTGATAACTGAAGGATG
	GCA

mir-108-1-	ACACTGCAAGAACAATAAGGATTT	256
small	TTAGGGGCATTATGACTGAGTCAG
	AAAACACAGCTGCCCCTGAAAGTC
	CCTCATTTTTCTTGCTGT

mir-108-2-	ACTGCAAGAGCAATAAGGATTTTT	257
small	AGGGGCATTATGATAGTGGAATGG
	AAACACATCTGCCCCCAAAAGTCC
	CTCATTTT

mir-123-	CGCTGGCGACGGGACATTATTACT	258
prec = mir-	TTTGGTACGCGCTGTGACACTTCA
126-prec	AACTCGTACCGTGAGTAATAATGC
	GCCGTCCACGGCA

mir-123-	ACATTATTACTTTTGGTACGCGCT	259
prec = mir-	GTGACACTTCAAACTCGTACCGTG
126-prec	AGTAATAATGCGC

mir-129-	TGGATCTTTTTGCGGTCTGGGCTT	260
1-prec	GCTGTTCCTCTCAACAGTAGTCAG
	GAAGCCCTTACCCCAAAAAGTATC
	TA

mir-129-	TGCCCTTCGCGAATCTTTTTGCGG	261
small-2-129b?	TCTGGGCTTGCTGTACATAACTCA
	ATAGCCGGAAGCCCTTACCCCAAA
	AAGCATTTGCGGAGGGCG

mir-133b-	GCCCCCTGCTCTGGCTGGTCAAAC	262
small	GGAACCAAGTCCGTCTTCCTGAGA
	GGTTTGGTCCCCTTCAACCAGCTA
	CAGCAGGG

mir-135-	AGATAAATTCACTCTAGTGCTTTA	263
small-2	TGGCTTTTTATTCCTATGTGATAG
	TAATAAAGTCTCATGTAGGGATGG
	AAGCCATGAAATACATTGTGAAAA
	ATCA

mir-148b-	AAGCACGATTAGCATTTGAGGTGA	264
small	AGTTCTGTTATACACTCAGGCTGT
	GGCTCTCTGAAAGTCAGTGCAT

mir-151-	CCTGTCCTCAAGGAGCTTCAGTCT	265
prec	AGTAGGGGATGAGACATACTAGAC
	TGTGAGCTCCTCGAGGGCAGG

mir-155-	CTGTTAATGCTAATCGTGATAGGG	266
prec(BIC)	GTTTTTGCCTCCAACTGACTCCTA
	CATATTAGCATTAACAG

mir-156 =	CCTAACACTGTCTGGTAAAGATGG	267
mir-157 =	CTCCCGGGTGGGTTCTCTCGGCAG
overlap	TAACCTTCAGGGAGCCCTGAAGAC
mir-141	CATGGAGGAC

mir-158-	GCCGAGACCGAGTGCACAGGGCTC	268
small =	TGACCTATGAATTGACAGCCAGTG
mir-192	CTCTCGTCTCCCCTCTGGCTGCCA
	ATTCCATAGGTCACAGGTATGTTC
	GCCTCAATGCCAGC

mir-159-	TCCCGCCCCCTGTAACAGCAACTC	269
1-small	CATGTGGAAGTGCCCACTGGTTCC
	AGTGGGGCTGCTGTTATCTGGGGC
	GAGGGCCA

mir-161-	AAAGCTGGGTTGAGAGGGCGAAAA	270
small	AGGATGAGGTGACTGGTCTGGGCT
	ACGCTATGCTGCGGCGCTCGGG

mir-163-	CATTGGCCTCCTAAGCCAGGGATT	271
1b-small	GTGGGTTCGAGTCCCACCCGGGGT
	AAAGAAAGGCCGAATT

mir-163-	CCTAAGCCAGGGATTGTGGGTTCG	272
3-small	AGTCCCACCTGGGGTAGAGGTGAA
	AGTTCCTTTTACGGAATTTTTT

mir-175-	GGGCTTTCAAGTCACTAGTGGTTC	273
small =	CGTTTAGTAGATGATTGTGCATTG
mir-224	TTTCAAAATGGTGCCCTAGTGACT
	ACAAAGCCC

mir-177-	ACGCAAGTGTCCTAAGGTGAGCTC	274
small	AGGGAGCACAGAAACCTCCAGTGG
	AACAGAAGGGCAAAAGCTCATT

mir-180-	CATGTGTCACTTTCAGGTGGAGTT	275
small	TCAAGAGTCCCTTCCTGGTTCACC
	GTCTCCTTTGCTCTTCCACAAC

mir-187-	GGTCGGGCTCACCATGACACAGTG	276
prec	TGAGACTCGGGCTACAACACAGGA
	CCCGGGGCGCTGCTCTGACCCCTC
	GTGTCTTGTGTTGCAGCCGGAGGG
	ACGCAGGTCCGCA

mir-188-	TGCTCCCTCTCTCACATCCCTTGC	277
prec	ATGGTGGAGGGTGAGCTTTCTGAA
	AACCCCTCCCACATGCAGGGTTTG
	CAGGATGGCGAGCC

mir-190-	TGCAGGCCTCTGTGTGATATGTTT	278
prec	GATATATTAGGTTGTTATTTAATC
	CAACTATATATCAAACATATTCCT
	ACAGTGTCTTGCC

mir-197-2	GTGCATGTGTATGTATGTGTGCAT	279
	GTGCATGTGTATGTGTATGAGTGC
	ATGCGTGTGTGC

mir-197-	GGCTGTGCCGGGTAGAGAGGGCAG	280
prec	TGGGAGGTAAGAGCTCTTCACCCT
	TCACCACCTTCTCCACCCAGCATG
	GCC

mir-202-	GTTCCTTTTTCCTATGCATATACT	281
prec	TCTTTGAGGATCTGGCCTAAAGAG
	GTATAGGGCATGGGAAGATGGAGC

mir-294-1	CAATCTTCCTTTATCATGGTATTG	282
(chr16)	ATTTTTCAGTGCTTCCCTTTTGTG
	TGAGAGAAGATA

mir-hesl	ATGGAGCTGCTCACCCTGTGGGCC	283
	TCAAATGTGGAGGAACTATTCTGA
	TGTCCAAGTGGAAAGTGCTGCGAC
	ATTTGAGCGTCACCGGTGACGCCC
	ATATCA

mir-hes2	GCATCCCCTCAGCCTGTGGCACTC	284
	AAACTGTGGGGGCACTTTCTGCTC
	TCTGGTGAAAGTGCCGCCATCTTT
	TGAGTGTTACCGCTTGAGAAGACT
	CAACC

mir-hes3	CGAGGAGCTCATACTGGGATACTC	285
	AAAATGGGGGCGCTTTCCTTTTTG
	TCTGTTACTGGGAAGTGCTTCGAT
	TTTGGGGTGTCCCTGTTTGAGTAG
	GGCATC

hsa-mir-	CTTCAGGAAGCTGGTTTCATATGG	286
29b-1	TGGTTTAGATTTAAATAGTGATTG
	TCTAGCACCATTTGAAATCAGTGT
	TCTTGGGGG

*An underlined sequence within a precursor sequence represents a processed miR transcript. All sequences are human.

The level of at least one miR gene product can be measured in cells of a biological sample obtained from the subject. For example, a tissue sample can be removed from a subject suspected of having breast cancer associated with by conventional biopsy techniques. In another example, a blood sample can be removed from the subject, and white blood cells can be isolated for DNA extraction by standard techniques. The blood or tissue sample is preferably obtained from the subject prior to initiation of radiotherapy, chemotherapy or other therapeutic treatment. A corresponding control tissue or blood sample can be obtained from unaffected tissues of the subject, from a normal human individual or population of normal individuals, or from cultured cells corresponding to the majority of cells in the subject's sample. The control tissue or blood sample is then processed along with the sample from the subject, so that the levels of miR gene product produced from a given miR gene in cells from the subject's sample can be compared to the corresponding miR gene product levels from cells of the control sample.
An alteration (i.e., an increase or decrease) in the level of a miR gene product in the sample obtained from the subject, relative to the level of a corresponding miR gene product in a control sample, is indicative of the presence of breast cancer in the subject. In one embodiment, the level of the at least one miR gene product in the test sample is greater than the level of the corresponding miR gene product in the control sample (i.e., expression of the miR gene product is “up-regulated”). As used herein, expression of a miR gene product is “up-regulated” when the amount of miR gene product in a cell or tissue sample from a subject is greater than the amount the same gene product in a control cell or tissue sample. In another embodiment, the level of the at least one miR gene product in the test sample is less than the level of the corresponding miR gene product in the control sample (i.e., expression of the miR gene product is “down-regulated”). As used herein, expression of a miR gene is “down-regulated” when the amount of miR gene product produced from that gene in a cell or tissue sample from a subject is less than the amount produced from the same gene in a control cell or tissue sample. The relative miR gene expression in the control and normal samples can be determined with respect to one or more RNA expression standards. The standards can comprise, for example, a zero miR gene expression level, the miR gene expression level in a standard cell line, or the average level of miR gene expression previously obtained for a population of normal human controls.
The level of a miR gene product in a sample can be measured using any technique that is suitable for detecting RNA expression levels in a biological sample. Suitable techniques for determining RNA expression levels in cells from a biological sample (for example, Northern blot analysis, RT-PCR, in situ hybridization) are well known to those of skill in the art. In a particular embodiment, the level of at least one miR gene product is detected using Northern blot analysis. For example, total cellular RNA can be purified from cells by homogenization in the presence of nucleic acid extraction buffer, followed by centrifugation. Nucleic acids are precipitated, and DNA is removed by treatment with DNase and precipitation. The RNA molecules are then separated by gel electrophoresis on agarose gels according to standard techniques, and transferred to nitrocellulose filters. The RNA is then immobilized on the filters by heating. Detection and quantification of specific RNA is accomplished using appropriately labeled DNA or RNA probes complementary to the RNA in question. See, for example, Molecular Cloning: A Laboratory Manual, J. Sambrook et al., eds., 2nd edition, Cold Spring Harbor Laboratory Press, 1989, Chapter 7, the entire disclosure of which is incorporated by reference.
Suitable probes for Northern blot hybridization of a given miR gene product can be produced from the nucleic acid sequences provided in Table 1. Methods for preparation of labeled DNA and RNA probes, and the conditions for hybridization thereof to target nucleotide sequences, are described in Molecular Cloning: A Laboratory Manual, J. Sambrook et al., eds., 2nd edition, Cold Spring Harbor Laboratory Press, 1989, Chapters 10 and 11, the disclosures of which are incorporated herein by reference.
For example, the nucleic acid probe can be labeled with, for example, a radionuclide, such as ³H, ³²P, ³³P, ¹⁴C, or ³⁵S; a heavy metal; or a ligand capable of functioning as a specific binding pair member for a labeled ligand (for example, biotin, avidin or an antibody), a fluorescent molecule, a chemiluminescent molecule, an enzyme or the like.
Probes can be labeled to high specific activity by either the nick translation method of Rigby et al. (1977), J. Mol. Biol. 113:237-251 or by the random priming method of Fienberg et al. (1983), Anal. Biochem. 132:6-13, the entire disclosures of which are incorporated herein by reference. The latter is the method of choice for synthesizing ³²P-labeled probes of high specific activity from single-stranded DNA or from RNA templates. For example, by replacing preexisting nucleotides with highly radioactive nucleotides according to the nick translation method, it is possible to prepare ³²P-labeled nucleic acid probes with a specific activity well in excess of 10⁸cpm/microgram. Autoradiographic detection of hybridization can then be performed by exposing hybridized filters to photographic film. Densitometric scanning of the photographic films exposed by the hybridized filters provides an accurate measurement of miR gene transcript levels. Using another approach, miR gene transcript levels can be quantified by computerized imaging systems, such the Molecular Dynamics 400-B 2D Phosphorimager available from Amersham Biosciences, Piscataway, N.J.
Where radionuclide labeling of DNA or RNA probes is not practical, the random-primer method can be used to incorporate an analogue, for example, the dTTP analogue 5-(N—(N-biotinyl-epsilon-aminocaproyl)-3-aminoallyl)deoxyuridine triphosphate, into the probe molecule. The biotinylated probe oligonucleotide can be detected by reaction with biotin-binding proteins, such as avidin, streptavidin, and antibodies (for example, anti-biotin antibodies) coupled to fluorescent dyes or enzymes that produce color reactions.
In addition to Northern and other RNA hybridization techniques, determining the levels of RNA transcripts can be accomplished using the technique of in situ hybridization. This technique requires fewer cells than the Northern blotting technique, and involves depositing whole cells onto a microscope cover slip and probing the nucleic acid content of the cell with a solution containing radioactive or otherwise labeled nucleic acid (for example, cDNA or RNA) probes. This technique is particularly well-suited for analyzing tissue biopsy samples from subjects. The practice of the in situ hybridization technique is described in more detail in U.S. Pat. No. 5,427,916, the entire disclosure of which is incorporated herein by reference. Suitable probes for in situ hybridization of a given miR gene product can be produced from the nucleic acid sequences provided in Table 1, as described above.
The relative number of miR gene transcripts in cells can also be determined by reverse transcription of miR gene transcripts, followed by amplification of the reverse-transcribed transcripts by polymerase chain reaction (RT-PCR). The levels of miR gene transcripts can be quantified in comparison with an internal standard, for example, the level of mRNA from a “housekeeping” gene present in the same sample. A suitable “housekeeping” gene for use as an internal standard includes, for example, myosin or glyceraldehyde-3-phosphate dehydrogenase (G3PDH). The methods for quantitative RT-PCR and variations thereof are within the skill in the art.
In some instances, it may be desirable to simultaneously determine the expression level of a plurality of different miR gene products in a sample. In other instances, it may be desirable to determine the expression level of the transcripts of all known miR genes correlated with a cancer. Assessing cancer-specific expression levels for hundreds of miR genes is time consuming and requires a large amount of total RNA (at least 20 μg for each Northern blot) and autoradiographic techniques that require radioactive isotopes.
To overcome these limitations, an oligolibrary, in microchip format (i.e., a microarray), may be constructed containing a set of probe oligodeoxynucleotides that are specific for a set of miR genes. Using such a microarray, the expression level of multiple microRNAs in a biological sample can be determined by reverse transcribing the RNAs to generate a set of target oligodeoxynucleotides, and hybridizing them to probe oligodeoxynucleotides on the microarray to generate a hybridization, or expression, profile. The hybridization profile of the test sample can then be compared to that of a control sample to determine which microRNAs have an altered expression level in breast cancer cells. As used herein, “probe oligonucleotide” or “probe oligodeoxynucleotide” refers to an oligonucleotide that is capable of hybridizing to a target oligonucleotide. “Target oligonucleotide” or “target oligodeoxynucleotide” refers to a molecule to be detected (for example, via hybridization). By “miR-specific probe oligonucleotide” or “probe oligonucleotide specific for a miR” is meant a probe oligonucleotide that has a sequence selected to hybridize to a specific miR gene product, or to a reverse transcript of the specific miR gene product.
An “expression profile” or “hybridization profile” of a particular sample is essentially a fingerprint of the state of the sample; while two states may have any particular gene similarly expressed, the evaluation of a number of genes simultaneously allows the generation of a gene expression profile that is unique to the state of the cell. That is, normal tissue may be distinguished from breast cancer tissue, and within breast cancer tissue, different prognosis states (good or poor long term survival prospects, for example) may be determined. By comparing expression profiles of breast cancer tissue in different states, information regarding which genes are important (including both up- and down-regulation of genes) in each of these states is obtained. The identification of sequences that are differentially expressed in breast cancer tissue or normal breast tissue, as well as differential expression resulting in different prognostic outcomes, allows the use of this information in a number of ways. For example, a particular treatment regime may be evaluated (for example, to determine whether a chemotherapeutic drug act to improve the long-term prognosis in a particular patient). Similarly, diagnosis may be done or confirmed by comparing patient samples with the known expression profiles. Furthermore, these gene expression profiles (or individual genes) allow screening of drug candidates that suppress the breast cancer expression profile or convert a poor prognosis profile to a better prognosis profile.
Accordingly, the invention provides methods of diagnosing whether a subject has, or is at risk for developing, breast cancer, comprising reverse transcribing RNA from a test sample obtained from the subject to provide a set of target oligo-deoxynucleotides, hybridizing the target oligo-deoxynucleotides to a microarray comprising miRNA-specific probe oligonucleotides to provide a hybridization profile for the test sample, and comparing the test sample hybridization profile to a hybridization profile generated from a control sample, wherein an alteration in the signal of at least one miRNA is indicative of the subject either having, or being at risk for developing, breast cancer. In one embodiment, the microarray comprises miRNA-specific probe oligonucleotides for a substantial portion of the human miRNome. In a particular embodiment, the microarray comprises miRNA-specific probe oligo-nucleotides for one or more miRNAs selected from the group consisting of miR-125b, miR-145, miR-21, miR-155, miR-10b, miR-009-1 (miR131-1), miR-34 (miR-170), miR-102 (miR-29b), miR-123 (miR-126), miR-140-as, miR-125a, miR-125b-1, miR-125b-2, miR-194, miR-204, miR-213, let-7a-2, let-7a-3, let-7d (let-7d-v1), let-7f-2, let-7i (let-7d-v2), miR-101-1, miR-122a, miR-128b, miR-136, miR-143, miR-149, miR-191, miR-196-1, miR-196-2, miR-202, miR-203, miR-206, miR-210 and combinations thereof. In a further embodiment, the at least one miR gene product is selected from the group consisting of miR-125b, miR-145, miR-21, miR-155, miR-10b and combinations thereof.
The microarray can be prepared from gene-specific oligonucleotide probes generated from known miRNA sequences. The array may contain two different oligonucleotide probes for each miRNA, one containing the active, mature sequence and the other being specific for the precursor of the miRNA. The array may also contain controls, such as one or more mouse sequences differing from human orthologs by only a few bases, which can serve as controls for hybridization stringency conditions. tRNAs from both species may also be printed on the microchip, providing an internal, relatively stable, positive control for specific hybridization. One or more appropriate controls for non-specific hybridization may also be included on the microchip. For this purpose, sequences are selected based upon the absence of any homology with any known miRNAs.
The microarray may be fabricated using techniques known in the art. For example, probe oligonucleotides of an appropriate length, for example, 40 nucleotides, are 5′-amine modified at position C6 and printed using commercially available microarray systems, for example, the GeneMachine OmniGrid™ 100 Microarrayer and Amersham CodeLink™ activated slides. Labeled cDNA oligomer corresponding to the target RNAs is prepared by reverse transcribing the target RNA with labeled primer. Following first strand synthesis, the RNA/DNA hybrids are denatured to degrade the RNA templates. The labeled target cDNAs thus prepared are then hybridized to the microarray chip under hybridizing conditions, for example, 6×SSPE/30% formamide at 25° C. for 18 hours, followed by washing in 0.75×TNT at 37° C. for 40 minutes. At positions on the array where the immobilized probe DNA recognizes a complementary target cDNA in the sample, hybridization occurs. The labeled target cDNA marks the exact position on the array where binding occurs, allowing automatic detection and quantification. The output consists of a list of hybridization events, indicating the relative abundance of specific cDNA sequences, and therefore the relative abundance of the corresponding complementary miRs, in the patient sample. According to one embodiment, the labeled cDNA oligomer is a biotin-labeled cDNA, prepared from a biotin-labeled primer. The microarray is then processed by direct detection of the biotin-containing transcripts using, for example, Streptavidin-Alexa647 conjugate, and scanned utilizing conventional scanning methods. Image intensities of each spot on the array are proportional to the abundance of the corresponding miR in the patient sample.
The use of the array has several advantages for miRNA expression detection. First, the global expression of several hundred genes can be identified in the same sample at one time point. Second, through careful design of the oligonucleotide probes, expression of both mature and precursor molecules can be identified. Third, in comparison with Northern blot analysis, the chip requires a small amount of RNA, and provides reproducible results using 2.5 μg of total RNA. The relatively limited number of miRNAs (a few hundred per species) allows the construction of a common microarray for several species, with distinct oligonucleotide probes for each. Such a tool would allow for analysis of trans-species expression for each known miR under various conditions.
In addition to use for quantitative expression level assays of specific miRs, a microchip containing miRNA-specific probe oligonucleotides corresponding to a substantial portion of the miRNome, preferably the entire miRNome, may be employed to carry out miR gene expression profiling, for analysis of miR expression patterns. Distinct miR signatures can be associated with established disease markers, or directly with a disease state.
According to the expression profiling methods described herein, total RNA from a sample from a subject suspected of having a cancer (such as breast cancer) is quantitatively reverse transcribed to provide a set of labeled target oligodeoxynucleotides complementary to the RNA in the sample. The target oligodeoxynucleotides are then hybridized to a microarray comprising miRNA-specific probe oligonucleotides to provide a hybridization profile for the sample. The result is a hybridization profile for the sample representing the expression pattern of miRNA in the sample. The hybridization profile comprises the signal from the binding of the target oligodeoxynucleotides from the sample to the miRNA-specific probe oligonucleotides in the microarray. The profile may be recorded as the presence or absence of binding (signal vs. zero signal). More preferably, the profile recorded includes the intensity of the signal from each hybridization. The profile is compared to the hybridization profile generated from a normal, noncancerous, control sample. An alteration in the signal is indicative of the presence of the cancer in the subject.
Other techniques for measuring miR gene expression are also within the skill in the art, and include various techniques for measuring rates of RNA transcription and degradation.
The invention also provides methods of diagnosing a breast cancer associated with one or more prognostic markers, comprising measuring the level of at least one miR gene product in a breast cancer test sample from a subject and comparing the level of the at least one miR gene product in the breast cancer test sample to the level of a corresponding miR gene product in a control sample. An alteration (for example, an increase, a decrease) in the signal of at least one miRNA in the test sample relative to the control sample is indicative of the subject either having, or being at risk for developing, breast cancer associated with the one or more prognostic markers.
The breast cancer can be associated with one or more prognostic markers or features, including, a marker associated with an adverse (negative) prognosis, or a marker associated with a good (positive) prognosis. In certain embodiments, the breast cancer that is diagnosed using the methods described herein is associated with one or more adverse prognostic features selected from the group consisting of estrogen receptor expression, progesterone receptor expression, positive lymph node metastasis, high proliferative index, detectable p53 expression, advanced tumor stage, and high vascular invasion. Particular microRNAs whose expression is altered in breast cancer cells associated with each of these prognostic markers are described herein (see, for example, Example 3 and FIG. 4). In one embodiment, the level of the at least one miR gene product is measured by reverse transcribing RNA from a test sample obtained from the subject to provide a set of target oligodeoxynucleotides, hybridizing the target oligodeoxynucleotides to a microarray that comprises miRNA-specific probe oligonucleotides to provide a hybridization profile for the test sample, and comparing the test sample hybridization profile to a hybridization profile generated from a control sample.
Without wishing to be bound by any one theory, it is believed that alterations in the level of one or more miR gene products in cells can result in the deregulation of one or more intended targets for these miRs, which can lead to the formation of breast cancer. Therefore, altering the level of the miR gene product (for example, by decreasing the level of a miR that is up-regulated in breast cancer cells and/or by increasing the level of a miR that is down-regulated in cancer cells) may successfully treat the breast cancer. Examples of putative gene targets for miRNAs that are deregulated in breast cancer tissues are described herein (see, for example, Example 2 and Table 4).
Accordingly, the present invention encompasses methods of treating breast cancer in a subject, wherein at least one miR gene product is de-regulated (for example, down-regulated or up-regulated) in the cancer cells of the subject. When the at least one isolated miR gene product is down-regulated in the breast cancer cells, the method comprises administering an effective amount of the at least one isolated miR gene product, provided that the miR gene is not miR15 or miR16, such that proliferation of cancer cells in the subject is inhibited. When the at least one isolated miR gene product is up-regulated in the cancer cells, the method comprises administering to the subject an effective amount of at least one compound for inhibiting expression of the at least one miR gene, referred to herein as miR gene expression inhibition compounds, such that proliferation of breast cancer cells is inhibited.
The terms “treat”, “treating” and “treatment”, as used herein, refer to ameliorating symptoms associated with a disease or condition, for example, breast cancer, including preventing or delaying the onset of the disease symptoms, and/or lessening the severity or frequency of symptoms of the disease or condition. The terms “subject” and “individual” are defined herein to include animals, such as mammals, including but not limited to, primates, cows, sheep, goats, horses, dogs, cats, rabbits, guinea pigs, rats, mice or other bovine, ovine, equine, canine, feline, rodent, or murine species. In a preferred embodiment, the animal is a human.
As used herein, an “effective amount” of an isolated miR gene product is an amount sufficient to inhibit proliferation of a cancer cell in a subject suffering from breast cancer. One skilled in the art can readily determine an effective amount of an miR gene product to be administered to a given subject, by taking into account factors, such as the size and weight of the subject; the extent of disease penetration; the age, health and sex of the subject; the route of administration; and whether the administration is regional or systemic.
For example, an effective amount of an isolated miR gene product can be based on the approximate weight of a tumor mass to be treated. The approximate weight of a tumor mass can be determined by calculating the approximate volume of the mass, wherein one cubic centimeter of volume is roughly equivalent to one gram. An effective amount of the isolated miR gene product based on the weight of a tumor mass can be in the range of about 10-500 micrograms/gram of tumor mass. In certain embodiments, the tumor mass can be at least about 10 micrograms/gram of tumor mass, at least about 60 micrograms/gram of tumor mass or at least about 100 micrograms/gram of tumor mass.
An effective amount of an isolated miR gene product can also be based on the approximate or estimated body weight of a subject to be treated. Preferably, such effective amounts are administered parenterally or enterally, as described herein. For example, an effective amount of the isolated miR gene product is administered to a subject can range from about 5-3000 micrograms/kg of body weight, from about 700-1000 micrograms/kg of body weight, or greater than about 1000 micrograms/kg of body weight.
One skilled in the art can also readily determine an appropriate dosage regimen for the administration of an isolated miR gene product to a given subject. For example, a miR gene product can be administered to the subject once (for example, as a single injection or deposition). Alternatively, a miR gene product can be administered once or twice daily to a subject for a period of from about three to about twenty-eight days, more particularly from about seven to about ten days. In a particular dosage regimen, a miR gene product is administered once a day for seven days. Where a dosage regimen comprises multiple administrations, it is understood that the effective amount of the miR gene product administered to the subject can comprise the total amount of gene product administered over the entire dosage regimen.
As used herein, an “isolated” miR gene product is one which is synthesized, or altered or removed from the natural state through human intervention. For example, a synthetic miR gene product, or a miR gene product partially or completely separated from the coexisting materials of its natural state, is considered to be “isolated.” An isolated miR gene product can exist in substantially-purified form, or can exist in a cell into which the miR gene product has been delivered. Thus, a miR gene product which is deliberately delivered to, or expressed in, a cell is considered an “isolated” miR gene product. A miR gene product produced inside a cell from a miR precursor molecule is also considered to be “isolated” molecule.
Isolated miR gene products can be obtained using a number of standard techniques. For example, the miR gene products can be chemically synthesized or recombinantly produced using methods known in the art. In one embodiment, miR gene products are chemically synthesized using appropriately protected ribonucleoside phosphoramidites and a conventional DNA/RNA synthesizer. Commercial suppliers of synthetic RNA molecules or synthesis reagents include, for example, Proligo (Hamburg, Germany), Dharmacon Research (Lafayette, Colo., U.S.A.), Pierce Chemical (part of Perbio Science, Rockford, Ill., U.S.A.), Glen Research (Sterling, Va., U.S.A.), ChemGenes (Ashland, Mass., U.S.A.) and Cruachem (Glasgow, UK).
Alternatively, the miR gene products can be expressed from recombinant circular or linear DNA plasmids using any suitable promoter. Suitable promoters for expressing RNA from a plasmid include, for example, the U6 or H1 RNA pol III promoter sequences, or the cytomegalovirus promoters. Selection of other suitable promoters is within the skill in the art. The recombinant plasmids of the invention can also comprise inducible or regulatable promoters for expression of the miR gene products in cancer cells.
The miR gene products that are expressed from recombinant plasmids can be isolated from cultured cell expression systems by standard techniques. The miR gene products which are expressed from recombinant plasmids can also be delivered to, and expressed directly in, the cancer cells. The use of recombinant plasmids to deliver the miR gene products to cancer cells is discussed in more detail below.
The miR gene products can be expressed from a separate recombinant plasmid, or they can be expressed from the same recombinant plasmid. In one embodiment, the miR gene products are expressed as RNA precursor molecules from a single plasmid, and the precursor molecules are processed into the functional miR gene product by a suitable processing system, including, but not limited to, processing systems extant within a cancer cell. Other suitable processing systems include, for example, the in vitro Drosophila cell lysate system (for example, as described in U.S. Published Patent Application No. 2002/0086356 to Tuschl et al., the entire disclosure of which are incorporated herein by reference) and the E. coli RNAse III system (for example, as described in U.S. Published Patent Application No. 2004/0014113 to Yang et al., the entire disclosure of which are incorporated herein by reference).
Selection of plasmids suitable for expressing the miR gene products, methods for inserting nucleic acid sequences into the plasmid to express the gene products, and methods of delivering the recombinant plasmid to the cells of interest are within the skill in the art. See, for example, Zeng et al. (2002), Molecular Cell 9:1327-1333; Tuschl (2002), Nat. Biotechnol, 20:446-448; Brummelkamp et al. (2002), Science 296:550-553; Miyagishi et al. (2002), Nat. Biotechnol. 20:497-500; Paddison et al. (2002), Genes Dev. 16:948-958; Lee et al. (2002), Nat. Biotechnol. 20:500-505; and Paul et al. (2002), Nat. Biotechnol. 20:505-508, the entire disclosures of which are incorporated herein by reference.
In one embodiment, a plasmid expressing the miR gene products comprises a sequence encoding a miR precursor RNA under the control of the CMV intermediate-early promoter. As used herein, “under the control” of a promoter means that the nucleic acid sequences encoding the miR gene product are located 3′ of the promoter, so that the promoter can initiate transcription of the miR gene product coding sequences.
The miR gene products can also be expressed from recombinant viral vectors. It is contemplated that the miR gene products can be expressed from two separate recombinant viral vectors, or from the same viral vector. The RNA expressed from the recombinant viral vectors can either be isolated from cultured cell expression systems by standard techniques, or can be expressed directly in cancer cells. The use of recombinant viral vectors to deliver the miR gene products to cancer cells is discussed in more detail below.
The recombinant viral vectors of the invention comprise sequences encoding the miR gene products and any suitable promoter for expressing the RNA sequences. Suitable promoters include, for example, the U6 or H1 RNA pol III promoter sequences, or the cytomegalovirus promoters. Selection of other suitable promoters is within the skill in the art. The recombinant viral vectors of the invention can also comprise inducible or regulatable promoters for expression of the miR gene products in a cancer cell.
Any viral vector capable of accepting the coding sequences for the miR gene products can be used; for example, vectors derived from adenovirus (AV); adeno-associated virus (AAV); retroviruses (for example, lentiviruses (LV), Rhabdoviruses, murine leukemia virus); herpes virus, and the like. The tropism of the viral vectors can be modified by pseudotyping the vectors with envelope proteins or other surface antigens from other viruses, or by substituting different viral capsid proteins, as appropriate.
For example, lentiviral vectors of the invention can be pseudotyped with surface proteins from vesicular stomatitis virus (VSV), rabies, Ebola, Mokola, and the like. AAV vectors of the invention can be made to target different cells by engineering the vectors to express different capsid protein serotypes. For example, an AAV vector expressing a serotype 2 capsid on a serotype 2 genome is called AAV 2/2. This serotype 2 capsid gene in the AAV 2/2 vector can be replaced by a serotype 5 capsid gene to produce an AAV 2/5 vector. Techniques for constructing AAV vectors that express different capsid protein serotypes are within the skill in the art; see, for example, Rabinowitz, J. E., et al. (2002), J. Virol. 76:791-801, the entire disclosure of which is incorporated herein by reference.
Selection of recombinant viral vectors suitable for use in the invention, methods for inserting nucleic acid sequences for expressing RNA into the vector, methods of delivering the viral vector to the cells of interest, and recovery of the expressed RNA products are within the skill in the art. See, for example, Dornburg (1995), Gene Therap. 2:301-310; Eglitis (1988), Biotechniques 6:608-614; Miller (1990), Hum. Gene Therap. 1:5-14; and Anderson (1998), Nature 392:25-30, the entire disclosures of which are incorporated herein by reference.
Particularly suitable viral vectors are those derived from AV and AAV. A suitable AV vector for expressing the miR gene products, a method for constructing the recombinant AV vector, and a method for delivering the vector into target cells, are described in Xia et al. (2002), Nat. Biotech. 20:1006-1010, the entire disclosure of which is incorporated herein by reference. Suitable AAV vectors for expressing the miR gene products, methods for constructing the recombinant AAV vector, and methods for delivering the vectors into target cells are described in Samulski et al. (1987), J. Virol. 61:3096-3101; Fisher et al. (1996), J. Virol., 70:520-532; Samulski et al. (1989), J. Virol. 63:3822-3826; U.S. Pat. No. 5,252,479; U.S. Pat. No. 5,139,941; International Patent Application No. WO 94/13788; and International Patent Application No. WO 93/24641, the entire disclosures of which are incorporated herein by reference. In one embodiment, the miR gene products are expressed from a single recombinant AAV vector comprising the CMV intermediate early promoter.
In a certain embodiment, a recombinant AAV viral vector of the invention comprises a nucleic acid sequence encoding a miR precursor RNA in operable connection with a polyT termination sequence under the control of a human U6 RNA promoter. As used herein, “in operable connection with a polyT termination sequence” means that the nucleic acid sequences encoding the sense or antisense strands are immediately adjacent to the polyT termination signal in the 5′ direction. During transcription of the miR sequences from the vector, the polyT termination signals act to terminate transcription.
In other embodiments of the treatment methods of the invention, an effective amount of at least one compound which inhibits miR expression can also be administered to the subject. As used herein, “inhibiting miR expression” means that the production of the active, mature form of miR gene product after treatment is less than the amount produced prior to treatment. One skilled in the art can readily determine whether miR expression has been inhibited in a cancer cell, using for example the techniques for determining miR transcript level discussed above for the diagnostic method Inhibition can occur at the level of gene expression (such as, by inhibiting transcription of a miR gene encoding the miR gene product) or at the level of processing (such as, by inhibiting processing of a miR precursor into a mature, active miR).
As used herein, an “effective amount” of a compound that inhibits miR expression is an amount sufficient to inhibit proliferation of a cancer cell in a subject suffering from a cancer associated with a cancer-associated chromosomal feature. One skilled in the art can readily determine an effective amount of an miR expression-inhibiting compound to be administered to a given subject, by taking into account factors, such as the size and weight of the subject; the extent of disease penetration; the age, health and sex of the subject; the route of administration; and whether the administration is regional or systemic.
For example, an effective amount of the expression-inhibiting compound can be based on the approximate weight of a tumor mass to be treated. The approximate weight of a tumor mass can be determined by calculating the approximate volume of the mass, wherein one cubic centimeter of volume is roughly equivalent to one gram. An effective amount based on the weight of a tumor mass can be between about 10-500 micrograms/gram of tumor mass, at least about 10 micrograms/gram of tumor mass, at least about 60 micrograms/gram of tumor mass, and at least about 100 micrograms/gram of tumor mass.
An effective amount of a compound that inhibits miR expression can also be based on the approximate or estimated body weight of a subject to be treated. Such effective amounts are administered parenterally or enterally, among others, as described herein. For example, an effective amount of the expression-inhibiting compound administered to a subject can range from about 5-3000 micrograms/kg of body weight, from about 700-1000 micrograms/kg of body weight, or it can be greater than about 1000 micrograms/kg of body weight.
One skilled in the art can also readily determine an appropriate dosage regimen for administering a compound that inhibits miR expression to a given subject. For example, an expression-inhibiting compound can be administered to the subject once (for example, as a single injection or deposition). Alternatively, an expression-inhibiting compound can be administered once or twice daily to a subject for a period of from about three to about twenty-eight days, more preferably from about seven to about ten days. In a particular dosage regimen, an expression-inhibiting compound is administered once a day for seven days. Where a dosage regimen comprises multiple administrations, it is understood that the effective amount of the expression-inhibiting compound administered to the subject can comprise the total amount of compound administered over the entire dosage regimen.
Suitable compounds for inhibiting miR gene expression include double-stranded RNA (such as short- or small-interfering RNA or “siRNA”), antisense nucleic acids, and enzymatic RNA molecules, such as ribozymes. Each of these compounds can be targeted to a given miR gene product and destroy or induce the destruction of the target miR gene product.
For example, expression of a given miR gene can be inhibited by inducing RNA interference of the miR gene with an isolated double-stranded RNA (“dsRNA”) molecule which has at least 90%, for example at least 95%, at least 98%, at least 99% or 100%, sequence homology with at least a portion of the miR gene product. In a particular embodiment, the dsRNA molecule is a “short or small interfering RNA” or “siRNA.”
siRNA useful in the present methods comprise short double-stranded RNA from about 17 nucleotides to about 29 nucleotides in length, preferably from about 19 to about 25 nucleotides in length. The siRNA comprise a sense RNA strand and a complementary antisense RNA strand annealed together by standard Watson-Crick base-pairing interactions (hereinafter “base-paired”). The sense strand comprises a nucleic acid sequence which is substantially identical to a nucleic acid sequence contained within the target miR gene product.
As used herein, a nucleic acid sequence in a siRNA which is “substantially identical” to a target sequence contained within the target mRNA is a nucleic acid sequence that is identical to the target sequence, or that differs from the target sequence by one or two nucleotides. The sense and antisense strands of the siRNA can comprise two complementary, single-stranded RNA molecules, or can comprise a single molecule in which two complementary portions are base-paired and are covalently linked by a single-stranded “hairpin” area.
The siRNA can also be altered RNA that differs from naturally-occurring RNA by the addition, deletion, substitution and/or alteration of one or more nucleotides. Such alterations can include addition of non-nucleotide material, such as to the end(s) of the siRNA or to one or more internal nucleotides of the siRNA, or modifications that make the siRNA resistant to nuclease digestion, or the substitution of one or more nucleotides in the siRNA with deoxyribonucleotides.
One or both strands of the siRNA can also comprise a 3′ overhang. As used herein, a “3′ overhang” refers to at least one unpaired nucleotide extending from the 3′-end of a duplexed RNA strand. Thus, in certain embodiments, the siRNA comprises at least one 3′ overhang of from 1 to about 6 nucleotides (which includes ribonucleotides or deoxyribonucleotides) in length, from 1 to about 5 nucleotides in length, from 1 to about 4 nucleotides in length, or from about 2 to about 4 nucleotides in length. In a particular embodiment, the 3′ overhang is present on both strands of the siRNA, and is 2 nucleotides in length. For example, each strand of the siRNA can comprise 3′ overhangs of dithymidylic acid (“TT”) or diuridylic acid (“uu”).
The siRNA can be produced chemically or biologically, or can be expressed from a recombinant plasmid or viral vector, as described above for the isolated miR gene products. Exemplary methods for producing and testing dsRNA or siRNA molecules are described in U.S. Published Patent Application No. 2002/0173478 to Gewirtz and in U.S. Pat. No. 7,148,342 to Reich et al., the entire disclosures of which are incorporated herein by reference.
Expression of a given miR gene can also be inhibited by an antisense nucleic acid. As used herein, an “antisense nucleic acid” refers to a nucleic acid molecule that binds to target RNA by means of RNA-RNA or RNA-DNA or RNA-peptide nucleic acid interactions, which alters the activity of the target RNA. Antisense nucleic acids suitable for use in the present methods are single-stranded nucleic acids (for example, RNA, DNA, RNA-DNA chimeras, PNA) that generally comprise a nucleic acid sequence complementary to a contiguous nucleic acid sequence in an miR gene product. The antisense nucleic acid can comprise a nucleic acid sequence that is 50-100% complementary, 75-100% complementary, or 95-100% complementary to a contiguous nucleic acid sequence in an miR gene product. Nucleic acid sequences for the miR gene products are provided in Table 1. Without wishing to be bound by any theory, it is believed that the antisense nucleic acids activate RNase H or another cellular nuclease that digests the miR gene product/antisense nucleic acid duplex.
Antisense nucleic acids can also contain modifications to the nucleic acid backbone or to the sugar and base moieties (or their equivalent) to enhance target specificity, nuclease resistance, delivery or other properties related to efficacy of the molecule. Such modifications include cholesterol moieties, duplex intercalators, such as acridine, or one or more nuclease-resistant groups.
Antisense nucleic acids can be produced chemically or biologically, or can be expressed from a recombinant plasmid or viral vector, as described above for the isolated miR gene products. Exemplary methods for producing and testing are within the skill in the art; see, for example, Stein and Cheng (1993), Science 261:1004 and U.S. Pat. No. 5,849,902 to Woolf et al., the entire disclosures of which are incorporated herein by reference.
Expression of a given miR gene can also be inhibited by an enzymatic nucleic acid. As used herein, an “enzymatic nucleic acid” refers to a nucleic acid comprising a substrate binding region that has complementarity to a contiguous nucleic acid sequence of an miR gene product, and which is able to specifically cleave the miR gene product. The enzymatic nucleic acid substrate binding region can be, for example, 50-100% complementary, 75-100% complementary, or 95-100% complementary to a contiguous nucleic acid sequence in a miR gene product. The enzymatic nucleic acids can also comprise modifications at the base, sugar, and/or phosphate groups. An exemplary enzymatic nucleic acid for use in the present methods is a ribozyme.
The enzymatic nucleic acids can be produced chemically or biologically, or can be expressed from a recombinant plasmid or viral vector, as described above for the isolated miR gene products. Exemplary methods for producing and testing dsRNA or siRNA molecules are described in Werner and Uhlenbeck (1995), Nucl. Acids Res. 23:2092-96; Hammann et al. (1999), Antisense and Nucleic Acid Drug Dev. 9:25-31; and U.S. Pat. No. 4,987,071 to Cech et al, the entire disclosures of which are incorporated herein by reference.
Administration of at least one miR gene product, or at least one compound for inhibiting miR expression, will inhibit the proliferation of cancer cells in a subject who has a cancer associated with a cancer-associated chromosomal feature. As used herein, to “inhibit the proliferation of a cancer cell” means to kill the cell, or permanently or temporarily arrest or slow the growth of the cell. Inhibition of cancer cell proliferation can be inferred if the number of such cells in the subject remains constant or decreases after administration of the miR gene products or miR gene expression-inhibiting compounds. An inhibition of cancer cell proliferation can also be inferred if the absolute number of such cells increases, but the rate of tumor growth decreases.
The number of cancer cells in a subject's body can be determined by direct measurement, or by estimation from the size of primary or metastatic tumor masses. For example, the number of cancer cells in a subject can be measured by immunohistological methods, flow cytometry, or other techniques designed to detect characteristic surface markers of cancer cells.
The size of a tumor mass can be ascertained by direct visual observation, or by diagnostic imaging methods, such as X-ray, magnetic resonance imaging, ultrasound, and scintigraphy. Diagnostic imaging methods used to ascertain size of the tumor mass can be employed with or without contrast agents, as is known in the art. The size of a tumor mass can also be ascertained by physical means, such as palpation of the tissue mass or measurement of the tissue mass with a measuring instrument, such as a caliper.
The miR gene products or miR gene expression-inhibiting compounds can be administered to a subject by any means suitable for delivering these compounds to cancer cells of the subject. For example, the miR gene products or miR expression inhibiting compounds can be administered by methods suitable to transfect cells of the subject with these compounds, or with nucleic acids comprising sequences encoding these compounds. In one embodiment, the cells are transfected with a plasmid or viral vector comprising sequences encoding at least one miR gene product or miR gene expression inhibiting compound.
Transfection methods for eukaryotic cells are well known in the art, and include, for example, direct injection of the nucleic acid into the nucleus or pronucleus of a cell; electroporation; liposome transfer or transfer mediated by lipophilic materials; receptor-mediated nucleic acid delivery, bioballistic or particle acceleration; calcium phosphate precipitation, and transfection mediated by viral vectors.
For example, cells can be transfected with a liposomal transfer compound, such as, DOTAP (N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethyl-ammonium methylsulfate, Boehringer-Mannheim) or an equivalent, such as LIPOFECTIN. The amount of nucleic acid used is not critical to the practice of the invention; acceptable results may be achieved with 0.1-100 micrograms of nucleic acid/10⁵cells. For example, a ratio of about 0.5 micrograms of plasmid vector in 3 micrograms of DOTAP per 10⁵cells can be used.
A miR gene product or miR gene expression inhibiting compound can also be administered to a subject by any suitable enteral or parenteral administration route. Suitable enteral administration routes for the present methods include, for example, oral, rectal, or intranasal delivery. Suitable parenteral administration routes include, for example, intravascular administration (for example, intravenous bolus injection, intravenous infusion, intra-arterial bolus injection, intra-arterial infusion and catheter instillation into the vasculature); peri- and intra-tissue injection (for example, peri-tumoral and intra-tumoral injection, intra-retinal injection, or subretinal injection); subcutaneous injection or deposition, including subcutaneous infusion (such as by osmotic pumps); direct application to the tissue of interest, for example by a catheter or other placement device (for example, a retinal pellet or a suppository or an implant comprising a porous, non-porous, or gelatinous material); and inhalation. Particularly suitable administration routes are injection, infusion and direct injection into the tumor.
In the present methods, a miR gene product or miR gene product expression inhibiting compound can be administered to the subject either as naked RNA, in combination with a delivery reagent, or as a nucleic acid (for example, a recombinant plasmid or viral vector) comprising sequences that express the miR gene product or expression inhibiting compound. Suitable delivery reagents include, for example, the Mirus Transit TKO lipophilic reagent; lipofectin; lipofectamine; cellfectin; polycations (for example, polylysine), and liposomes.
Recombinant plasmids and viral vectors comprising sequences that express the miR gene products or miR gene expression inhibiting compounds, and techniques for delivering such plasmids and vectors to cancer cells, are discussed herein.
In a particular embodiment, liposomes are used to deliver a miR gene product or miR gene expression-inhibiting compound (or nucleic acids comprising sequences encoding them) to a subject. Liposomes can also increase the blood half-life of the gene products or nucleic acids. Suitable liposomes for use in the invention can be formed from standard vesicle-forming lipids, which generally include neutral or negatively charged phospholipids and a sterol, such as cholesterol. The selection of lipids is generally guided by consideration of factors, such as the desired liposome size and half-life of the liposomes in the blood stream. A variety of methods are known for preparing liposomes, for example, as described in Szoka et al. (1980), Ann. Rev. Biophys. Bioeng. 9:467; and U.S. Pat. Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369, the entire disclosures of which are incorporated herein by reference.
The liposomes for use in the present methods can comprise a ligand molecule that targets the liposome to cancer cells. Ligands which bind to receptors prevalent in cancer cells, such as monoclonal antibodies that bind to tumor cell antigens, are preferred.
The liposomes for use in the present methods can also be modified so as to avoid clearance by the mononuclear macrophage system (“MMS”) and reticuloendothelial system (“RES”). Such modified liposomes have opsonization-inhibition moieties on the surface or incorporated into the liposome structure. In a particularly preferred embodiment, a liposome of the invention can comprise both opsonization-inhibition moieties and a ligand.
Opsonization-inhibiting moieties for use in preparing the liposomes of the invention are typically large hydrophilic polymers that are bound to the liposome membrane. As used herein, an opsonization inhibiting moiety is “bound” to a liposome membrane when it is chemically or physically attached to the membrane, for example, by the intercalation of a lipid-soluble anchor into the membrane itself, or by binding directly to active groups of membrane lipids. These opsonization-inhibiting hydrophilic polymers form a protective surface layer that significantly decreases the uptake of the liposomes by the MMS and RES; for example, as described in U.S. Pat. No. 4,920,016, the entire disclosure of which is incorporated herein by reference.
Opsonization inhibiting moieties suitable for modifying liposomes are preferably water-soluble polymers with a number-average molecular weight from about 500 to about 40,000 daltons, and more preferably from about 2,000 to about 20,000 daltons. Such polymers include polyethylene glycol (PEG) or polypropylene glycol (PPG) derivatives; for example, methoxy PEG or PPG, and PEG or PPG stearate; synthetic polymers, such as polyacrylamide or poly N-vinyl pyrrolidone; linear, branched, or dendrimeric polyamidoamines; polyacrylic acids; polyalcohols, for example, polyvinylalcohol and polyxylitol to which carboxylic or amino groups are chemically linked, as well as gangliosides, such as ganglioside GM1. Copolymers of PEG, methoxy PEG, or methoxy PPG, or derivatives thereof, are also suitable. In addition, the opsonization inhibiting polymer can be a block copolymer of PEG and either a polyamino acid, polysaccharide, polyamidoamine, polyethyleneamine, or polynucleotide. The opsonization inhibiting polymers can also be natural polysaccharides containing amino acids or carboxylic acids, for example, galacturonic acid, glucuronic acid, mannuronic acid, hyaluronic acid, pectic acid, neuraminic acid, alginic acid, carrageenan; aminated polysaccharides or oligosaccharides (linear or branched); or carboxylated polysaccharides or oligosaccharides, for example, reacted with derivatives of carbonic acids with resultant linking of carboxylic groups. Preferably, the opsonization-inhibiting moiety is a PEG, PPG, or derivatives thereof. Liposomes modified with PEG or PEG-derivatives are sometimes called “PEGylated liposomes.”
The opsonization inhibiting moiety can be bound to the liposome membrane by any one of numerous well-known techniques. For example, an N-hydroxysuccinimide ester of PEG can be bound to a phosphatidyl-ethanolamine lipid-soluble anchor, and then bound to a membrane. Similarly, a dextran polymer can be derivatized with a stearylamine lipid-soluble anchor via reductive amination using Na(CN)BH₃and a solvent mixture, such as tetrahydrofuran and water in a 30:12 ratio at 60° C.
Liposomes modified with opsonization-inhibition moieties remain in the circulation much longer than unmodified liposomes. For this reason, such liposomes are sometimes called “stealth” liposomes. Stealth liposomes are known to accumulate in tissues fed by porous or “leaky” microvasculature. Thus, tissue characterized by such microvasculature defects, for example solid tumors, will efficiently accumulate these liposomes; see Gabizon, et al. (1988), Proc. Natl. Acad. Sci., U.S.A., 18:6949-53. In addition, the reduced uptake by the RES lowers the toxicity of stealth liposomes by preventing significant accumulation of the liposomes in the liver and spleen. Thus, liposomes that are modified with opsonization-inhibition moieties are particularly suited to deliver the miR gene products or miR gene expression inhibition compounds (or nucleic acids comprising sequences encoding them) to tumor cells.
The miR gene products or miR gene expression inhibition compounds can be formulated as pharmaceutical compositions, sometimes called “medicaments,” prior to administering them to a subject, according to techniques known in the art. Accordingly, the invention encompasses pharmaceutical compositions for treating breast cancer. In one embodiment, the pharmaceutical compositions comprise at least one isolated miR gene product and a pharmaceutically-acceptable carrier. In a particular embodiment, the at least one miR gene product corresponds to a miR gene product that has a decreased level of expression in breast cancer cells relative to suitable control cells. In certain embodiments the isolated miR gene product is selected from the group consisting of miR-145, miR-10b, miR-123 (miR-126), miR-140-as, miR-125a, miR-125b-1, miR-125b-2, miR-194, miR-204, let-7a-2, let-7a-3, let-7d (let-7d-v1), let-7f-2, miR-101-1, miR-143 and combinations thereof.
In other embodiments, the pharmaceutical compositions of the invention comprise at least one miR expression inhibition compound. In a particular embodiment, the at least one miR gene expression inhibition compound is specific for a miR gene whose expression is greater in breast cancer cells than control cells. In certain embodiments, the miR gene expression inhibition compound is specific for one or more miR gene products selected from the group consisting of miR-21, miR-155, miR-009-1 (miR131-1), miR-34 (miR-170), miR-102 (miR-29b), miR-213, let-7i (let-7d-v2), miR-122a, miR-128b, miR-136, miR-149, miR-191, miR-196-1, miR-196-2, miR-202, miR-203, miR-206, miR-210, miR-213 and combinations thereof.
Pharmaceutical compositions of the present invention are characterized as being at least sterile and pyrogen-free. As used herein, “pharmaceutical formulations” include formulations for human and veterinary use. Methods for preparing pharmaceutical compositions of the invention are within the skill in the art, for example as described in Remington's Pharmaceutical Science, 17th ed., Mack Publishing Company, Easton, Pa. (1985), the entire disclosure of which is incorporated herein by reference.
The present pharmaceutical formulations comprise at least one miR gene product or miR gene expression inhibition compound (or at least one nucleic acid comprising sequences encoding them) (for example, 0.1 to 90% by weight), or a physiologically acceptable salt thereof, mixed with a pharmaceutically-acceptable carrier. The pharmaceutical formulations of the invention can also comprise at least one miR gene product or miR gene expression inhibition compound (or at least one nucleic acid comprising sequences encoding them) which are encapsulated by liposomes and a pharmaceutically-acceptable carrier. In one embodiment, the pharmaceutical compositions comprise a miR gene or gene product that is not miR-15, miR-16, miR-143 and/or miR-145.
Especially suitable pharmaceutically-acceptable carriers are water, buffered water, normal saline, 0.4% saline, 0.3% glycine, hyaluronic acid and the like.
In a particular embodiment, the pharmaceutical compositions of the invention comprise at least one miR gene product or miR gene expression inhibition compound (or at least one nucleic acid comprising sequences encoding them) which is resistant to degradation by nucleases. One skilled in the art can readily synthesize nucleic acids which are nuclease resistant, for example by incorporating one or more ribonucleotides that are modified at the 2′-position into the miR gene products. Suitable 2′-modified ribonucleotides include those modified at the 2′-position with fluoro, amino, alkyl, alkoxy, and O-allyl.
Pharmaceutical compositions of the invention can also comprise conventional pharmaceutical excipients and/or additives. Suitable pharmaceutical excipients include stabilizers, antioxidants, osmolality adjusting agents, buffers, and pH adjusting agents. Suitable additives include, for example, physiologically biocompatible buffers (for example, tromethamine hydrochloride), additions of chelants (such as, for example, DTPA or DTPA-bisamide) or calcium chelate complexes (such as, for example, calcium DTPA, CaNaDTPA-bisamide), or, optionally, additions of calcium or sodium salts (for example, calcium chloride, calcium ascorbate, calcium gluconate or calcium lactate). Pharmaceutical compositions of the invention can be packaged for use in liquid form, or can be lyophilized.
For solid pharmaceutical compositions of the invention, conventional nontoxic solid pharmaceutically-acceptable carriers can be used; for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, glucose, sucrose, magnesium carbonate, and the like.
For example, a solid pharmaceutical composition for oral administration can comprise any of the carriers and excipients listed above and 10-95%, preferably 25%-75%, of the at least one miR gene product or miR gene expression inhibition compound (or at least one nucleic acid comprising sequences encoding them). A pharmaceutical composition for aerosol (inhalational) administration can comprise 0.01-20% by weight, preferably 1%-10% by weight, of the at least one miR gene product or miR gene expression inhibition compound (or at least one nucleic acid comprising sequences encoding them) encapsulated in a liposome as described above, and a propellant. A carrier can also be included as desired; for example, lecithin for intranasal delivery.
The invention also encompasses methods of identifying an anti-breast cancer agent, comprising providing a test agent to a cell and measuring the level of at least one miR gene product in the cell. In one embodiment, the method comprises providing a test agent to a cell and measuring the level of at least one miR gene product associated with decreased expression levels in breast cancer cells. An increase in the level of the miR gene product in the cell, relative to a suitable control cell, is indicative of the test agent being an anti-breast cancer agent. In a particular embodiment, at least one miR gene product associated with decreased expression levels in breast cancer cells is selected from the group consisting of miR-145, miR-10b, miR-123 (miR-126), miR-140-as, miR-125a, miR-125b-1, miR-125b-2, miR-194, miR-204, let-7a-2, let-7a-3, let-7d (let-7d-v1), let-7f-2, miR-101-1, miR-143 and combinations thereof.
In other embodiments the method comprises providing a test agent to a cell and measuring the level of at least one miR gene product associated with increased expression levels in breast cancer cells. A decrease in the level of the miR gene product in the cell, relative to a suitable control cell, is indicative of the test agent being an anti-breast cancer agent. In a particular embodiment, at least one miR gene product associated with increased expression levels in breast cancer cells is selected from the group consisting of miR-21, miR-155, miR-009-1 (miR131-1), miR-34 (miR-170), miR-102 (miR-29b), miR-213, let-7i (let-7d-v2), miR-122a, miR-128b, miR-136, miR-149, miR-191, miR-196-1, miR-196-2, miR-202, miR-203, miR-206, miR-210, miR-213 and combinations thereof.
Suitable agents include, but are not limited to drugs (for example, small molecules, peptides), and biological macromolecules (for example, proteins, nucleic acids). The agent can be produced recombinantly, synthetically, or it may be isolated (i.e., purified) from a natural source. Various methods for providing such agents to a cell (for example, transfection) are well known in the art, and several of such methods are described hereinabove. Methods for detecting the expression of at least one miR gene product (for example, Northern blotting, in situ hybridization, RT-PCR, expression profiling) are also well known in the art. Several of these methods are also described hereinabove.
The invention will now be illustrated by the following non-limiting examples.

Example 1

Identification of a microRNA Expression Signature that Discriminates Breast Cancer Tissues from Normal Tissues

Materials and Methods
Breast Cancer Samples and Cell Lines.
RNAs from primary tumors were obtained from 76 samples collected at the University of Ferrara (Italy), Istituto Nazionale dei Tumori, Milano (Italy) and Thomas Jefferson University (Philadelphia, Pa.). Clinico-pathological information was available for 58 tumor samples. RNA from normal samples consisted of 6 pools of RNA from 5 normal breast tissues each, as well as RNA from 4 additional single breast tissues. Breast cancer RNAs were also obtained from the following cell lines: Hs578-T, MCF7, T47D, BT20, SK-BR-3, HBL100, HCC2218, MDA-MB-175, MDA-MB-231, MDA-MB-361, MDA-MB-435, MDA-MB-436, MDA-MB-453 and MDAMB-468.
miRNA Microarray.
Total RNA isolation was performed with Trizol Reagent (Invitrogen) according to the manufacturer's instructions. RNA labeling and hybridization on microRNA microarray chips was performed as previously described (Liu, C.-G., et al., Proc. Natl. Acad. Sci. U.S.A. 101:9740-9744 (2004)). Briefly, 5 μg of RNA from each sample was labeled with biotin during reverse transcription using random hexamers. Hybridization was carried out on a miRNA microarray chip (KCl version 1.0), which contains 368 probes, including 245 human and mouse miRNA genes, in triplicate. Hybridization signals were detected by binding of biotin to a Streptavidin-Alexa647 conjugate using a Perkin-Elmer ScanArray XL5K. Scanner images were quantified by the Quantarray software (Perkin Elmer).
Statistical and bioinformatic analysis of microarray data. Raw data were normalized and analyzed using the GeneSpring® software, version 7.2 (SiliconGenetics, Redwood City, Calif.). Expression data were median centered. Statistical comparisons were performed by ANOVA (Analysis of Variance), using the Benjamini and Hochberg correction for reduction of false positives. Prognostic miRNAs for tumor or normal class prediction were determined using both the PAM software (Prediction Analysis of Microarrays) (Tibshirani, R., et al. Proc. Natl. Acad. Sci. U.S.A. 99:6567-6572 (2002)) and the Support Vector Machine (Furey, T. S., et al. Bioinformatics 16: 906-914 (2000)) software. Both algorithms were used for Cross-validation and Test-set prediction. All data were submitted using MIAMExpress to the Array Express database.
Northern Blotting.
Northern blot analysis was performed as previously described (Calin, G. A., et al., Proc. Natl. Acad. Sci. U.S.A. 99:15524-29 (2002)). RNA samples (10 μg each) were electrophoresed on 15% acrylamide, 7 M urea Criterion pre-casted gels (Bio-Rad) and transferred onto Hybond-N+ membrane (Amersham Pharmacia Biotech). The hybridization was performed at 37° C. in 7% sodium dodecyl sulfate (SDS)/0.2M Na₂PO₄(pH 7.0) for 16 hours. Membranes were washed twice at 42° C. with 2× standard saline phosphate (0.18 M NaCl/10 mM phosphate, pH 7.4), supplemented with 1 mM EDTA (SSPE) and 0.1% SDS, and twice with 0.5×SSPE/0.1% SDS. Oligonucleotide probes were complementary to the sequence of the corresponding mature microRNA (see Sanger miR Registry): miR-21 5′-TCA ACA TCA GTC TGA TAA GCT A-3′ (SEQ ID NO:287); miR-125b1: 5′-TCA CAA GTT AGG GTC TCA GGG A-3′ (SEQ ID NO:288); miR-145: 5′-AAG GGA TTC CTG GGA AAA CTG GAC-3′ (SEQ ID NO:289). An oligonucleotide that was complementary to the U6 RNA (5′-GCA GGG GCC ATG CTA ATC TTC TCT GTA TCG-3′ (SEQ ID NO:290)) was used for normalizing expression levels. 200 μg of each probe was end labeled with 100 mCi [gamma-³²P]-ATP using a polynucleotide kinase (Roche). Northern Blots were stripped in a boiling 0.1% SDS solution for 10 minutes before re-hybridization.
Results
A microRNA microarray (Liu, C.-G., et al., Proc. Natl. Acad. Sci. U.S.A. 101:9740-9744 (2004)) was used to generate microRNA expression profiles for 10 normal and 76 neoplastic breast tissues. Each tumor sample was derived from a single specimen, while 6 of the 10 normal samples consisted of pools of RNA made from five different normal breast tissues. Hence, 34 normal breast samples were actually examined in the study.
To identify miRNAs that were differentially-expressed between normal and tumor samples, and, therefore, can be used to distinguish normal from cancerous breast tissues, analyses of variance and class prediction statistical tools were utilized. Results of the ANOVA analysis on normalized data generated a profile of differentially-expressed miRNAs (p<0.05) between normal and cancerous breast tissues (Table 2). Cluster analysis, based on differentially-expressed miRNA, generated a tree having a clear distinction between normal and cancer tissues (FIG. 1A).
To accurately identify a set of predictive miRNAs capable of differentiating normal from breast cancer tissues, we used Support Vector Machine (GeneSpring software) and PAM (Prediction Analysis of Micro arrays). Results from the two class prediction analyses largely overlapped (Table 3 and FIG. 1B). Among the miRNAs listed in Table 3, 11 of 15 have an ANOVA p-value of less than 0.05. To confirm the results obtained by microarray analysis, we performed Northern blot analysis to assess expression levels for a subset of microRNAs, namely, mir-125b, mir-145 and mir-21, that were differentially-expressed in normal and cancerous breast tissues. Northern blot analysis confirmed results obtained by microarray analysis. In many cases, expression differences appeared stronger than those anticipated by the microarray studies (FIG. 1C).

TABLE 2

miRNAs differentially-expressed between breast carcinoma and normal breast tissue.

Breast Cancer

Normal Breast

Median

Range

Median

Range

	P-value	Normalized	Min		Max	Normalized	Min		Max

let-7a-2	1.94E−02	1.67	0.96	-	6.21	2.30	1.34	-	5.00
let-7a-3	4.19E−02	1.26	0.81	-	3.79	1.58	1.02	-	2.91
let-7d (=7d-v1)	4.61E−03	0.90	0.59	-	1.54	1.01	0.83	-	1.25
let-7f-2	6.57E−03	0.84	0.51	-	1.58	0.92	0.76	-	1.03
let-7i (=let-7d-v2)	3.38E−02	2.05	1.02	-	7.49	1.53	1.01	-	3.47
mir-009-1 (mir-131-1)	9.12E−03	1.36	0.69	-	4.16	1.01	0.61	-	2.44
mir-010b	4.49E−02	1.11	0.69	-	4.79	1.70	0.96	-	6.32
mir-021	4.67E−03	1.67	0.66	-	26.43	1.08	0.80	-	2.31
mir-034 (mir-170)	1.06E−02	1.67	0.70	-	6.40	1.09	0.65	-	3.17
mir-101-1	4.15E−03	0.83	0.52	-	1.26	0.90	0.77	-	1.05
mir-122a	3.43E−03	2.21	0.93	-	8.08	1.48	1.06	-	3.67
mir-125a	3.28E−03	1.20	0.69	-	2.36	1.73	1.21	-	3.34
mir-125b-1	2.65E−02	1.30	0.55	-	8.68	2.87	1.45	-	18.38
mir-125b-2	2.33E−02	1.26	0.69	-	8.28	2.63	1.40	-	18.78
mir-126b	1.60E−02	1.12	0.68	-	7.34	1.02	0.89	-	1.27
mir-136	2.42E−03	1.32	0.74	-	10.26	1.05	0.76	-	1.47
mir-143	7.11E−03	0.87	0.68	-	1.33	0.98	0.81	-	1.17
mir-145	4.02E−03	1.52	0.92	-	8.46	3.61	1.85	-	14.45
mir-149	2.75E−02	1.11	0.53	-	1.73	1.03	0.83	-	1.22
mir-155(BIC)	1.24E−03	1.75	0.95	-	11.45	1.37	1.11	-	1.88
mir-191	4.26E−02	5.17	1.03	-	37.81	3.12	1.45	-	14.56
mir-196-1	1.07E−02	1.20	0.57	-	3.95	0.95	0.66	-	1.75
mir-196-2	1.16E−03	1.46	0.57	-	5.55	1.04	0.79	-	1.80
mir-202	1.25E−02	1.05	0.71	-	2.03	0.89	0.65	-	1.20
mir-203	4.06E−07	1.12	0.50	-	5.89	0.86	0.71	-	1.04
mir-204	2.15E−03	0.78	0.46	-	1.04	0.89	0.72	-	1.08
mir-206	1.42E−02	2.55	1.22	-	8.42	1.95	1.34	-	3.22
mir-210	6.40E−13	1.60	0.98	-	12.13	1.12	0.97	-	1.29
mir-213	1.08E−02	3.72	1.42	-	40.83	2.47	1.35	-	5.91

TABLE 3

Normal and tumor breast tissues class predictor microRNAs

	Median		SVM
	expression	ANOVA^a	prediction	PAM score^c	Chromos

miRNA name	Cancer	Normal	Probability	strength^b	Cancer	Normal	map

mir-009-1	1.36	1.01	0.0091	8.05	0.011	−0.102	1q22
mir-010b	1.11	1.70	0.0449	8.70	−0.032	0.299	2q31
mir-021	1.67	1.08	0.0047	10.20	0.025	−0.235	17q23.2
mir-034	1.67	1.09	0.0106	8.05	0.011	−0.106	1p36.22
mir-102 (mir-29b)	1.36	1.14	>0.10	8.92	0.000	−0.004	1q32.2-32.3
mir-123 (mir-126)	0.92	1.13	0.0940	9.13	−0.015	0.138	9q34
mir-125a	1.20	1.73	0.0033	8.99	−0.040	0.381	19q13.4
mir-125b-1	1.30	2.87	0.0265	14.78	−0.096	0.915	11q24.1
mir-125b-2	1.26	2.63	0.0233	17.62	−0.106	1.006	21q11.2
mir-140-as	0.93	1.10	0.0695	11.01	−0.005	0.050	16q22.1
mir-145	1.52	3.61	0.0040	12.93	−0.158	1.502	5q32-33
mir-155(BIC)	1.75	1.37	0.0012	10.92	0.003	−0.030	21q21
mir-194	0.96	1.09	>0.10	11.12	−0.025	0.234	1q41
mir-204	0.78	0.89	0.0022	8.10	−0.015	0.144	9q21.1
mir-213	3.72	2.47	0.0108	9.44	0.023	−0.220	1q31.3-q32.1

^aAnalysis of Variance (Welch t-test in Genespring software package) as calculated in Table 2.
^bSupport Vector Machine prediction analysis tool (from Genespring 7.2 software package). Prediction strengths are calculated as negative natural log of the probability to predict the observed number of samples, in one of the two classes, by chance. The higher is the score, the best is the prediction strength.
^cCentroid scores for the two classes of the Prediction Analysis of Microarrays (Tibshirani, R., et al. Proc. Natl. Acad. Sci. U.S.A. 99: 6567-6572 (2002)).

Of the 29 miRNAs whose expression is significantly (p<0.05) deregulated according to the microarray analysis, a set of 15 miRNAs were able to correctly predict the nature of the sample analyzed (i.e., normal vs. tumor) with 100% accuracy. Among the differentially-expressed miRNAs, miR-10b, miR-125b, miR145, miR-21 and miR-155 were the most consistently deregulated miRNAs in breast cancer samples. Three of these, namely, miR-10b, miR-125b and miR-145, were down-regulated, while the remaining two, miR-21 and miR-155, were up-regulated, suggesting that they might act as tumor suppressor genes or oncogenes, respectively.

Example 2

Determination of Putative Gene Targets of miRNAs that are Deregulated in Breast Cancer Tissues

At present, the lack of knowledge about bona fide miRNA gene targets hampers a full understanding of which biological functions are deregulated in cancers characterized by aberrant miRNA expression. To identify putative targets of the most significantly de-regulated miRNAs from our study: miR-10b, miR125b, miR-145, miR-21 and miR-155 (see Example 1), we utilized multiple computational approaches. In particular, the analysis was performed using three algorithms, miRanda, TargetScan and PicTar, which are used to predict human miRNA gene targets (Enright, A. J., et al. Genome Biol. 5:R1 (2003); Lewis, B. P. et al., Cell 115:787-798 (2003); Krek, A., et al., Nat. Genet. 37:495-500 (2005)). The results obtained using each of the three algorithms were cross-referenced with one another to validate putative targets, and only targets that were identified by at least 2 of the 3 algorithms were considered. Results of this analysis are presented in Table 4.
Several genes with potential oncogenic functions were identified as putative targets of miRNAs that are down-regulated in breast cancer samples. Notably, oncogenes were identified as targets of miR-10b (for example, FLT1, the v-crk homolog, the growth factor BDNF and the transducing factor SHC1), miR-125b (for example, YES, ETS1, TEL, AKT3, the growth factor receptor FGFR2 and members of the mitogen-activated signal transduction pathway VTS58635, MAP3K10, MAP3K11, MAPK14), and miR-145 (for example, MYCN, FOS, YES and FLI1, integration site of Friend leukemia virus, cell cycle promoters, such as cyclins D2 and L1, MAPK transduction proteins, such as MAP3K3 and MAP4K4). The proto-oncogene, YES, and the core-binding transcription factor, CBFB, were determined to be potential targets of both miR-125 and miR-145.
Consistent with these findings, multiple tumor suppressor genes were identified as targets of miR-21 and miR-155, miRNAs that are up-regulated in breast cancer cells. For miR-21, the TGFB gene was predicted as target by all three methods. For miR-155, potential targets included the tumor suppressor genes, SOCS1 and APC, and the kinase, WEE1, which blocks the activity of Cdc2 and prevents entry into mitosis. The hypoxia inducible factor, HIF1A, was also a predicted target of miR-155. Notably, the tripartite motif-containing protein TRIM2, the proto-oncogene, SKI, and the RAS homologs, RAB6A and RAB6C, were found as potential targets of both miR-21 and miR-155.

TABLE 4

Putative gene targets of differentially-expressed miRNA identified by at least two prediction methods

				Prediction
miRNA	Genbank	Gene Symbol	Gene Name	algorithm	Gene Ontology condensed

miR-10b	AL117516	38596	strand-exchange protein 1	P + T	exonuclease activity\|nucleus
miR-10b	NM_004915	ABCG1	ATP-binding cassette, sub-	P + T	ATP binding\|ATPase
			family G (WHITE), member 1		activity\|ATPase activity, coupled to
					transmembrane movement of
					substances\|L-tryptophan transporter
					activity\|cholesterol
					homeostasis\|cholesterol
					metabolism\|detection of hormone
					stimulus\|integral to plasma
					membrane\|lipid
					transport\|membrane\|membrane
					fraction\|permease activity\|protein
					dimerization activity\|purine
					nucleotide transporter
					activity\|response to organic
					substance
miR-10b	NM_001148	ANK2	ankyrin 2, neuronal	P + T	actin
					cytoskeleton\|membrane\|metabolism\|
					oxidoreductase activity\|protein
					binding\|signal transduction\|structural
					constituent of cytoskeleton
miR-10b	NM_020987	ANK3	ankyrin 3, node of Ranvier	P + T	Golgi apparatus\|cytoskeletal
			(ankyrin G)		anchoring\|cytoskeleton\|cytoskeleton\|
					endoplasmic reticulum\|protein
					binding\|protein targeting\|signal
					transduction\|structural constituent of
					cytoskeleton
miR-10b	NM_016376	ANKHZN	ANKHZN protein	P + T	endocytosis\|endosome
					membrane\|membrane\|protein
					binding\|zinc ion binding
miR-10b	NM_006380	APPBP2	amyloid beta precursor	P + T	binding\|cytoplasm\|intracellular
			protein (cytoplasmic tail)		protein
			binding protein 2		transport\|membrane\|microtubule
					associated complex\|microtubule
					motor activity\|nucleus
miR-10b	NM_006321	ARIH2	ariadne homolog 2	P + T	development\|nucleic acid
			(Drosophila)		binding\|nucleus\|protein
					ubiquitination\|ubiquitin ligase
					complex\|ubiquitin-protein ligase
					activity\|zinc ion binding
miR-10b	NM_001668	ARNT	aryl hydrocarbon receptor	P + T	aryl hydrocarbon receptor nuclear
			nuclear translocator		translocator
					activity\|nucleus\|nucleus\|protein-
					nucleus import,
					translocation\|receptor
					activity\|regulation of transcription,
					DNA-dependent\|signal transducer
					activity\|signal
					transduction\|transcription coactivator
					activity\|transcription factor
					activity\|transcription factor activity
miR-10b	AI829840	ASXL1	ESTs, Weakly similar to	P + T	nucleus\|regulation of transcription,
			SFRB_HUMAN Splicing		DNA-dependent\|transcription
			factor arginine/serine-rich
			11 (Arginine-rich 54 kDa
			nuclear protein) (P54)
			[H. sapiens]
miR-10b	NM_021813	BACH2	BTB and CNC homology 1,	P + T	DNA binding\|nucleus\|protein
			basic leucine zipper		binding\|regulation of transcription,
			transcription factor 2		DNA-dependent\|transcription
miR-10b	NM_013450	BAZ2B	bromodomain adjacent to	P + T	DNA binding\|nucleus\|regulation of
			zinc finger domain, 2B		transcription, DNA-
					dependent\|transcription
miR-10b	NM_001706	BCL6	B-cell CLL/lymphoma 6	P + T	inflammatory response\|mediator
			(zinc finger protein 51)		complex\|negative regulation of
					transcription from RNA polymerase
					II promoter\|nucleus\|positive
					regulation of cell
					proliferation\|protein
					binding\|regulation of transcription,
					DNA-
					dependent\|transcription\|transcription
					factor activity\|zinc ion binding
miR-10b	NM_001709	BDNF	brain-derived neurotrophic	P + T	growth factor activity\|growth factor
			factor		activity\|neurogenesis
miR-10b	NM_006624	BS69	adenovirus 5 E1A binding	P + T	DNA binding\|cell cycle\|cell
			protein		proliferation\|negative regulation of
					cell cycle\|negative regulation of
					transcription from RNA polymerase
					II promoter\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription
miR-10b	AF101784	BTRC	beta-transducin repeat	P + T	Wnt receptor signaling
			containing		pathway\|endoplasmic
					reticulum\|ligase activity\|signal
					transduction\|ubiquitin conjugating
					enzyme activity\|ubiquitin
					cycle\|ubiquitin-dependent protein
					catabolism
miR-10b	NM_005808	C3orf8	HYA22 protein	P + T	biological_process
					unknown\|molecular_function
					unknown\|nucleus
miR-10b	BF111268	CAMK2G	calcium/calmodulin-	P + T	ATP binding\|ATP binding\|calcium-
			dependent protein kinase		and calmodulin-dependent protein
			(CaM kinase) II gamma		kinase activity\|calcium-dependent
					protein serine/threonine phosphatase
					activity\|calmodulin
					binding\|cellular_component
					unknown\|insulin secretion\|kinase
					activity\|protein amino acid
					phosphorylation\|protein amino acid
					phosphorylation\|protein
					serine/threonine kinase
					activity\|protein-tyrosine kinase
					activity\|signal
					transduction\|transferase activity
miR-10b	NM_020184	CNNM4	cyclin M4	P + T
miR-10b	NM_022730	COPS7B	COP9 constitutive	P + T	signalosome complex
			photomorphogenic homolog
			subunit 7B (Arabidopsis)
miR-10b	NM_016823	CRK	v-crk sarcoma virus CT10	P + T	SH3/SH2 adaptor activity\|actin
			oncogene homolog (avian)		cytoskeleton organization and
					biogenesis\|cell
					motility\|cytoplasm\|intracellular
					signaling cascade\|nucleus\|regulation
					of transcription from RNA
					polymerase II promoter
miR-10b	NM_020248	CTNNBIP1	catenin, beta interacting	P + T	Wnt receptor signaling pathway\|beta-
			protein 1		catenin binding\|cell
					proliferation\|development\|nucleus\|regulation
					of transcription, DNA-
					dependent\|signal transduction
miR-10b	NM_018959	DAZAP1	DAZ associated protein 1	P + T	RNA binding\|cell
					differentiation\|nucleotide
					binding\|nucleus\|spermatogenesis
miR-10b	AL136828	DKFZP434K0427	hypothetical protein	P + T	cation transport\|cation transporter
			DKFZp434K0427		activity
miR-10b	R20763	DKFZp547J036	ELAV (embryonic lethal,	P + T
			abnormal vision,
			Drosophila)-like 3 (Hu
			antigen C)
miR-10b	AF009204	DLGAP2	discs, large (Drosophila)	P + T	cell-cell signaling\|membrane\|nerve-
			homolog-associated protein 2		nerve synaptic
					transmission\|neurofilament\|protein
					binding
miR-10b	NM_001949	E2F3	E2F transcription factor 3	P + T	nucleus\|protein binding\|regulation of
					cell cycle\|regulation of transcription,
					DNA-
					dependent\|transcription\|transcription
					factor activity\|transcription factor
					complex\|transcription initiation from
					RNA polymerase II promoter
miR-10b	NM_022659	EBF2	early B-cell factor 2	P + T	DNA
					binding\|development\|nucleus\|regulation
					of transcription, DNA-
					dependent\|transcription
miR-10b	NM_004432	ELAVL2	ELAV (embryonic lethal,	P + T	RNA binding\|mRNA 3′-UTR
			abnormal vision,		binding\|nucleotide binding\|regulation
			Drosophila)-like 2 (Hu		of transcription, DNA-dependent
			antigen B)
miR-10b	NM_001420	ELAVL3	ELAV (embryonic lethal,	P + T	RNA binding\|cell
			abnormal vision,		differentiation\|mRNA 3′-UTR
			Drosophila)-like 3 (Hu		binding\|neurogenesis\|nucleotide
			antigen C)		binding
miR-10b	NM_004438	EPHA4	EphA4	P + T	ATP binding\|ephrin receptor
					activity\|integral to plasma
					membrane\|membrane\|protein amino
					acid phosphorylation\|receptor
					activity\|signal
					transduction\|transferase
					activity\|transmembrane receptor
					protein tyrosine kinase signaling
					pathway
miR-10b	AL035703	EPHA8; EEK;	EphA8	P + T
		HEK3; Hek3;
		KIAA1459
miR-10b	NM_004468	FHL3	four and a half LIM	P + T	muscle development\|zinc ion binding
			domains 3
miR-10b	NM_024679	FLJ11939	hypothetical protein	P + T
			FLJ11939
miR-10b	AI742838	FLJ32122	hypothetical protein	P + T	GTP binding\|GTPase
			FLJ32122		binding\|guanyl-nucleotide exchange
					factor activity
miR-10b	AL040935	FLJ33957	hypothetical protein	P + T	protein binding
			FLJ33957
miR-10b	AA058828	FLT1	ESTs	P + T	ATP binding\|angiogenesis\|cell
					differentiation\|extracellular
					space\|integral to plasma
					membrane\|membrane\|positive
					regulation of cell
					proliferation\|pregnancy\|protein
					amino acid phosphorylation\|receptor
					activity\|transferase
					activity\|transmembrane receptor
					protein tyrosine kinase signaling
					pathway\|vascular endothelial growth
					factor receptor activity
miR-10b	NM_004860	FXR2	fragile X mental retardation,	P + T	RNA binding\|cytoplasm\|cytosolic
			autosomal homolog 2		large ribosomal subunit (sensu
					Eukaryota)\|nucleus
miR-10b	NM_020474	GALNT1	UDP-N-acetyl-alpha-D-	P + T	Golgi apparatus\|O-linked
			galactosamine:polypeptide		glycosylation\|integral to
			N-acetylgalactosaminyl-		membrane\|manganese ion
			transferase 1		binding\|polypeptide N-
			(GalNAc-T1)		acetylgalactosaminyltransferase
					activity\|sugar binding\|transferase
					activity, transferring glycosyl groups
miR-10b	D87811	GATA6	GATA binding protein 6	P + T	muscle development\|nucleus\|positive
					regulation of transcription\|regulation
					of transcription, DNA-
					dependent\|transcription\|transcription
					factor activity\|transcriptional
					activator activity\|zinc ion binding
miR-10b	NM_000840	GRM3	glutamate receptor,	P + T	G-protein coupled receptor protein
			metabotropic 3		signaling pathway\|integral to plasma
					membrane\|membrane\|metabotropic
					glutamate, GABA-B-like receptor
					activity\|negative regulation of
					adenylate cyclase activity\|receptor
					activity\|signal transduction\|synaptic
					transmission
miR-10b	NM_005316	GTF2H1	general transcription factor	P + T	DNA repair\|[RNA-polymerase]-
			IIH, polypeptide 1, 62 kDa		subunit kinase activity\|general RNA
					polymerase II transcription factor
					activity\|nucleus\|regulation of cyclin
					dependent protein kinase
					activity\|regulation of transcription,
					DNA-
					dependent\|transcription\|transcription
					factor TFIIH complex\|transcription
					from RNA polymerase II promoter
miR-10b	AF232772	HAS3	hyaluronan synthase 3	P + T	carbohydrate metabolism\|hyaluronan
					synthase activity\|integral to plasma
					membrane\|transferase activity,
					transferring glycosyl groups
miR-10b	AL023584	HIVEP2	human immunodeficiency	P + T
			virus type I enhancer
			binding protein 2
miR-10b	S79910	HOXA1	homeo box A1	P + T	RNA polymerase II transcription
					factor
					activity\|development\|nucleus\|regulation
					of transcription, DNA-
					dependent\|transcription factor
					activity
miR-10b	NM_030661	HOXA3	homeo box A3	P + T	development\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription factor
					activity
miR-10b	AW299531	HOXD10	homeo box D10	P + T	RNA polymerase II transcription
					factor
					activity\|development\|nucleus\|regulation
					of transcription, DNA-
					dependent\|transcription factor
					activity
miR-10b	BF031714	HYA22	HYA22 protein	P + T
miR-10b	NM_001546	ID4	inhibitor of DNA binding 4,	P + T	nucleus\|regulation of transcription
			dominant negative helix-		from RNA polymerase II
			loop-helix protein		promoter\|transcription corepressor
					activity
miR-10b	NM_014333	IGSF4	immunoglobulin	P + T
			superfamily, member 4
miR-10b	NM_014271	IL1RAPL1	interleukin 1 receptor	P + T	integral to membrane\|learning and/or
			accessory protein-like 1		memory\|membrane\|signal
					transduction\|transmembrane receptor
					activity
miR-10b	D87450	KIAA0261	KIAA0261 protein	P + T
miR-10b	AL117518	KIAA0978	KIAA0978 protein	P + T	nucleus\|regulation of transcription,
					DNA-dependent\|transcription
miR-10b	AK025960	KIAA1255	KIAA1255 protein	P + T	endocytosis\|endosome
					membrane\|membrane\|protein
					binding\|zinc ion binding
miR-10b	AB037797	KIAA1376	KIAA1376 protein	P + T
miR-10b	NM_004795	KL	klotho	P + T	beta-glucosidase
					activity\|carbohydrate
					metabolism\|extracellular
					space\|glucosidase activity\|integral to
					membrane\|integral to plasma
					membrane\|membrane fraction\|signal
					transducer activity\|soluble fraction
miR-10b	NM_015995	KLF13	Kruppel-like factor 13	P + T	DNA binding\|RNA polymerase II
					transcription factor
					activity\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					from RNA polymerase II
					promoter\|zinc ion binding
miR-10b	NM_004235	KLF4	Kruppel-like factor 4 (gut)	P + T	mesodermal cell fate
					determination\|negative regulation of
					cell proliferation\|negative regulation
					of transcription, DNA-
					dependent\|negative regulation of
					transcription, DNA-
					dependent\|nucleic acid
					binding\|nucleus\|transcription\|transcription
					factor activity\|transcription
					factor activity\|transcriptional
					activator activity\|transcriptional
					activator activity\|transcriptional
					repressor activity\|transcriptional
					repressor activity\|zinc ion
					binding\|zinc ion binding
miR-10b	AW511293	LOC144455	hypothetical protein	P + T	regulation of cell cycle\|regulation of
			BC016658		transcription, DNA-
					dependent\|transcription factor
					activity\|transcription factor complex
miR-10b	NM_014921	LPHN1	lectomedin-2	P + T	G-protein coupled receptor
					activity\|integral to
					membrane\|latrotoxin receptor
					activity\|membrane\|neuropeptide
					signaling pathway\|receptor
					activity\|signal transduction\|sugar
					binding
miR-10b	NM_012325	MAPRE1	microtubule-associated	P + T	cell
			protein, RP/EB family,		proliferation\|cytokinesis\|microtubule
			member 1		binding\|mitosis\|protein C-terminus
					binding\|regulation of cell cycle
miR-10b	AA824369	MGC4643	hypothetical protein	P + T	Wnt receptor signaling
			MGC4643		pathway\|endoplasmic
					reticulum\|ligase activity\|signal
					transduction\|ubiquitin conjugating
					enzyme activity\|ubiquitin
					cycle\|ubiquitin-dependent protein
					catabolism
miR-10b	NM_021090	MTMR3	myotubularin related protein 3	P + T	cytoplasm\|hydrolase activity\|inositol
					or phosphatidylinositol phosphatase
					activity\|membrane\|membrane
					fraction\|phospholipid
					dephosphorylation\|protein amino
					acid dephosphorylation\|protein
					serine/threonine phosphatase
					activity\|protein tyrosine phosphatase
					activity\|protein
					tyrosine/serine/threonine
					phosphatase activity\|zinc ion binding
miR-10b	AI498126	NAC1	transcriptional repressor	P + T	protein binding
			NAC1
miR-10b	AF128458	NCOA6	nuclear receptor coactivator 6	P + T	DNA recombination\|DNA
					repair\|DNA replication\|brain
					development\|chromatin
					binding\|embryonic development
					(sensu Mammalia)\|estrogen receptor
					binding\|estrogen receptor signaling
					pathway\|glucocorticoid receptor
					signaling pathway\|heart
					development\|ligand-dependent
					nuclear receptor transcription
					coactivator activity\|myeloid blood
					cell
					differentiation\|nucleus\|nucleus\|positive
					regulation of transcription from
					RNA polymerase II promoter\|protein
					binding\|regulation of transcription,
					DNA-dependent\|response to
					hormone stimulus\|retinoid X receptor
					binding\|thyroid hormone receptor
					binding\|transcription\|transcription
					factor complex\|transcription
					initiation from RNA polymerase II
					promoter\|transcriptional activator
					activity
miR-10b	NM_006312	NCOR2	nuclear receptor corepressor 2	P + T	DNA binding\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription corepressor
					activity
miR-10b	NM_006599	NFAT5	nuclear factor of activated	P + T	RNA polymerase II transcription
			T-cells 5, tonicity-		factor
			responsive		activity\|excretion\|nucleus\|regulation
					of transcription, DNA-
					dependent\|signal
					transduction\|transcription factor
					activity\|transcription from RNA
					polymerase II promoter
miR-10b	NM_006981	NR4A3	nuclear receptor subfamily	M + P + T	binding\|nucleus\|nucleus\|regulation of
			4, group A, member 3		transcription, DNA-
					dependent\|steroid hormone receptor
					activity\|steroid hormone receptor
					activity\|thyroid hormone receptor
					activity\|transcription\|transcription
					factor activity
miR-10b	NM_003822	NR5A2	nuclear receptor subfamily	P + T	RNA polymerase II transcription
			5, group A, member 2		factor activity, enhancer
					binding\|morphogenesis\|nucleus\|nucleus\|
					regulation of transcription, DNA-
					dependent\|steroid hormone receptor
					activity\|transcription\|transcription
					factor activity\|transcription from
					RNA polymerase II promoter
miR-10b	AA295257	NRP2	neuropilin 2	P + T	angiogenesis\|axon guidance\|cell
					adhesion\|cell adhesion\|cell
					differentiation\|electron
					transport\|electron transporter
					activity\|integral to membrane\|integral
					to membrane\|membrane\|membrane
					fraction\|neurogenesis\|receptor
					activity\|semaphorin receptor
					activity\|vascular endothelial growth
					factor receptor activity\|vascular
					endothelial growth factor receptor
					activity
miR-10b	NM_000430	PAFAH1B1	platelet-activating factor	P + T	astral microtubule\|cell cortex\|cell
			acetylhydrolase, isoform Ib,		cycle\|cell differentiation\|cell
			alpha subunit 45 kDa		motility\|cytokinesis\|cytoskeleton\|dynein
					binding\|establishment of mitotic
					spindle orientation\|kinetochore\|lipid
					metabolism\|microtubule associated
					complex\|microtubule-based
					process\|mitosis\|neurogenesis\|nuclear
					membrane\|signal transduction
miR-10b	NM_013382	POMT2	putative protein O-	P + T	O-linked glycosylation\|dolichyl-
			mannosyltransferase		phosphate-mannose-protein
					mannosyltransferase
					activity\|endoplasmic
					reticulum\|integral to
					membrane\|magnesium ion
					binding\|membrane\|transferase
					activity, transferring glycosyl groups
miR-10b	BF337790	PURB	purine-rich element binding	P + T
			protein B
miR-10b	AI302106	RAP2A	RAP2A, member of RAS	P + T	GTP binding\|GTPase
			oncogene family		activity\|membrane\|signal
					transduction\|small GTPase mediated
					signal transduction
miR-10b	NM_002886	RAP2B	RAP2B, member of RAS	P + T	GTP binding\|protein transport\|small
			oncogene family		GTPase mediated signal transduction
miR-10b	NM_014781	RB1CC1	RB1-inducible coiled-coil 1	P + T	kinase activity
miR-10b	NM_012234	RYBP	RING1 and YY1 binding	P + T	development\|negative regulation of
			protein		transcription from RNA polymerase
					II promoter\|nucleus\|transcription
					corepressor activity
miR-10b	NM_005506	SCARB2	scavenger receptor class B,	P + T	cell adhesion\|integral to plasma
			member 2		membrane\|lysosomal
					membrane\|membrane
					fraction\|receptor activity
miR-10b	AF225986	SCN3A	sodium channel, voltage-	P + T	cation channel activity\|cation
			gated, type III, alpha		transport\|integral to
			polypeptide		membrane\|membrane\|sodium ion
					transport\|voltage-gated sodium
					channel activity\|voltage-gated
					sodium channel complex
miR-10b	NM_002997	SDC1	syndecan 1	P + T	cytoskeletal protein binding\|integral
					to plasma membrane\|membrane
miR-10b	NM_006924	SFRS1	splicing factor,	P + T	RNA binding\|mRNA splice site
			arginine/serine-rich 1		selection\|nuclear mRNA splicing, via
			(splicing factor 2, alternate		spliceosome\|nucleotide
			splicing factor)		binding\|nucleus
miR-10b	AI809967	SHC1	SHC (Src homology 2	P + T	activation of MAPK\|activation of
			domain containing)		MAPK\|intracellular signaling
			transforming protein 1		cascade\|phospholipid
					binding\|phospholipid binding\|plasma
					membrane\|plasma
					membrane\|positive regulation of cell
					proliferation\|positive regulation of
					cell proliferation\|positive regulation
					of mitosis\|positive regulation of
					mitosis\|regulation of cell
					growth\|regulation of epidermal
					growth factor receptor
					activity\|transmembrane receptor
					protein tyrosine kinase adaptor
					protein activity\|transmembrane
					receptor protein tyrosine kinase
					adaptor protein activity
miR-10b	NM_018976	SLC38A2	solute carrier family 38,	P + T	amino acid transport\|amino acid-
			member 2		polyamine transporter
					activity\|integral to
					membrane\|membrane\|oxygen
					transport\|oxygen transporter
					activity\|transport
miR-10b	NM_003794	SNX4	sorting nexin 4	P + T	endocytosis\|intracellular signaling
					cascade\|protein transport
miR-10b	NM_003103	SON	SON DNA binding protein	P + T	DNA binding\|DNA binding\|anti-
					apoptosis\|double-stranded RNA
					binding\|intracellular\|nucleic acid
					binding\|nucleus
miR-10b	Z48199		syndecan-1	P + T
miR-10b	NM_003222	TFAP2C	transcription factor AP-2	P + T	cell-cell signaling\|nucleus\|regulation
			gamma (activating enhancer		of transcription from RNA
			binding protein 2 gamma)		polymerase II
					promoter\|transcription\|transcription
					factor activity
miR-10b	NM_003275	TMOD1	tropomodulin	P + T	actin
					binding\|cytoskeleton\|cytoskeleton
					organization and
					biogenesis\|tropomyosin binding
miR-10b	NM_003367	USF2	upstream transcription factor	P + T	RNA polymerase II transcription
			2, c-fos interacting		factor activity\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					factor activity
miR-10b	N62196	ZNF367	zinc finger protein 367	P + T	nucleic acid binding\|nucleus\|zinc ion
					binding
miR-125b	AI948503	ABCC4	ATP-binding cassette, sub-	P + T	15-hydroxyprostaglandin
			family C (CFTR/MRP),		dehydrogenase (NAD+) activity\|ATP
			member 4		binding\|ATPase activity\|ATPase
					activity, coupled to transmembrane
					movement of substances\|chloride
					channel activity\|integral to
					membrane\|ion transport\|membrane
miR-125b	AL534702	ABHD3	abhydrolase domain	M + P + T
			containing 3
miR-125b	AL527773	ABR	active BCR-related gene	P + T	GTPase activator activity\|guanyl-
					nucleotide exchange factor
					activity\|small GTPase mediated
					signal transduction
miR-125b	NM_020039	ACCN2	amiloride-sensitive cation	P + T	amiloride-sensitive sodium channel
			channel 2, neuronal		activity\|integral to plasma
					membrane\|ion channel activity\|ion
					transport\|membrane\|response to
					pH\|signal transduction\|sodium ion
					transport
miR-125b	NM_003816	ADAM9	a disintegrin and	P + T	SH3 domain binding\|integral to
			metalloproteinase domain 9		plasma membrane\|integrin
			(meltrin gamma)		binding\|metalloendopeptidase
					activity\|protein binding\|protein
					kinase binding\|protein kinase
					cascade\|proteolysis and
					peptidolysis\|zinc ion binding
miR-125b	L05500	ADCY1	adenylate cyclase 1 (brain)	P + T	cAMP biosynthesis\|calcium- and
					calmodulin-responsive adenylate
					cyclase activity\|calmodulin
					binding\|integral to
					membrane\|intracellular signaling
					cascade\|magnesium ion binding
miR-125b	NM_017488	ADD2	adducin 2 (beta)	P + T	actin binding\|actin
					cytoskeleton\|calmodulin
					binding\|membrane
miR-125b	NM_003488	AKAP1	A kinase (PRKA) anchor	P + T	RNA binding\|integral to
			protein 1		membrane\|mitochondrion\|outer
					membrane
miR-125b	NM_005465	AKT3	v-akt murine thymoma viral	P + T	ATP binding\|protein amino acid
			oncogene homolog 3		phosphorylation\|protein
			(protein kinase B, gamma)		serine/threonine kinase
					activity\|signal
					transduction\|transferase activity
miR-125b	NM_001150	ANPEP	alanyl (membrane)	P + T	aminopeptidase
			aminopeptidase		activity\|angiogenesis\|cell
			(aminopeptidase N,		differentiation\|integral to plasma
			aminopeptidase M,		membrane\|membrane alanyl
			microsomal aminopeptidase,		aminopeptidase
			CD13, p150)		activity\|metallopeptidase
					activity\|proteolysis and
					peptidolysis\|receptor activity\|zinc ion
					binding
miR-125b	AF193759	APBA2BP	amyloid beta (A4) precursor	M + P + T	Golgi cis cisterna\|Golgi cis
			protein-binding, family A,		cisterna\|antibiotic
			member 2 binding protein		biosynthesis\|calcium ion
					binding\|cytoplasm\|cytoplasm\|endoplasmic
					reticulum
					membrane\|endoplasmic reticulum
					membrane\|nucleus\|oxidoreductase
					activity\|protein binding\|protein
					binding\|protein binding\|protein
					metabolism\|protein
					metabolism\|protein secretion\|protein
					secretion\|regulation of amyloid
					precursor protein biosynthesis
miR-125b	NM_000038	APC	adenomatosis polyposis coli	P + T	Wnt receptor signaling pathway\|beta-
					catenin binding\|cell
					adhesion\|microtubule
					binding\|negative regulation of cell
					cycle\|protein complex
					assembly\|signal transduction
miR-125b	NM_001655	ARCN1	archain 1	P + T	COPI vesicle coat\|Golgi
					apparatus\|clathrin vesicle coat\|intra-
					Golgi transport\|intracellular protein
					transport\|intracellular protein
					transport\|membrane\|retrograde
					transport, Golgi to ER\|transport
miR-125b	BC001719	ASB6	ankyrin repeat and SOCS	M + P	intracellular signaling cascade
			box-containing 6
miR-125b	AI478147	ATP10D	ATPase, Class V, type 10D	P + T	ATP binding\|ATPase activity\|cation
					transport\|hydrolase activity\|integral
					to membrane\|magnesium ion
					binding\|membrane\|phospholipid-
					translocating ATPase activity
miR-125b	NM_012069	ATP1B4	ATPase, (Na+)/K+	P + T	hydrogen ion transporter
			transporting, beta 4		activity\|integral to plasma
			polypeptide		membrane\|ion
					transport\|membrane\|potassium ion
					transport\|proton transport\|sodium ion
					transport\|sodium:potassium-
					exchanging ATPase activity
miR-125b	NM_005176	ATP5G2	ATP synthase, H+	M + P + T	ATP synthesis coupled proton
			transporting, mitochondrial		transport\|hydrogen-transporting ATP
			F0 complex, subunit c		synthase activity, rotational
			(subunit 9), isoform 2		mechanism\|hydrogen-transporting
					ATPase activity, rotational
					mechanism\|ion transport\|lipid
					binding\|membrane\|membrane
					fraction\|mitochondrion\|proton
					transport\|proton-transporting ATP
					synthase complex (sensu
					Eukaryota)\|proton-transporting two-
					sector ATPase complex\|transporter
					activity
miR-125b	NM_001702	BAI1	brain-specific angiogenesis	M + P + T	G-protein coupled receptor
			inhibitor
1		activity\|axonogenesis\|brain-specific
					angiogenesis inhibitor activity\|cell
					adhesion\|integral to plasma
					membrane\|intercellular
					junction\|negative regulation of cell
					proliferation\|neuropeptide signaling
					pathway\|peripheral nervous system
					development\|plasma
					membrane\|protein binding\|receptor
					activity\|signal transduction
miR-125b	NM_001188	BAK1	BCL2-antagonist/killer 1	M + T	apoptotic mitochondrial
					changes\|induction of
					apoptosis\|integral to
					membrane\|protein
					heterodimerization
					activity\|regulation of apoptosis
miR-125b	NM_013449	BAZ2A	bromodomain adjacent to	P + T	DNA binding\|chromatin
			zinc finger domain, 2A		remodeling\|nucleolus organizer
					complex\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					regulator activity
miR-125b	NM_004634	BRPF1	bromodomain and PHD	M + P + T	DNA
			finger containing, 1		binding\|nucleus\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|zinc ion
					binding
miR-125b	NM_003458	BSN	bassoon (presynaptic	P + T	cytoskeleton\|metal ion
			cytomatrix protein)		binding\|nucleus\|structural constituent
					of cytoskeleton\|synapse\|synaptic
					transmission\|synaptosome
miR-125b	NM_018108	C14orf130	hypothetical protein	P + T	ubiquitin cycle\|ubiquitin-protein
			FLJ10483		ligase activity
miR-125b	AA025877	C20orf136	chromosome 20 open	P + T
			reading frame 136
miR-125b	AB054985	CACNB1	calcium channel, voltage-	M + P + T	calcium ion transport\|ion
			dependent, beta 1 subunit		transport\|membrane fraction\|muscle
					contraction\|voltage-gated calcium
					channel activity\|voltage-gated
					calcium channel complex
miR-125b	NM_001224	CASP2	caspase	2, apoptosis-related	P + T	anti-apoptosis\|apoptotic
			cysteine protease (neural		program\|caspase activity\|caspase
			precursor cell expressed,		activity\|caspase activity\|cysteine-
			developmentally down-		type peptidase activity\|enzyme
			regulated 2)		binding\|intracellular\|protein
					binding\|proteolysis and
					peptidolysis\|proteolysis and
					peptidolysis\|regulation of apoptosis
miR-125b	NM_001755	CBFB	core-binding factor, beta	M + P + T	RNA polymerase II transcription
			subunit		factor activity\|nucleus\|transcription
					coactivator activity\|transcription
					factor activity\|transcription from
					RNA polymerase II promoter
miR-125b	AV648364	CBX7	ESTs, Highly similar to	P + T	chromatin\|chromatin assembly or
			potassium voltage-gated		disassembly\|chromatin
			channel, Isk-related		binding\|chromatin
			subfamily, gene 4;		modification\|nucleus\|regulation of
			potassium voltage-gated		transcription, DNA-
			channel-like protein, Isk-		dependent\|transcription
			related subfamily
			[Homo sapiens]
			[H. sapiens]
miR-125b	NM_001408	CELSR2	cadherin, EGF LAG seven-	M + P + T	G-protein coupled receptor
			pass G-type receptor 2		activity\|calcium ion binding\|cell
			(flamingo homolog,		adhesion\|development\|homophilic
			Drosophila)		cell adhesion\|integral to
					membrane\|membrane\|neuropeptide
					signaling pathway\|receptor
					activity\|signal transduction\|structural
					molecule activity
miR-125b	NM_015955	CGI-27	C21orf19-like protein	P + T
miR-125b	AF263462	CGN	cingulin	P + T	actin binding\|biological_process
					unknown\|motor
					activity\|myosin\|protein binding\|tight
					junction
miR-125b	AF064491	CLIM2	LIM domain binding 1	P + T	LIM domain
					binding\|development\|development\|negative
					regulation of transcription,
					DNA-
					dependent\|nucleus\|transcription
					cofactor activity\|transcriptional
					repressor activity
miR-125b	AU152178	CMG2	capillary morphogenesis	P + T	integral to membrane\|receptor
			protein
2		activity
miR-125b	NM_004073	CNK	cytokine-inducible kinase	P + T	ATP binding\|protein amino acid
					phosphorylation\|protein
					binding\|protein serine/threonine
					kinase activity\|regulation of cell
					cycle\|transferase activity
miR-125b	NM_020348	CNNM1	cyclin M1	M + P + T	fatty acid biosynthesis
miR-125b	NM_022730	COPS7B	COP9 constitutive	M + P + T	signalosome complex
			photomorphogenic homolog
			subunit 7B (Arabidopsis)
miR-125b	NM_003389	CORO2A	coronin, actin binding	P + T	actin binding\|glutamate-ammonia
			protein, 2A		ligase activity\|glutamine
					biosynthesis\|intracellular signaling
					cascade\|nitrogen compound
					metabolism\|protein binding
miR-125b	BF939649	CORO2B	coronin, actin binding	P + T	actin binding\|actin cytoskeleton\|actin
			protein, 2B		cytoskeleton organization and
					biogenesis\|membrane
miR-125b	NM_007007	CPSF6	cleavage and	P + T	RNA binding\|mRNA
			polyadenylation specific		processing\|nucleic acid
			factor
6, 68 kDa		binding\|nucleotide binding\|nucleus
miR-125b	NM_004386	CSPG3	chondroitin sulfate	P + T	calcium ion binding\|cell
			proteoglycan 3 (neurocan)		adhesion\|cell motility\|hyaluronic acid
					binding\|sugar binding
miR-125b	NM_004393	DAG1	dystroglycan 1 (dystrophin-	M + P + T	actin cytoskeleton\|calcium ion
			associated glycoprotein 1)		binding\|extracellular matrix (sensu
					Metazoa)\|integral to plasma
					membrane\|laminin receptor
					activity\|membrane fraction\|muscle
					contraction\|plasma
					membrane\|protein binding\|protein
					complex assembly
miR-125b	NM_014764	DAZAP2	DAZ associated protein 2	P + T
miR-125b	NM_030927	DC-TM4F2	tetraspanin similar to	P + T	integral to membrane
			TM4SF9
miR-125b	NM_004082	DCTN1	dynactin 1 (p150, glued	M + P + T	cytoplasm\|cytoskeleton\|dynein
			homolog, Drosophila)		complex\|mitosis\|motor
					activity\|neurogenesis
miR-125b	NM_030621	DICER1	Dicer1, Dcr-1 homolog	P + T	ATP binding\|ATP-dependent
			(Drosophila)		helicase activity\|RNA interference,
					targeting of mRNA for
					destruction\|RNA processing\|double-
					stranded RNA binding\|endonuclease
					activity\|hydrolase
					activity\|intracellular\|ribonuclease III
					activity
miR-125b	U53506	DIO2	deiodinase, iodothyronine,	P + T	integral to
			type II		membrane\|membrane\|selenium
					binding\|selenocysteine
					incorporation\|thyroid hormone
					generation\|thyroxine 5′-deiodinase
					activity\|thyroxine 5′-deiodinase
					activity
miR-125b	AL136139	dJ761I2.1		P + T
miR-125b	AL357503	dJ899C14.1	Q9H4T4 like	P + T
miR-125b	AL117482	DKFZP434C131	DKFZP434C131 protein	P + T	ATP binding\|protein amino acid
					phosphorylation\|protein
					serine/threonine kinase
					activity\|protein-tyrosine kinase
					activity\|transferase activity
miR-125b	AK023580	DKFZP434H0820	hypothetical protein	P + T
			DKFZp434H0820
miR-125b	T16388	DKFZp564A176	hypothetical protein	P + T	development\|integral to
			DKFZp564A176		membrane\|membrane\|receptor
					activity\|semaphorin receptor activity
miR-125b	AL137517	DKFZp564O1278	hypothetical protein	P + T	integral to membrane
			DKFZp564O1278
miR-125b	BE781961	DKFZp762A2013	hypothetical protein	P + T	electron transport\|electron
			DKFZp762A2013		transporter activity
miR-125b	AB036931	DLL4	delta-like 4 (Drosophila)	M + P + T	Notch binding\|Notch signaling
					pathway\|cell
					differentiation\|circulation\|integral to
					membrane\|membrane\|signal
					transduction
miR-125b	NM_012266	DNAJB5	DnaJ (Hsp40) homolog,	P + T	heat shock protein binding\|protein
			subfamily B, member 5		folding\|response to unfolded
					protein\|unfolded protein binding
miR-125b	NM_005740	DNAL4	dynein, axonemal, light	P + T	ATPase activity, coupled\|axonemal
			polypeptide 4		dynein complex\|microtubule motor
					activity\|microtubule-based
					movement
miR-125b	BF593175	DOCK3	dedicator of cyto-kinesis 3	P + T	GTP binding\|GTPase
					binding\|guanyl-nucleotide exchange
					factor activity
miR-125b	NM_006426	DPYSL4	dihydropyrimidinase-like 4	P + T	hydrolase activity\|neurogenesis
miR-125b	NM_006465	DRIL2	dead ringer (Drosophila)-	P + T	DNA binding\|biological_process
			like 2 (bright and dead		unknown\|nucleus
			ringer)
miR-125b	BC005047	DUSP6	dual specificity phosphatase 6	P + T	MAP kinase phosphatase
					activity\|cytoplasm\|hydrolase
					activity\|inactivation of
					MAPK\|protein amino acid
					dephosphorylation\|protein
					serine/threonine phosphatase
					activity\|protein tyrosine phosphatase
					activity\|regulation of cell
					cycle\|soluble fraction
miR-125b	NM_004423	DVL3	dishevelled, dsh homolog 3	P + T	development\|frizzled signaling
			(Drosophila)		pathway\|heart
					development\|intracellular\|intracellular
					signaling cascade\|kinase
					activity\|neurogenesis\|protein
					binding\|signal transducer activity
miR-125b	NM_001949	E2F3	E2F transcription factor 3	P + T	nucleus\|protein binding\|regulation of
					cell cycle\|regulation of transcription,
					DNA-
					dependent\|transcription\|transcription
					factor activity\|transcription factor
					complex\|transcription initiation from
					RNA polymerase II promoter
miR-125b	AU149385	EAF1	Homo sapiens cDNA	P + T
			FLJ13155 fis, clone
			NT2RP3003433, mRNA
			sequence
miR-125b	NM_014674	EDEM	KIAA0212 gene product	P + T	ER-associated protein
					catabolism\|GTP binding\|N-linked
					glycosylation\|calcium ion
					binding\|endoplasmic
					reticulum\|integral to endoplasmic
					reticulum membrane\|integral to
					membrane\|mannosyl-oligosaccharide
					1,2-alpha-mannosidase
					activity\|membrane\|protein
					binding\|response to unfolded protein
miR-125b	NM_001955	EDN1	endothelin 1	M + P + T	cell-cell signaling\|extracellular
					space\|hormone
					activity\|pathogenesis\|positive
					regulation of cell
					proliferation\|regulation of blood
					pressure\|regulation of
					vasoconstriction\|signal
					transduction\|soluble fraction
miR-125b	AI832074	EIF2C2	eukaryotic translation	M + P	cellular_component unknown\|protein
			initiation factor
2C, 2		biosynthesis\|translation initiation
					factor activity
miR-125b	AB044548	EIF4EBP1	eukaryotic translation	P + T	eukaryotic initiation factor 4E
			initiation factor 4E binding		binding\|negative regulation of
			protein 1		protein biosynthesis\|negative
					regulation of translational
					initiation\|regulation of translation
miR-125b	NM_020390	EIF5A2	eukaryotic translation	P + T	DNA binding\|protein
			initiation factor 5A2		biosynthesis\|translation initiation
					factor activity\|translational initiation
miR-125b	NM_004438	EPHA4	EphA4	P + T	ATP binding\|ephrin receptor
					activity\|integral to plasma
					membrane\|membrane\|protein amino
					acid phosphorylation\|receptor
					activity\|signal
					transduction\|transferase
					activity\|transmembrane receptor
					protein tyrosine kinase signaling
					pathway
miR-125b	NM_004451	ESRRA	estrogen-related receptor	P + T	nucleus\|regulation of transcription,
			alpha		DNA-dependent\|steroid
					binding\|steroid hormone receptor
					activity\|transcription\|transcription
					factor activity
miR-125b	NM_004907	ETR101	immediate early protein	P + T
miR-125b	NM_005238	ETS1	v-ets erythroblastosis virus	P + T	RNA polymerase II transcription
			E26 oncogene homolog 1		factor activity\|immune
			(avian)		response\|negative regulation of cell
					proliferation\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					factor activity\|transcription from
					RNA polymerase II promoter
miR-125b	NM_001987	ETV6	ets variant gene 6 (TEL	P + T	nucleus\|regulation of transcription,
			oncogene)		DNA-
					dependent\|transcription\|transcription
					factor activity
miR-125b	NM_022763	FAD104	FAD104	P + T
miR-125b	AF308300	FAPP2	phosphoinositol 4-phosphate	P + T
			adaptor protein-2
miR-125b	NM_022976	FGFR2	fibroblast growth factor	M + P + T	ATP binding\|cell growth\|fibroblast
			receptor 2 (bacteria-		growth factor receptor
			expressed kinase,		activity\|heparin binding\|integral to
			keratinocyte growth factor		membrane\|membrane\|protein amino
			receptor, craniofacial		acid phosphorylation\|protein amino
			dysostosis
1, Crouzon		acid phosphorylation\|protein
			syndrome, Pfeiffer		serine/threonine kinase
			syndrome, Jackson-Weiss		activity\|protein-tyrosine kinase
			syndrome)		activity\|protein-tyrosine kinase
					activity\|receptor activity\|transferase
					activity
miR-125b	NM_004470	FKBP2	FK506 binding protein 2,	P + T	FK506 binding\|endoplasmic
			13 kDa		reticulum\|isomerase
					activity\|peptidyl-prolyl cis-trans
					isomerase activity\|protein folding
miR-125b	AL160175	FKHL18	forkhead-like 18	P + T
			(Drosophila)
miR-125b	BF515132	FLJ00024	hypothetical protein	P + T
			FLJ00024
miR-125b	BC002945	FLJ10101	hypothetical protein	M + P	GTP binding\|protein transport\|small
			FLJ10101		GTPase mediated signal transduction
miR-125b	NM_018243	FLJ10849	hypothetical protein	P + T	GTP binding\|cell cycle\|cytokinesis
			FLJ10849
miR-125b	NM_019084	FLJ10895	hypothetical protein	P + T	nucleus\|regulation of cell cycle
			FLJ10895
miR-125b	NM_018320	FLJ11099	hypothetical protein	P + T	protein ubiquitination\|ubiquitin
			FLJ11099		ligase complex\|ubiquitin-protein
					ligase activity\|zinc ion binding
miR-125b	NM_018375	FLJ11274	hypothetical protein	M + P + T	membrane\|metal ion transport\|metal
			FLJ11274		ion transporter activity
miR-125b	NM_024954	FLJ11807	hypothetical protein	P + T	protein modification
			FLJ11807
miR-125b	BF434995	FLJ14708	hypothetical protein	P + T
			FLJ14708
miR-125b	NM_018992	FLJ20040	hypothetical protein	P + T	membrane\|potassium ion
			FLJ20040		transport\|protein binding\|voltage-
					gated potassium channel
					activity\|voltage-gated potassium
					channel complex
miR-125b	NM_017911	FLJ20635	hypothetical protein	P + T
			FLJ20635
miR-125b	NM_017936	FLJ20707	hypothetical protein	M + P + T	ATP synthesis coupled proton
			FLJ20707		transport\|cytoplasm\|hydrogen-
					transporting ATP synthase activity,
					rotational mechanism\|hydrogen-
					transporting ATPase activity,
					rotational
					mechanism\|membrane\|phosphate
					transport\|proton-transporting two-
					sector ATPase complex
miR-125b	NM_024789	FLJ22529	hypothetical protein	P + T
			FLJ22529
miR-125b	AA721230	FLJ25604	hypothetical protein	P + T	guanyl-nucleotide exchange factor
			FLJ25604		activity\|small GTPase mediated
					signal transduction
miR-125b	AI677701	FLJ30829	hypothetical protein	P + T	nucleic acid binding\|nucleotide
			FLJ30829		binding
miR-125b	NM_004475	FLOT2	flotillin 2	M + P + T	cell adhesion\|epidermis
					development\|flotillin
					complex\|integral to
					membrane\|plasma membrane\|protein
					binding
miR-125b	AA830884	FMR1	fragile X mental retardation 1	M + T	mRNA binding\|mRNA
					processing\|mRNA-nucleus
					export\|nucleoplasm\|polysome\|ribo-
					some\|soluble fraction\|transport
miR-125b	AF305083	FUT4	fucosyltransferase 4 (alpha	P + T	Golgi apparatus\|L-fucose
			(1,3) fucosyltransferase,		catabolism\|alpha(1,3)-
			myeloid-specific)		fucosyltransferase
					activity\|carbohydrate
					metabolism\|integral to
					membrane\|membrane\|membrane
					fraction\|protein amino acid
					glycosylation\|transferase activity,
					transferring glycosyl groups
miR-125b	X92762	G4.5	tafazzin (cardiomyopathy,	M + P + T	acyltransferase activity\|heart
			dilated 3A (X-linked);		development\|integral to
			endocardial fibroelastosis 2;		membrane\|metabolism\|muscle
			Barth syndrome)		contraction\|muscle development
miR-125b	NM_012296	GAB2	GRB2-associated binding	P + T
			protein
2
miR-125b	NM_015044	GGA2	golgi associated, gamma	M + T	ADP-ribosylation factor
			adaptin ear containing, ARF		binding\|Golgi stack\|Golgi trans face\|
			binding protein 2		clathrin coat of trans-Golgi
					network vesicle\|intra-Golgi
					transport\|intracellular protein
					transport\|intracellular protein
					transport\|membrane\|protein complex
					assembly\|protein transporter activity
miR-125b	AL049709	GGTL3	gamma-glutamyltransferase-	M + P + T
			like 3
miR-125b	NM_000165	GJA1	gap junction protein, alpha	P + T	cell-cell signaling\|connexon channel
			1, 43 kDa (connexin 43)		activity\|connexon complex\|gap
					junction assembly\|heart
					development\|integral to plasma
					membrane\|ion transporter
					activity\|muscle
					contraction\|perception of
					sound\|positive regulation of I-
					kappaB kinase/NF-kappaB
					cascade\|protein binding\|signal
					transducer activity\|transport
miR-125b	NM_014905	GLS	glutaminase	P + T	glutaminase activity\|glutamine
					catabolism\|hydrolase
					activity\|mitochondrion
miR-125b	NM_005113	GOLGA5	golgi autoantigen, golgin	P + T	ATP binding\|Golgi membrane\|cell
			subfamily a, 5		surface receptor linked signal
					transduction\|integral to plasma
					membrane\|protein amino acid
					phosphorylation\|protein-tyrosine
					kinase activity
miR-125b	NM_001448	GPC4	glypican 4	M + P + T	cell proliferation\|extracellular matrix
					(sensu Metazoa)\|integral to plasma
					membrane\|membrane\|morphogenesis
miR-125b	NM_005296	GPR23	G protein-coupled receptor	M + T	G-protein coupled receptor protein
			23		signaling pathway\|integral to plasma
					membrane\|purinergic nucleotide
					receptor activity, G-protein
					coupled\|receptor activity\|rhodopsin-
					like receptor activity\|signal
					transduction
miR-125b	U66065	GRB10	growth factor receptor-	M + T	SH3/SH2 adaptor activity\|cell-cell
			bound protein 10		signaling\|cytoplasm\|insulin receptor
					signaling pathway\|intracellular
					signaling cascade\|plasma membrane
miR-125b	NM_021643	GS3955	GS3955 protein	P + T	ATP binding\|protein amino acid
					phosphorylation\|protein kinase
					activity\|transferase activity
miR-125b	NM_019096	GTPBP2	GTP binding protein 2	M + T	GTP binding\|GTPase activity\|protein
					biosynthesis\|small GTPase mediated
					signal transduction
miR-125b	U78181	hBNaC2	amiloride-sensitive cation	P + T	amiloride-sensitive sodium channel
			channel
2, neuronal		activity\|integral to plasma
					membrane\|ion channel activity\|ion
					transport\|membrane\|response to
					pH\|signal transduction\|sodium ion
					transport
miR-125b	NM_005477	HCN4	hyperpolarization activated	P + T	3′,5′-cAMP binding\|cation channel
			cyclic nucleotide-gated		activity\|cation
			potassium channel 4		transport\|circulation\|integral to
					plasma
					membrane\|membrane\|membrane
					fraction\|muscle
					contraction\|nucleotide
					binding\|potassium ion
					transport\|sodium ion
					transport\|voltage-gated potassium
					channel activity
miR-125b	NM_002112	HDC	histidine decarboxylase	P + T	amino acid
					metabolism\|catecholamine
					biosynthesis\|histidine decarboxylase
					activity\|histidine metabolism\|lyase
					activity
miR-125b	U64317	HEF1	enhancer of filamentation 1	P + T	actin filament bundle formation\|cell
			(cas-like docking; Crk-		adhesion\|cytokinesis\|cytoplasm\|cyto-
			associated substrate related)		skeleton\|cytoskeleton organization
					and biogenesis\|integrin-mediated
					signaling
					pathway\|mitosis\|nucleus\|protein
					binding\|regulation of cell
					cycle\|regulation of cell growth\|signal
					transduction\|spindle
miR-125b	L38487	hERRa	estrogen-related receptor	P + T	nucleus\|regulation of transcription,
			alpha		DNA-dependent\|steroid
					binding\|steroid hormone receptor
					activity\|transcription\|transcription
					factor activity
miR-125b	AB028943	HIC2	hypermethylated in cancer 2	P + T	DNA binding\|negative regulation of
					transcription, DNA-
					dependent\|nucleus\|protein C-
					terminus binding\|transcription\|zinc
					ion binding
miR-125b	AL023584	HIVEP2	human immunodeficiency	P + T
			virus type I enhancer
			binding protein
2
miR-125b	AL023584	HIVEP2	human immunodeficiency	P + T
			virus type I enhancer
			binding protein
2
miR-125b	NM_005342	HMGB3	high-mobility group box 3	P + T	DNA bending activity\|DNA
					binding\|chromatin\|development\|nu-
					cleus\|regulation of transcription, DNA-
					dependent
miR-125b	AL031295	HMGCL; HL	lysophospholipase II	M + P + T
miR-125b	NM_004503	HOXC6	homeo box C6	P + T	development\|development\|nucleus\|regulation
					of transcription from RNA
					polymerase II promoter\|regulation of
					transcription, DNA-
					dependent\|transcription corepressor
					activity\|transcription factor activity
miR-125b	AA844682	HRD1	HRD1 protein	P + T	protein ubiquitination\|ubiquitin
					ligase complex\|ubiquitin-protein
					ligase activity\|zinc ion binding
miR-125b	AL136667	HSPC039	HSPC039 protein	P + T	integral to membrane
miR-125b	AF245044	HT023	hypothetical protein HT023	P + T
miR-125b	U13022	Ich-1	caspase 2, apoptosis-related	P + T	anti-apoptosis\|apoptotic
			cysteine protease (neural		program\|caspase activity\|caspase
			precursor cell expressed,		activity\|caspase activity\|cysteine-
			developmentally down-		type peptidase activity\|enzyme
			regulated 2)		binding\|intracellular\|protein
					binding\|proteolysis and
					peptidolysis\|proteolysis and
					peptidolysis\|regulation of apoptosis
miR-125b	NM_004513	IL16	interleukin 16 (lymphocyte	M + P + T	chemotaxis\|cytokine
			chemoattractant factor)		activity\|extracellular space\|immune
					response\|protein binding\|sensory
					perception
miR-125b	NM_002460	IRF4	interferon regulatory factor 4	P + T	RNA polymerase II transcription
					factor activity\|T-cell activation\|T-cell
					activation\|nucleus\|nucleus\|nucleus\|positive
					regulation of interleukin-10
					biosynthesis\|positive regulation of
					interleukin-10 biosynthesis\|positive
					regulation of interleukin-13
					biosynthesis\|positive regulation of
					interleukin-13 biosynthesis\|positive
					regulation of interleukin-2
					biosynthesis\|positive regulation of
					interleukin-2 biosynthesis\|positive
					regulation of interleukin-4
					biosynthesis\|positive regulation of
					interleukin-4 biosynthesis\|positive
					regulation of transcription\|positive
					regulation of transcription\|regulation
					of T-helper cell
					differentiation\|regulation of T-helper
					cell differentiation\|regulation of
					transcription, DNA-
					dependent\|regulation of transcription,
					DNA-
					dependent\|transcription\|transcription
					factor activity\|transcription factor
					activity\|transcription factor
					binding\|transcription factor
					binding\|transcriptional activator
					activity\|transcriptional activator
					activity
miR-125b	NM_002207	ITGA9	integrin, alpha 9	P + T	cell-matrix adhesion\|integral to
					membrane\|integrin complex\|integrin-
					mediated signaling pathway\|protein
					binding\|receptor activity
miR-125b	NM_000212	ITGB3	integrin, beta 3 (platelet	P + T	blood coagulation\|cell-matrix
			glycoprotein IIIa, antigen		adhesion\|integrin complex\|integrin-
			CD61)		mediated signaling pathway\|protein
					binding\|receptor activity
miR-125b	NM_021991	JUP	junction plakoglobin	P + T	cell adhesion\|cell
					adhesion\|cytoplasm\|cytoskeletal
					protein
					binding\|cytoskeleton\|cytoskeleton\|membrane
					fraction\|mitotic chromosome
					condensation\|protein binding\|soluble
					fraction\|structural molecule activity
miR-125b	AF032897	KCNH7	potassium voltage-gated	P + T	cation transport\|integral to
			channel, subfamily H (eag-		membrane\|membrane\|potassium ion
			related), member 7		transport\|regulation of transcription,
					DNA-dependent\|signal transducer
					activity\|signal transduction\|voltage-
					gated potassium channel activity
miR-125b	NM_002252	KCNS3	potassium voltage-gated	M + P + T	cation transport\|delayed rectifier
			channel, delayed-rectifier,		potassium channel
			subfamily S, member 3		activity\|membrane\|membrane
					fraction\|potassium channel regulator
					activity\|potassium ion
					transport\|protein binding\|voltage-
					gated potassium channel complex
miR-125b	NM_014735	KIAA0215	KIAA0215 gene product	P + T	DNA binding\|regulation of
					transcription, DNA-dependent
miR-125b	NM_015288	KIAA0239	KIAA0239 protein	P + T	DNA binding\|regulation of
					transcription, DNA-dependent
miR-125b	D87469	KIAA0279	cadherin, EGF LAG seven-	M + P + T	G-protein coupled receptor
			pass G-type receptor 2		activity\|calcium ion binding\|cell
			(flamingo homolog,		adhesion\|development\|homophilic
			Drosophila)		cell adhesion\|integral to
					membrane\|membrane\|neuropeptide
					signaling pathway\|receptor
					activity\|signal transduction\|structural
					molecule activity
miR-125b	AB002356	KIAA0358	MAP-kinase activating	P + T	cell surface receptor linked signal
			death domain		transduction\|cytoplasm\|death
					receptor binding\|kinase
					activity\|plasma membrane\|protein
					kinase activator activity
miR-125b	NM_014871	KIAA0710	KIAA0710 gene product	P + T	cysteine-type endopeptidase
					activity\|exonuclease
					activity\|nucleus\|ubiquitin
					cycle\|ubiquitin thiolesterase
					activity\|ubiquitin-dependent protein
					catabolism
miR-125b	AB018333	KIAA0790	KIAA0790 protein	P + T	cell cycle\|negative regulation of cell
					cycle
miR-125b	NM_014912	KIAA0940	KIAA0940 protein	P + T	nucleic acid binding
miR-125b	AB028957	KIAA1034	KIAA1034 protein	P + T	DNA binding\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription factor
					activity
miR-125b	NM_014901	KIAA1100	KIAA1100 protein	M + P + T	protein ubiquitination\|ubiquitin
					ligase complex\|ubiquitin-protein
					ligase activity\|zinc ion binding
miR-125b	AB033016	KIAA1190	hypothetical protein	P + T	DNA binding\|nucleic acid
			KIAA1190		binding\|nucleus\|protein
					binding\|regulation of transcription,
					DNA-dependent\|zinc ion binding
miR-125b	AA056548	KIAA1268	KIAA1268 protein	P + T	NAD+ ADP-ribosyltransferase
					activity\|nucleus\|protein amino acid
					ADP-ribosylation
miR-125b	BE670098	KIAA1594	KIAA1594 protein	M + P + T	cysteine-type endopeptidase
					activity\|ubiquitin cycle\|ubiquitin
					thiolesterase activity\|ubiquitin-
					dependent protein catabolism
miR-125b	AU157109	KIAA1598	KIAA1598 protein	P + T
miR-125b	AA772278	KIAA1673	KIAA1673	P + T
miR-125b	NM_015995	KLF13	Kruppel-like factor 13	P + T	DNA binding\|RNA polymerase II
					transcription factor
					activity\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					from RNA polymerase II
					promoter\|zinc ion binding
miR-125b	NM_016531	KLF3	Kruppel-like factor 3 (basic)	P + T	development\|negative regulation of
					transcription from RNA polymerase
					II promoter\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					factor activity\|zinc ion binding
miR-125b	BE892574	LACTB	lactamase, beta	P + T	hydrolase activity\|integral to
					membrane\|response to antibiotic
miR-125b	BE566136	LBP-32	LBP protein 32	P + T
miR-125b	NM_024090	LCE	long-chain fatty-acyl	P + T	integral to membrane
			elongase
miR-125b	NM_003893	LDB1	LIM domain binding 1	P + T	LIM domain
					binding\|development\|development\|negative
					regulation of transcription, DNA-
					dependent\|nucleus\|transcription
					cofactor activity\|transcriptional
					repressor activity
miR-125b	U94354	LFNG	lunatic fringe homolog	M + T	Golgi
			(Drosophila)		apparatus\|development\|extracellular
					region\|integral to
					membrane\|membrane\|organo-
					genesis\|transferase activity,
					transferring glycosyl groups
miR-125b	NM_002310	LIFR	leukemia inhibitory factor	M + P + T	cell surface receptor linked signal
			receptor		transduction\|integral to plasma
					membrane\|leukemia inhibitory factor
					receptor activity\|membrane\|receptor
					activity
miR-125b	NM_016339	Link-GEFII	Link guanine nucleotide	P + T	G-protein coupled receptor protein
			exchange factor II		signaling pathway\|guanyl-nucleotide
					exchange factor activity\|membrane
					fraction\|neurogenesis\|small GTPase
					mediated signal transduction
miR-125b	NM_005575	LNPEP	leucyl/cystinyl	P + T	aminopeptidase activity\|cell-cell
			aminopeptidase		signaling\|integral to plasma
					membrane\|membrane alanyl
					aminopeptidase
					activity\|metallopeptidase
					activity\|plasma
					membrane\|pregnancy\|proteolysis and
					peptidolysis\|zinc ion binding
miR-125b	AL031186	LOC129080	putative emu1	P + T
miR-125b	AI884701	LOC221002	CG4853 gene product	M + P	guanyl-nucleotide exchange factor
					activity\|small GTPase mediated
					signal transduction
miR-125b	AI953847	LOC255488	Homo sapiens mRNA full	P + T	electron transport\|electron
			length insert cDNA clone		transporter activity\|integral to
			EUROIMAGE 186647,		membrane\|iron ion binding\|ligase
			mRNA sequence		activity\|protein binding\|protein
					ubiquitination during ubiquitin-
					dependent protein
					catabolism\|ubiquitin ligase
					complex\|ubiquitin-protein ligase
					activity\|zinc ion binding
miR-125b	NM_015899	LOC51054	putative glycolipid transfer	P + T
			protein
miR-125b	AA209239	LOC57406	lipase protein	P + T	aromatic compound
					metabolism\|hydrolase
					activity\|response to toxin\|xenobiotic
					metabolism
miR-125b	NM_005576	LOXL1	lysyl oxidase-like 1	M + P + T	copper ion binding\|electron
					transporter activity\|extracellular
					region\|oxidoreductase
					activity\|protein modification\|protein-
					lysine 6-oxidase activity
miR-125b	AA584297	LRP4	low density lipoprotein	M + T	calcium ion
			receptor-related protein 4		binding\|endocytosis\|integral to
					membrane\|membrane\|receptor
					activity
miR-125b	NM_007260	LYPLA2	lysophospholipase II	M + P + T	fatty acid metabolism\|hydrolase
					activity\|lipid metabolism
miR-125b	NM_004901	LYSAL1	lysosomal apyrase-like 1	P + T	Golgi apparatus\|UDP
					catabolism\|apyrase activity\|hydrolase
					activity\|integral to Golgi
					membrane\|integral to
					membrane\|lysosome\|magnesium ion
					binding\|nucleobase, nucleoside,
					nucleotide and nucleic acid
					metabolism\|uridine-diphosphatase
					activity\|vacuolar membrane
miR-125b	NM_002355	M6PR	mannose-6-phosphate	M + P + T	endosome to lysosome
			receptor (cation dependent)		transport\|integral to plasma
					membrane\|lysosome\|receptor
					mediated endocytosis\|transmembrane
					receptor activity\|transport\|transporter
					activity
miR-125b	AB002356	MADD	MAP-kinase activating	P + T	cell surface receptor linked signal
			death domain		transduction\|cytoplasm\|death
					receptor binding\|kinase
					activity\|plasma membrane\|protein
					kinase activator activity
miR-125b	NM_016219	MAN1B1	mannosidase, alpha, class	P + T	N-linked glycosylation\|N-linked
			1B, member 1		glycosylation\|calcium ion
					binding\|calcium ion
					binding\|carbohydrate
					metabolism\|endoplasmic
					reticulum\|hydrolase activity, acting
					on glycosyl bonds\|integral to
					membrane\|mannosyl-oligosaccharide
					1,2-alpha-mannosidase
					activity\|mannosyl-oligosaccharide
					1,2-alpha-mannosidase
					activity\|membrane\|membrane
					fraction\|oligosaccharide metabolism
miR-125b	NM_002446	MAP3K10	mitogen-activated protein	P + T	ATP binding\|JUN kinase kinase
			kinase kinase kinase 10		kinase activity\|activation of
					JNK\|autophosphorylation\|induction
					of apoptosis\|protein
					homodimerization activity\|protein
					serine/threonine kinase
					activity\|protein-tyrosine kinase
					activity\|signal
					transduction\|transferase activity
miR-125b	NM_002419	MAP3K11	mitogen-activated protein	M + P + T	ATP binding\|G1 phase of mitotic cell
			kinase kinase kinase 11		cycle\|JUN kinase kinase kinase
					activity\|activation of
					JNK\|autophosphorylation\|cell
					proliferation\|centrosome\|microtubule\|
					microtubule-based process\|protein
					homodimerization activity\|protein
					oligomerization\|protein
					serine/threonine kinase
					activity\|protein-tyrosine kinase
					activity\|transferase activity
miR-125b	Z25432	MAPK14	mitogen-activated protein	P + T	ATP binding\|MAP kinase
			kinase
14		activity\|MAP kinase kinase
					activity\|MP kinase
					activity\|antimicrobial humoral
					response (sensu Vertebrata)\|cell
					motility\|cell surface receptor linked
					signal
					transduction\|chemotaxis\|cytoplasm\|nu-
					cleus\|protein amino acid
					phosphorylation\|protein kinase
					cascade\|protein serine/threonine
					kinase activity\|protein-tyrosine
					kinase activity\|response to
					stress\|transferase activity
miR-125b	NM_018650	MARK1	MAP/microtubule affinity-	P + T	ATP
			regulating kinase
1		binding\|cytoplasm\|cytoskeleton\|cytoskeleton
					organization and
					biogenesis\|magnesium ion
					binding\|microtubule
					cytoskeleton\|protein amino acid
					phosphorylation\|protein amino acid
					phosphorylation\|protein kinase
					cascade\|protein serine/threonine
					kinase activity\|protein
					serine/threonine kinase
					activity\|transferase activity
miR-125b	NM_001879	MASP1	mannan-binding lectin	P + T	calcium ion binding\|chymotrypsin
			serine protease 1 (C4/C2		activity\|complement
			activating component of Ra-		activation\|complement activation,
			reactive factor)		classical pathway\|extracellular
					region\|immune response\|peptidase
					activity\|proteolysis and
					peptidolysis\|trypsin activity
miR-125b	NM_005911	MAT2A	methionine	P + T	ATP binding\|magnesium ion
			adenosyltransferase II, alpha		binding\|methionine
					adenosyltransferase activity\|one-
					carbon compound
					metabolism\|transferase activity
miR-125b	NM_005920	MEF2D	MADS box transcription	P + T	muscle
			enhancer factor
2,		development\|nucleus\|regulation of
			polypeptide D (myocyte		transcription, DNA-
			enhancer factor 2D)		dependent\|transcription\|transcription
					coactivator activity\|transcription
					factor activity\|transcription from
					RNA polymerase II promoter
miR-125b	NM_020149	MEIS2	Meis1, myeloid ecotropic	M + P	negative regulation of transcription
			viral integration site 1		from RNA polymerase II
			homolog 2 (mouse)		promoter\|nucleus\|regulation of
					transcription, DNA-
					dependent\|specific RNA polymerase
					II transcription factor
					activity\|transcription corepressor
					activity\|transcription factor
					activity\|transcription factor activity
miR-125b	NM_017927	MFN1	mitofusin 1	P + T	GTP binding\|GTPase
					activity\|hydrolase activity\|integral to
					membrane\|mitochondrial
					fusion\|mitochondrial outer
					membrane\|mitochondrion
miR-125b	AI139252	MGC16063	ribosomal protein L35a	P + T	JAK-STAT cascade\|acute-phase
					response\|calcium ion binding\|cell
					motility\|cytoplasm\|hematopoietin/interferon-
					class (D200-domain)
					cytokine receptor signal transducer
					activity\|intracellular signaling
					cascade\|negative regulation of
					transcription from RNA polymerase II
					promoter\|neurogenesis\|nucleus\|nu-
					cleus\|regulation of transcription, DNA-
					dependent\|signal transducer
					activity\|transcription\|transcription
					factor activity\|transcription factor
					activity
miR-125b	AI862120	MGC21981	hypothetical protein	P + T	membrane
			MGC21981
miR-125b	AL515061	MGC24302	hypothetical protein	P + T
			MGC24302
miR-125b	BE618656	MGC2541	similar to RIKEN cDNA	M + P + T
			2610030J16 gene
miR-125b	BC005842	MGC2705	hypothetical protein	P + T
			MGC2705
miR-125b	NM_024293	MGC3035	hypothetical protein	M + P
			MGC3035
miR-125b	NM_017572	MKNK2	MAP kinase-interacting	P + T	ATP binding\|ATP binding\|cell
			serine/threonine kinase 2		surface receptor linked signal
					transduction\|protein amino acid
					phosphorylation\|protein amino acid
					phosphorylation\|protein kinase
					cascade\|protein serine/threonine
					kinase activity\|protein
					serine/threonine kinase
					activity\|protein-tyrosine kinase
					activity\|regulation of
					translation\|response to
					stress\|transferase activity
miR-125b	NM_005439	MLF2	myeloid leukemia factor 2	P + T	defense response\|nucleus
miR-125b	NM_007359	MLN51	MLN51 protein	P + T	mRNA processing\|mRNA-nucleus
					export\|molecular_function
					unknown\|nucleus\|transport
miR-125b	NM_002442	MSI1	musashi homolog 1	M + P + T	RNA
			(Drosophila)		binding\|neurogenesis\|nucleotide
					binding\|nucleus
miR-125b	NM_021090	MTMR3	myotubularin related protein 3	M + P + T	cytoplasm\|hydrolase activity\|inositol
					or phosphatidylinositol phosphatase
					activity\|membrane\|membrane
					fraction\|phospholipid
					dephosphorylation\|protein amino
					acid dephosphorylation\|protein
					serine/threonine phosphatase
					activity\|protein tyrosine phosphatase
					activity\|protein
					tyrosine/serine/threonine
					phosphatase activity\|zinc ion binding
miR-125b	AK024501	MXD4	MAX dimerization protein 4	M + P + T	DNA binding\|negative regulation of
					cell proliferation\|negative regulation
					of transcription from RNA
					polymerase II
					promoter\|nucleus\|protein
					binding\|regulation of transcription,
					DNA-
					dependent\|transcription\|transcription
					corepressor activity
miR-125b	AB020642	MYT1	myelin transcription factor 1	M + P + T	nucleus\|regulation of transcription,
					DNA-
					dependent\|transcription\|transcription
					factor activity\|zinc ion binding
miR-125b	NM_004540	NCAM2	neural cell adhesion	P + T	cell adhesion\|integral to
			molecule 2		membrane\|membrane\|neuron
					adhesion\|plasma membrane\|protein
					binding
miR-125b	NM_012338	NET-2	transmembrane 4	P + T	integral to membrane\|membrane
			superfamily member		fraction
			tetraspan NET-2
miR-125b	U84246	NEU1	sialidase 1 (lysosomal	P + T	carbohydrate metabolism\|exo-alpha-
			sialidase)		sialidase activity\|hydrolase activity,
					acting on glycosyl bonds\|lysosome
miR-125b	AI824012	NRIP1	nuclear receptor interacting	P + T	nucleus\|regulation of transcription,
			protein 1		DNA-
					dependent\|transcription\|transcription
					coactivator activity
miR-125b	D81048	NRM	nurim (nuclear envelope	P + T
			membrane protein)
miR-125b	BC001794	NUMBL	numb homolog	P + T	neurogenesis
			(Drosophila)-like
miR-125b	AB020713	NUP210	nucleoporin 210	P + T	development\|nucleus
miR-125b	NM_002537	OAZ2	ornithine decarboxylase	M + P + T	ornithine decarboxylase inhibitor
			antizyme
2		activity\|polyamine metabolism
miR-125b	NM_024586	OSBPL9	oxysterol binding protein-	P + T	lipid transport\|steroid metabolism
			like 9
miR-125b	U64661	PABP	ESTs, Highly similar to	P + T
			PAB1_HUMAN
			Polyadenylate-binding
			protein 1 (Poly(A)-binding
			protein 1) (PABP 1)
			(PABP1) [H. sapiens]
miR-125b	AK000003	PCQAP	PC2 (positive cofactor 2,	P + T
			multiprotein complex)
			glutamine/Q-rich-associated
			protein
miR-125b	NM_004716	PCSK7	proprotein convertase	M + P + T	integral to Golgi membrane\|integral
			subtilisin/kexin type 7		to membrane\|peptidase
					activity\|peptidase activity\|peptide
					hormone processing\|proteolysis and
					peptidolysis\|subtilase activity
miR-125b	NM_006201	PCTK1	PCTAIRE protein kinase 1	M + P + T	ATP binding\|protein amino acid
					phosphorylation\|protein amino acid
					phosphorylation\|protein
					serine/threonine kinase
					activity\|protein serine/threonine
					kinase activity\|regulation of cell
					cycle\|transferase activity
miR-125b	NM_021213	PCTP	phosphatidylcholine transfer	M + P + T	cytosol\|lipid binding\|lipid
			protein		transport\|phosphatidylcholine
					transporter activity
miR-125b	NM_021255	PELI2	pellino homolog 2	M + P + T
			(Drosophila)
miR-125b	NM_002646	PIK3C2B	phosphoinositide-3-kinase,	P + T	inositol or phosphatidylinositol
			class
2, beta polypeptide		kinase activity\|intracellular signaling
					cascade\|microsome\|phosphatidylinositol
					3-kinase
					activity\|phosphatidylinositol-4-
					phosphate 3-kinase
					activity\|phosphoinositide 3-kinase
					complex\|plasma
					membrane\|transferase activity
miR-125b	NM_003628	PKP4	plakophilin 4	P + T	cell
					adhesion\|cytoskeleton\|intercellular
					junction\|protein binding\|structural
					molecule activity
miR-125b	NM_006718	PLAGL1	pleiomorphic adenoma	P + T	DNA binding\|cell cycle
			gene-like 1		arrest\|induction of apoptosis\|nucleic
					acid binding\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|zinc ion
					binding
miR-125b	AI457120	PPAT	phosphoribosyl	P + T	amidophosphoribosyltransferase
			pyrophosphate		activity\|glutamine
			amidotransferase		metabolism\|magnesium ion
					binding\|metabolism\|nucleoside
					metabolism\|purine base
					biosynthesis\|purine nucleotide
					biosynthesis\|transferase activity,
					transferring glycosyl groups
miR-125b	NM_002719	PPP2R5C	protein phosphatase	2,	P + T	hydrolase
			regulatory subunit B (B56),		activity\|nucleus\|phosphoprotein
			gamma isoform		phosphatase activity\|protein
					phosphatase type 2A
					complex\|protein phosphatase type
					2A complex\|protein phosphatase
					type 2A regulator activity\|protein
					phosphatase type 2A regulator
					activity\|signal transduction\|signal
					transduction
miR-125b	AL022067	PRDM1	PR domain containing 1,	P + T
			with ZNF domain
miR-125b	U23736	PRDM2	PR domain containing 2,	P + T	DNA binding\|metal ion
			with ZNF domain		binding\|nucleus\|nucleus\|regulation of
					transcription\|regulation of
					transcription, DNA-
					dependent\|transcription factor
					activity\|transcription regulator
					activity\|zinc ion binding\|zinc ion
					binding
miR-125b	AF083033	PRKRA	protein kinase, interferon-	P + T	double-stranded RNA
			inducible double stranded		binding\|enzyme activator
			RNA dependent activator		activity\|immune
					response\|intracellular\|kinase
					activity\|negative regulation of cell
					proliferation\|response to virus\|signal
					transducer activity\|signal
					transduction
miR-125b	NM_014369	PTPN18	protein tyrosine	P + T	hydrolase activity\|non-membrane
			phosphatase, non-receptor		spanning protein tyrosine
			type 18 (brain-derived)		phosphatase activity\|protein amino
					acid dephosphorylation\|protein
					amino acid
					dephosphorylation\|protein tyrosine
					phosphatase activity
miR-125b	AI762627	PTPRF	protein tyrosine	P + T	cell adhesion\|hydrolase
			phosphatase, receptor type, F		activity\|integral to membrane\|integral
					to plasma membrane\|protein amino
					acid dephosphorylation\|protein
					binding\|protein tyrosine phosphatase
					activity\|receptor
					activity\|transmembrane receptor
					protein tyrosine phosphatase
					activity\|transmembrane receptor
					protein tyrosine phosphatase
					signaling pathway
miR-125b	NM_002840	PTPRF	protein tyrosine	P + T	cell adhesion\|hydrolase
			phosphatase, receptor type, F		activity\|integral to membrane\|integral
					to plasma membrane\|protein amino
					acid dephosphorylation\|protein
					binding\|protein tyrosine phosphatase
					activity\|receptor
					activity\|transmembrane receptor
					protein tyrosine phosphatase
					activity\|transmembrane receptor
					protein tyrosine phosphatase
					signaling pathway
miR-125b	AF142419	QKI	homolog of mouse quaking	P + T
			QKI (KH domain RNA
			binding protein)
miR-125b	NM_004283	RAB3D	RAB3D, member RAS	P + T	GTP binding\|GTPase
			oncogene family		activity\|exocytosis\|hemocyte
					development\|protein transport\|small
					GTPase mediated signal transduction
miR-125b	BC002510	RAB6B	RAB6B, member RAS	P + T	GTP binding\|GTPase activity\|Golgi
			oncogene family		apparatus\|intracellular protein
					transport\|retrograde transport, Golgi
					to ER\|small GTPase mediated signal
					transduction
miR-125b	AK022662	RASAL2	RAS protein activator like 2	P + T	GTPase activator activity\|Ras
					GTPase activator activity\|signal
					transduction
miR-125b	NM_004841	RASAL2	RAS protein activator like 2	P + T	GTPase activator activity\|Ras
					GTPase activator activity\|signal
					transduction
miR-125b	NM_016090	RBM7	RNA binding motif protein 7	P + T	RNA binding\|meiosis\|nucleic acid
					binding\|nucleotide binding
miR-125b	NM_006268	REQ	requiem, apoptosis response	M + P + T	DNA binding\|apoptosis\|induction of
			zinc finger gene		apoptosis by extracellular
					signals\|nucleus\|protein
					ubiquitination\|regulation of
					transcription, DNA-
					dependent\|transcription\|ubiquitin
					ligase complex\|ubiquitin-protein
					ligase activity\|zinc ion binding
miR-125b	NM_000449	RFX5	regulatory factor X, 5	P + T	nucleus\|regulation of transcription,
			(influences HLA class II		DNA-
			expression)		dependent\|transcription\|transcription
					coactivator activity\|transcription
					factor activity\|transcription from
					RNA polymerase II promoter
miR-125b	NM_003721	RFXANK	regulatory factor X-	P + T	humoral immune
			associated ankyrin-		response\|nucleus\|regulation of
			containing protein		transcription, DNA-
					dependent\|transcription\|transcription
					coactivator activity\|transcription
					factor activity\|transcription from
					RNA polymerase II promoter
miR-125b	NM_014746	RNF144	likely ortholog of mouse	P + T	nucleus\|protein
			ubiquitin conjugating		ubiquitination\|ubiquitin ligase
			enzyme 7 interacting protein 4		complex\|ubiquitin-protein ligase
					activity\|zinc ion binding
miR-125b	NM_014771	RNF40	ring finger protein 40	M + P + T	protein ubiquitination\|ubiquitin
					ligase complex\|ubiquitin-protein
					ligase activity\|zinc ion binding
miR-125b	AL109955	RNPC1	RNA-binding region (RNP1,	P + T
			RRM) containing 1
miR-125b	AF116627	RPL29	ribosomal protein L29	M + T
miR-125b	NM_002953	RPS6KA1	ribosomal protein S6 kinase,	M + P + T	ATP binding\|protein amino acid
			90 kDa, polypeptide 1		phosphorylation\|protein
					serine/threonine kinase
					activity\|protein serine/threonine
					kinase activity\|protein-tyrosine
					kinase activity\|signal
					transduction\|transferase activity
miR-125b	NM_000332	SCA1	spinocerebellar ataxia 1	P + T	RNA binding\|cytoplasm\|nucleus
			(olivopontocerebellar ataxia
			1, autosomal dominant,
			ataxin 1)
miR-125b	NM_012429	SEC14L2	SEC14-like 2 (S. cerevisiae)	P + T	cytoplasm\|intracellular protein
					transport\|membrane\|nucleus\|phospho-
					lipid binding\|positive regulation of
					transcription, DNA-
					dependent\|protein carrier
					activity\|regulation of cholesterol
					biosynthesis\|transcription\|transcriptional
					activator
					activity\|transport\|vitamin E binding
miR-125b	NM_005065	SEL1L	sel-1 suppressor of lin-12-	P + T	catalytic activity\|integral to
			like (C. elegans)		membrane
miR-125b	NM_017789	SEMA4C	sema domain,	M + P + T	cell differentiation\|integral to
			immunoglobulin domain		membrane\|membrane\|neurogenesis\|receptor
			(Ig), transmembrane domain		activity
			(TM) and short cytoplasmic
			domain, (semaphorin) 4C
miR-125b	NM_006378	SEMA4D	sema domain,	P + T	anti-apoptosis\|cell adhesion\|cell
			immunoglobulin domain		differentiation\|immune
			(Ig), transmembrane domain		response\|integral to
			(TM) and short cytoplasmic		membrane\|membrane\|neurogenesis\|receptor
			domain, (semaphorin) 4D		activity
miR-125b	BE622841	SENP2	sentrin-specific protease	M + P
miR-125b	NM_003011	SET	SET translocation (myeloid	M + T	DNA replication\|endoplasmic
			leukemia-associated)		reticulum\|histone binding\|negative
					regulation of histone
					acetylation\|nucleocytoplasmic
					transport\|nucleosome
					assembly\|nucleosome
					disassembly\|nucleus\|perinuclear
					region\|protein phosphatase inhibitor
					activity\|protein phosphatase type 2A
					regulator activity
miR-125b	NM_006275	SFRS6	splicing factor,	P + T	RNA binding\|mRNA splice site
			arginine/serine-rich 6		selection\|nuclear mRNA splicing, via
					spliceosome\|nucleotide
					binding\|nucleus
miR-125b	AF015043	SH3BP4	SH3-domain binding protein 4	P + T	cell cycle\|endocytosis\|nucleus\|signal
					transducer activity
miR-125b	NM_016538	SIRT7	sirtuin silent mating type	P + T	DNA binding\|chromatin
			information regulation
2		silencing\|chromatin silencing
			homolog 7 (S. cerevisiae)		complex\|hydrolase
					activity\|regulation of transcription,
					DNA-dependent
miR-125b	NM_020309	SLC17A7	solute carrier family 17	P + T	integral to membrane\|phosphate
			(sodium-dependent		transport\|sodium-dependent
			inorganic phosphate		phosphate transporter
			cotransporter), member 7		activity\|transport\|transporter activity
miR-125b	NM_013272	SLC21A11	solute carrier family 21	P + T	integral to membrane\|ion
			(organic anion transporter),		transport\|membrane\|transporter
			member
11		activity
miR-125b	AK000722	SLC27A4	solute carrier family 27	P + T	catalytic activity\|fatty acid
			(fatty acid transporter),		transport\|fatty acid transporter
			member 4		activity\|ligase activity\|lipid
					metabolism\|lipid
					transport\|metabolism
miR-125b	NM_003759	SLC4A4	solute carrier family 4,	P + T	anion transport\|inorganic anion
			sodium bicarbonate		exchanger activity\|integral to
			cotransporter, member 4		membrane\|integral to plasma
					membrane\|membrane\|sodium:bicarbonate
					symporter activity\|transport
miR-125b	NM_003045	SLC7A1	solute carrier family 7	P + T	amino acid metabolism\|amino acid
			(cationic amino acid		permease activity\|amino acid
			transporter, y+ system),		transport\|basic amino acid
			member
1		transporter activity\|integral to plasma
					membrane\|membrane\|receptor
					activity\|transport
miR-125b	NM_003983	SLC7A6	solute carrier family 7	P + T	amino acid metabolism\|amino acid
			(cationic amino acid		transport\|amino acid-polyamine
			transporter, y+ system),		transporter activity\|integral to plasma
			member
6		membrane\|plasma membrane\|protein
					complex assembly\|transport
miR-125b	AF113019	SMARCD2	SWI/SNF related, matrix	M + P + T	chromatin
			associated, actin dependent		remodeling\|nucleoplasm\|regulation
			regulator of chromatin,		of transcription from RNA
			subfamily d, member 2		polymerase II
					promoter\|transcription\|transcription
					coactivator activity
miR-125b	NM_005985	SNAI1	snail homolog 1	P + T	DNA binding\|cartilage
			(Drosophila)		condensation\|development\|neuro-
					genesis\|nucleus\|zinc ion binding
miR-125b	AB037750	SORCS2	VPS10 domain receptor	P + T	integral to membrane\|intracellular
			protein		protein
					transport\|membrane\|membrane\|neuropeptide
					receptor
					activity\|neuropeptide signaling
					pathway\|protein binding\|protein
					transporter activity\|sugar binding
miR-125b	BE742268	SORT1	sortilin 1	P + T	endocytosis\|endosome\|integral to
					membrane\|integral to
					membrane\|intracellular protein
					transport\|membrane\|neurotensin
					receptor activity, G-protein
					coupled\|protein transporter
					activity\|receptor activity
miR-125b	AI360875	SOX11	SRY (sex determining	M + T	DNA
			region Y)-box 11		binding\|neurogenesis\|nucleus\|regulation
					of transcription, DNA-
					dependent\|transcription
miR-125b	AU121035	SP1	Sp1 transcription factor	P + T	DNA binding\|RNA polymerase II
					transcription factor
					activity\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcriptional
					activator activity\|zinc ion binding
miR-125b	NM_003131	SRF	serum response factor (c-fos	M + T	RNA polymerase II transcription
			serum response element		factor activity\|nucleus\|regulation of
			binding transcription factor)		transcription from RNA polymerase
					II promoter\|signal
					transduction\|transcription\|transcription
					factor activity
miR-125b	NM_005637	SS18	synovial sarcoma	P + T	nucleus
			translocation, chromosome 18
miR-125b	AF343880	SSX2	synovial sarcoma, X	P + T	nucleus
			breakpoint
2
miR-125b	NM_014682	ST18	suppression of	P + T	nucleus\|regulation of transcription,
			tumorigenicity 18 (breast		DNA-dependent\|transcription factor
			carcinoma) (zinc finger		activity
			protein)
miR-125b	AA128023	STARD13	START domain containing 13	P + T
miR-125b	BC000627	STAT3	signal transducer and	P + T	JAK-STAT cascade\|acute-phase
			activator of transcription 3		response\|calcium ion binding\|cell
			(acute-phase response		motility\|cytoplasm\|hematopoietin\|interferon-
			factor)		class (D200-domain)
					cytokine receptor signal transducer
					activity\|intracellular signaling
					cascade\|negative regulation of
					transcription from RNA polymerase II
					promoter\|neurogenesis\|nucleus\|nu-
					cleus\|regulation of transcription, DNA-
					dependent\|signal transducer
					activity\|transcription\|transcription
					factor activity\|transcription factor
					activity
miR-125b	NM_003155	STC1	stanniocalcin 1	P + T	calcium ion homeostasis\|cell surface
					receptor linked signal
					transduction\|cell-cell
					signaling\|extracellular
					region\|hormone activity\|response to
					nutrients
miR-125b	NM_003173	SUV39H1	suppressor of variegation 3-	P + T	DNA replication and chromosome
			9 homolog 1 (Drosophila)		cycle\|S-adenosylmethionine-
					dependent methyltransferase
					activity\|chromatin\|chromatin
					assembly or disassembly\|chromatin
					binding\|chromatin
					modification\|condensed nuclear
					chromosome\|histone lysine N-
					methyltransferase activity (H3-K9
					specific)\|histone-lysine N-
					methyltransferase
					activity\|methyltransferase
					activity\|nucleus\|nucleus\|protein
					binding\|transferase activity\|zinc ion
					binding
miR-125b	AW139618	SYN2	synapsin II	P + T	neurotransmitter
					secretion\|synapse\|synaptic
					transmission\|synaptic vesicle
miR-125b	R60550	TAF5L	TAF5-like RNA polymerase	M + P + T	nucleus\|regulation of transcription,
			II, p300/CBP-associated		DNA-dependent\|transcription factor
			factor (PCAF)-associated		activity\|transcription from RNA
			factor, 65 kDa		polymerase II promoter
miR-125b	AF220509	TAF9L	TAF9-like RNA polymerase	P + T	DNA binding\|nucleus\|regulation of
			II, TATA box binding		transcription, DNA-
			protein (TBP)-associated		dependent\|transcription factor TFIID
			factor, 31 kDa		complex\|transcription initiation
miR-125b	NM_000116	TAZ	tafazzin (cardiomyopathy,	M + P + T	acyltransferase activity\|heart
			dilated 3A (X-linked);		development\|integral to
			endocardial fibroelastosis 2;		membrane\|metabolism\|muscle
			Barth syndrome)		contraction\|muscle development
miR-125b	NM_018488	TBX4	T-box 4	P + T	development\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					factor activity
miR-125b	NM_012249	TC10	ras-like protein TC10	M + T	GTP binding\|GTPase activity\|plasma
					membrane\|small GTPase mediated
					signal transduction
miR-125b	BG387172	TEAD2	TEA domain family	P + T	nucleus\|nucleus\|regulation of
			member 2		transcription, DNA-
					dependent\|regulation of transcription,
					DNA-
					dependent\|transcription\|transcription
					factor activity\|transcription factor
					activity
miR-125b	U06935	TEF	thyrotrophic embryonic	P + T	RNA polymerase II transcription
			factor		factor activity\|nucleus\|regulation of
					transcription from RNA polymerase
					II promoter\|rhythmic
					process\|transcription\|transcription
					factor activity
miR-125b	NM_006464	TGOLN2	trans-golgi network protein 2	P + T	Golgi trans face\|integral to
					membrane\|transport vesicle
miR-125b	BE219311	TIMM22	translocase of inner	P + T	integral to membrane\|mitochondrial
			mitochondrial membrane 22		inner
			homolog (yeast)		membrane\|mitochondrion\|protein
					transport\|protein transporter activity
miR-125b	NM_003326	TNFSF4	tumor necrosis factor	P + T	cell-cell signaling\|immune
			(ligand) superfamily,		response\|integral to plasma
			member 4 (tax-		membrane\|membrane\|positive
			transcriptionally activated		regulation of cell proliferation\|signal
			glycoprotein
1, 34 kDa)		transduction\|tumor necrosis factor
					receptor binding
miR-125b	AA873275	TOR2A	torsin family	2, member A	P + T	ATP binding\|GTP cyclohydrolase I
					activity\|biosynthesis\|chaperone
					cofactor dependent protein
					folding\|endoplasmic
					reticulum\|nucleoside-triphosphatase
					activity\|nucleotide binding
miR-125b	AW341649	TP53INP1	tumor protein p53 inducible	M + P + T	apoptosis\|nucleus
			nuclear protein 1
miR-125b	NM_014112	TRPS1	trichorhinophalangeal	P + T	NLS-bearing substrate-nucleus
			syndrome I		import\|nucleus\|regulation of
					transcription, DNA-
					dependent\|skeletal
					development\|transcription\|transcription
					factor activity\|transcription from
					RNA polymerase II promoter\|zinc
					ion binding
miR-125b	NM_001070	TUBG1	tubulin, gamma 1	P + T	GTP binding\|GTPase
					activity\|centrosome\|condensed
					nuclear chromosome\|gamma-tubulin
					complex\|meiotic spindle
					organization and
					biogenesis\|microtubule\|microtubule
					nucleation\|microtubule-based
					movement\|mitotic spindle
					organization and biogenesis\|polar
					microtubule\|protein binding\|protein
					polymerization\|spindle pole
					body\|structural constituent of
					cytoskeleton
miR-125b	NM_003330	TXNRD1	thioredoxin reductase 1	P + T	FAD binding\|cell redox
					homeostasis\|cytoplasm\|disulfide
					oxidoreductase activity\|electron
					transport\|electron transporter
					activity\|oxidoreductase activity,
					acting on NADH or NADPH,
					disulfide as acceptor\|signal
					transduction\|thioredoxin-disulfide
					reductase activity
miR-125b	BC004862	UBE2R2	ubiquitin-conjugating	P + T	ligase activity\|ubiquitin conjugating
			enzyme E2R 2		enzyme activity\|ubiquitin
					cycle\|ubiquitin-protein ligase activity
miR-125b	NM_003728	UNC5C	unc-5 homolog B	P + T	apoptosis\|axon guidance\|brain
			(C. elegans)		development\|development\|integral to
					membrane\|netrin receptor
					activity\|protein binding\|receptor
					activity\|signal transduction
miR-125b	NM_003369	UVRAG	UV radiation resistance	P + T	DNA repair\|cytoplasm
			associated gene
miR-125b	AF195514	VPS4B	vacuolar protein sorting 4B	M + P + T	ATP binding\|ATPase activity,
			(yeast)		coupled\|membrane\|membrane
					fusion\|nucleoside-triphosphatase
					activity\|nucleotide
					binding\|peroxisome organization and
					biogenesis\|protein binding\|regulation
					of transcription, DNA-dependent
miR-125b	R51061	VTS58635	mitogen-activated protein	P + T	GTP binding\|small GTPase mediated
			kinase kinase kinase kinase 1		signal transduction
miR-125b	NM_004184	WARS	tryptophanyl-tRNA	M + T	ATP binding\|cytoplasm\|ligase
			synthetase		activity\|negative regulation of cell
					proliferation\|protein
					biosynthesis\|soluble
					fraction\|tryptophan-tRNA ligase
					activity\|tryptophanyl-tRNA
					aminoacylation\|tryptophanyl-tRNA
					aminoacylation
miR-125b	NM_005433	YES1	v-yes-1 Yamaguchi sarcoma	P + T	ATP binding\|intracellular signaling
			viral oncogene homolog 1		cascade\|protein amino acid
					phosphorylation\|protein-tyrosine
					kinase activity\|transferase activity
miR-125b	NM_017740	ZDHHC7	zinc finger, DHHC domain	P + T	integral to membrane\|metal ion
			containing 7		binding
miR-125b	BF525395	ZFP385	likely ortholog of mouse	M + P + T	DNA binding\|nucleic acid
			zinc finger protein 385		binding\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|zinc ion
					binding
miR-125b	NM_007345	ZNF236	zinc finger protein 236	P + T	nucleus\|regulation of transcription,
					DNA-
					dependent\|transcription\|transcription
					factor activity\|zinc ion binding
miR-125b	NM_012482	ZNF281	zinc finger protein 281	M + P + T	DNA binding\|DNA-directed RNA
					polymerase II, core
					complex\|negative regulation of
					transcription from RNA polymerase
					II promoter\|nucleus\|regulation of
					transcription, DNA-
					dependent\|specific RNA polymerase
					II transcription factor
					activity\|transcription\|zinc ion binding
miR-125b	NM_003427	ZNF76	zinc finger protein 76	P + T	DNA binding\|nucleus\|regulation of
			(expressed in testis)		transcription from RNA polymerase
					II promoter\|regulation of
					transcription from RNA polymerase
					III promoter\|transcription\|zinc ion
					binding
miR-125b	NM_022465	ZNFN1A4	zinc finger protein,	M + P + T	nucleic acid
			subfamily 1A, 4 (Eos)		binding\|nucleus\|transcription factor
					activity\|transcriptional repressor
					activity\|zinc ion binding
miR-145	NM_005502	ABCA1	ATP-binding cassette, sub-	P + T	ATP binding\|ATP binding\|ATPase
			family A (ABC1), member 1		activity\|anion transporter
					activity\|cholesterol
					metabolism\|integral to plasma
					membrane\|lipid
					metabolism\|membrane
					fraction\|nucleotide binding\|steroid
					metabolism\|sterol transporter
					activity\|transport\|transport
miR-145	AL527773	ABR	active BCR-related gene	M + P + T	GTPase activator activity\|guanyl-
					nucleotide exchange factor
					activity\|small GTPase mediated
					signal transduction
miR-145	NM_001616	ACVR2	activin A receptor, type II	M + P + T	ATP binding\|integral to plasma
					membrane\|membrane\|protein amino
					acid phosphorylation\|receptor
					activity\|transferase
					activity\|transforming growth factor
					beta receptor activity\|transmembrane
					receptor protein serine/threonine
					kinase signaling pathway
miR-145	NM_003183	ADAM17	a disintegrin and	P + T	cell-cell signaling\|integral to plasma
			metalloproteinase domain		membrane\|metalloendopeptidase
			17 (tumor necrosis factor,		activity\|proteolysis and
			alpha, converting enzyme)		peptidolysis\|zinc ion binding
miR-145	NM_019903	ADD3	adducin 3 (gamma)	M + P + T	calmodulin
					binding\|cytoskeleton\|membrane\|structural
					constituent of cytoskeleton
miR-145	AB003476	AKAP12	A kinase (PRKA) anchor	P + T	G-protein coupled receptor protein
			protein (gravin) 12		signaling pathway\|cytoplasm\|protein
					binding\|protein kinase A
					binding\|protein targeting\|signal
					transduction
miR-145	NM_016201	AMOTL2	angiomotin like 2	M + P + T
miR-145	NM_001128	AP1G1	adaptor-related protein	M + P + T	Golgi apparatus\|binding\|clathrin coat
			complex
1, gamma 1 subunit		of trans-Golgi network vesicle\|coated
					pit\|endocytosis\|intracellular protein
					transport\|intracellular protein
					transport\|membrane coat adaptor
					complex\|protein complex
					assembly\|transporter activity
miR-145	NM_001284	AP3S1	adaptor-related protein	M + P + T	Golgi apparatus\|clathrin vesicle
			complex
3, sigma 1 subunit		coat\|insulin receptor signaling
					pathway\|intracellular protein
					transport\|membrane coat adaptor
					complex\|transport\|transport
					vesicle\|transporter activity
miR-145	NM_006380	APPBP2	amyloid beta precursor	M + P + T	binding\|cytoplasm\|intracellular
			protein (cytoplasmic tail)		protein
			binding protein
2		transport\|membrane\|microtubule
					associated complex\|microtubule
					motor activity\|nucleus
miR-145	AB037845	ARHGAP10	Rho-GTPase activating	M + T	protein binding
			protein 10
miR-145	AL516350	ARPC5	actin related protein 2/3	P + T	Arp2/3 protein complex\|actin
			complex, subunit 5, 16 kDa		cytoskeleton organization and
					biogenesis\|cell
					motility\|cytoplasm\|cytoskeleton\|regulation
					of actin filament
					polymerization\|structural constituent
					of cytoskeleton
miR-145	U72937	ATRX	alpha thalassemia/mental	M + T	ATP binding\|DNA binding\|DNA
			retardation syndrome X-		helicase activity\|DNA
			linked (RAD54 homolog,		methylation\|DNA
			S. cerevisiae)		recombination\|DNA
					repair\|chromosome organization and
					biogenesis (sensu
					Eukaryota)\|helicase
					activity\|hydrolase activity\|nuclear
					heterochromatin\|nucleus\|perception
					of sound\|regulation of transcription,
					DNA-dependent\|transcription factor
					activity
miR-145	NM_021813	BACH2	BTB and CNC homology 1,	P + T	DNA binding\|nucleus\|protein
			basic leucine zipper		binding\|regulation of transcription,
			transcription factor 2		DNA-dependent\|transcription
miR-145	NM_013449	BAZ2A	bromodomain adjacent to	P + T	DNA binding\|chromatin
			zinc finger domain, 2A		remodeling\|nucleolus organizer
					complex\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					regulator activity
miR-145	NM_007005	BCE-1	BCE-1 protein	M + P	frizzled signaling
					pathway\|molecular_function
					unknown\|nucleus\|nucleus\|regulation
					of transcription\|regulation of
					transcription, DNA-dependent
miR-145	NM_003458	BSN	bassoon (presynaptic	P + T	cytoskeleton\|metal ion
			cytomatrix protein)		binding\|nucleus\|structural constituent
					of cytoskeleton\|synapse\|synaptic
					transmission\|synaptosome
miR-145	NM_013279	C11orf9	chromosome 11 open	M + P + T
			reading frame
9
miR-145	NM_024643	C14orf140	hypothetical protein	P + T
			FLJ23093
miR-145	NM_018270	C20orf20	chromosome 20 open	P + T	chromatin
			reading frame
20		modification\|nucleus\|regulation of
					cell growth\|regulation of
					transcription, DNA-
					dependent\|transcription
miR-145	NM_004276	CABP1	calcium binding protein 1	P + T	calcium ion binding\|calcium ion
			(calbrain)		binding\|enzyme inhibitor activity
miR-145	NM_001755	CBFB	core-binding factor, beta	M + P + T	RNA polymerase II transcription
			subunit		factor activity\|nucleus\|transcription
					coactivator activity\|transcription
					factor activity\|transcription from
					RNA polymerase II promoter
miR-145	NM_001759	CCND2	cyclin D2	P + T	cytokinesis\|nucleus\|regulation of cell
					cycle
miR-145	NM_020307	CCNL1	cyclin L ania-6a	M + P + T	cell cycle\|regulation of cell cycle
miR-145	AL118798	CD47	CD47 antigen (Rh-related	P + T	cell-matrix adhesion\|integral to
			antigen, integrin-associated		plasma membrane\|integrin-mediated
			signal transducer)		signaling pathway\|plasma
					membrane\|protein binding
miR-145	BF576053	CFL2	cofilin 2 (muscle)	M + P + T	actin binding\|cytoskeleton\|nucleus
miR-145	AA835485	CKLiK	CamKI-like protein kinase	P + T	ATP binding\|calcium- and
					calmodulin-dependent protein kinase
					activity\|calmodulin
					binding\|nucleus\|protein amino acid
					phosphorylation\|protein
					serine/threonine kinase
					activity\|transferase activity
miR-145	NM_004921	CLCA3	chloride channel, calcium	P + T	extracellular
			activated, family member 3		space\|transport\|transporter activity
miR-145	NM_001326	CSTF3	cleavage stimulation factor,	M + P + T	RNA binding\|binding\|mRNA
			3′ pre-RNA, subunit 3,		cleavage\|mRNA
			77 kDa		polyadenylylation\|nucleus
miR-145	NM_020248	CTNNBIP1	catenin, beta interacting	P + T	Wnt receptor signaling pathway\|beta-
			protein 1		catenin binding\|cell
					proliferation\|development\|nucleus\|regulation
					of transcription, DNA-
					dependent\|signal transduction
miR-145	AW772082	DACH	dachshund homolog	P + T	DNA binding\|development\|eye
			(Drosophila)		morphogenesis (sensu
					Endopterygota)\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription
miR-145	NM_004393	DAG1	dystroglycan 1 (dystrophin-	M + P + T	actin cytoskeleton\|calcium ion
			associated glycoprotein 1)		binding\|extracellular matrix (sensu
					Metazoa)\|integral to plasma
					membrane\|laminin receptor
					activity\|membrane fraction\|muscle
					contraction\|plasma
					membrane\|protein binding\|protein
					complex assembly
miR-145	NM_003887	DDEF2	development and	P + T	GTPase activator activity\|Golgi
			differentiation enhancing		apparatus\|regulation of GTPase
			factor
2		activity
miR-145	AL080239	DKFZp547M2010	hypothetical protein	M + P + T
			DKFZp547M2010
miR-145	AL137517	DKFZp564O1278	hypothetical protein	P + T	integral to membrane
			DKFZp564O1278
miR-145	NM_001386	DPYSL2	dihydropyrimidinase-like 2	P + T	dihydropyrimidinase
					activity\|hydrolase
					activity\|neurogenesis\|nucleobase,
					nucleoside, nucleotide and nucleic
					acid metabolism\|signal transduction
miR-145	BC003143	DUSP6	dual specificity phosphatase 6	P + T	MAP kinase phosphatase
					activity\|cytoplasm\|hydrolase
					activity\|inactivation of
					MAPK\|protein amino acid
					dephosphorylation\|protein
					serine/threonine phosphatase
					activity\|protein tyrosine phosphatase
					activity\|regulation of cell
					cycle\|soluble fraction
miR-145	D86550	DYRK1A	dual-specificity tyrosine-	P + T	ATP
			(Y)-phosphorylation		binding\|neurogenesis\|nucleus\|protein
			regulated kinase 1A		amino acid phosphorylation\|protein
					serine/threonine kinase
					activity\|protein-tyrosine kinase
					activity\|transferase activity
miR-145	NM_001967	EIF4A2	eukaryotic translation	M + P + T	ATP binding\|ATP-dependent
			initiation factor 4A, isoform 2		helicase activity\|DNA binding\|RNA
					binding\|eukaryotic translation
					initiation factor 4F
					complex\|hydrolase activity\|protein
					biosynthesis\|regulation of
					translational initiation\|translation
					initiation factor activity
miR-145	NM_001417	EIF4B	eukaryotic translation	M + T	RNA binding\|eukaryotic translation
			initiation factor 4B		initiation factor 4F complex\|nucleic
					acid binding\|nucleotide
					binding\|protein
					biosynthesis\|regulation of
					translational initiation\|translation
					initiation factor activity\|translation
					initiation factor activity
miR-145	BC005057	EIF4EBP2	eukaryotic translation	P + T	eukaryotic initiation factor 4E
			initiation factor 4E binding		binding\|negative regulation of
			protein 2		protein biosynthesis\|negative
					regulation of translational
					initiation\|regulation of translation
miR-145	NM_020909	EPB41L5	erythrocyte membrane	P + T	binding\|cytoplasm\|cytoskeletal
			protein band 4.1 like 5		protein
					binding\|cytoskeleton\|membrane
miR-145	NM_005797	EVA1	epithelial V-like antigen 1	P + T	cell
					adhesion\|cytoskeleton\|homophilic
					cell adhesion\|integral to
					membrane\|membrane\|morpho-
					genesis\|protein binding
miR-145	NM_022977	FACL4	fatty-acid-Coenzyme A	M + P + T	fatty acid metabolism\|integral to
			ligase, long-chain 4		membrane\|learning and/or
					memory\|ligase activity\|lipid
					metabolism\|long-chain-fatty-acid-
					CoA ligase activity\|magnesium ion
					binding\|metabolism
miR-145	AL042120	FHOD2	formin homology	2 domain	M + P	Rho GTPase binding\|actin
			containing 2		binding\|actin cytoskeleton
					organization and biogenesis\|cell
					organization and
					biogenesis\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription factor
					activity\|translation initiation factor
					activity\|translational initiation
miR-145	NM_002013	FKBP3	FK506 binding protein 3,	P + T	FK506 binding\|isomerase
			25 kDa		activity\|nucleus\|peptidyl-prolyl cis-
					trans isomerase activity\|protein
					folding\|receptor activity
miR-145	NM_002017	FLI1	Friend leukemia virus	M + P + T	hemostasis\|nucleus\|organogenesis\|regulation
			integration
1		of transcription, DNA-
					dependent\|transcription\|transcription
					factor activity
miR-145	NM_023071	FLJ13117	hypothetical protein	P + T
			FLJ13117
miR-145	AL561281	FLJ20373	hypothetical protein	M + P + T	ATP binding\|cellular_component
			FLJ20373		unknown\|protein amino acid
					phosphorylation\|protein kinase
					cascade\|protein serine/threonine
					kinase activity\|response to
					stress\|signal transduction\|small
					GTPase regulator activity\|transferase
					activity
miR-145	AK025444	FLJ21791	hypothetical protein	M + T
			FLJ21791
miR-145	NM_024713	FLJ22557	hypothetical protein	P + T
			FLJ22557
miR-145	AA872588	FLJ36155	likely ortholog of mouse	P + T	DNA binding\|negative regulation of
			Gli-similar 1 Kruppel-like		transcription from RNA polymerase
			zinc finger (Glis1)		II promoter\|nucleus\|positive
					regulation of transcription from RNA
					polymerase II promoter\|regulation of
					transcription, DNA-
					dependent\|specific RNA polymerase
					II transcription factor
					activity\|transcription\|zinc ion binding
miR-145	AI434509	FLJ38499	Unnamed protein product	P + T	nucleic acid binding
			[Homo sapiens], mRNA
			sequence
miR-145	M62994	FLNB	filamin B, beta (actin	P + T	actin binding\|actin binding\|actin
			binding protein 278)		cytoskeleton\|actin cytoskeleton
					organization and biogenesis\|cell
					differentiation\|cytoskeletal
					anchoring\|integral to plasma
					membrane\|myogenesis\|signal
					transduction
miR-145	NM_002025	FMR2	fragile X mental retardation 2	M + T	brain development\|learning and/or
					memory
miR-145	N29672	FOS	v-fos FBJ murine	M + T	proto-oncogene
			osteosarcoma viral
			oncogene homolog
miR-145	NM_002015	FOXO1A	forkhead box O1A	M + P + T	anti-apoptosis\|nucleus\|regulation of
			(rhabdomyosarcoma)		transcription from RNA polymerase II
					promoter\|transcription\|transcription
					factor activity
miR-145	NM_003507	FZD7	frizzled homolog 7	M + P + T	G-protein coupled receptor
			(Drosophila)		activity\|G-protein coupled receptor
					protein signaling pathway\|Wnt
					receptor
					activity\|development\|frizzled
					signaling pathway\|integral to
					membrane\|plasma membrane
miR-145	AL049709	GGTL3	gamma-glutamyltransferase-	M + P + T
			like 3
miR-145	NM_022735	GOCAP1	golgi complex associated	M + P + T	Golgi apparatus\|acyl- CoA
			protein

1, 60 kDa		binding\|catalytic activity\|intracellular
					protein
					transport\|membrane\|mitochondrion\|protein
					carrier activity\|steroid
					biosynthesis
miR-145	NM_020806	GPHN	gephyrin	P + T	Mo-molybdopterin cofactor
					biosynthesis\|catalytic
					activity\|cytoskeleton
miR-145	NM_015071	GRAF	GTPase regulator associated	P + T	Rho GTPase activator activity\|actin
			with focal adhesion kinase		cytoskeleton organization and
			pp125(FAK)		biogenesis\|cellular_component
					unknown\|neurogenesis
miR-145	NM_017913	HARC	Hsp90-associating relative	P + T	cytokinesis\|regulation of cell cycle
			of Cdc37
miR-145	BC006237	HECTD1	HECT domain containing 1	M + T	intracellular\|ligase activity\|receptor
					activity\|ubiquitin cycle\|ubiquitin-
					protein ligase activity
miR-145	U64317	HEF1	enhancer of filamentation 1	P + T	actin filament bundle formation\|cell
			(cas-like docking; Crk-		adhesion\|cytokinesis\|cytoplasm\|cyto-
			associated substrate related)		skeleton\|cytoskeleton organization
					and biogenesis\|integrin-mediated
					signaling
					pathway\|mitosis\|nucleus\|protein
					binding\|regulation of cell
					cycle\|regulation of cell growth\|signal
					transduction\|spindle
miR-145	NM_016258	HGRG8	high-glucose-regulated	P + T
			protein 8
miR-145	AL162003	HIC2	hypermethylated in cancer 2	P + T	DNA binding\|negative regulation of
					transcription, DNA-
					dependent\|nucleus\|protein C-
					terminus binding\|transcription\|zinc
					ion binding
miR-145	NM_014212	HOXC11	homeo box C11	M + P + T	RNA polymerase II transcription
					factor
					activity\|development\|endoderm
					development\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription factor
					activity
miR-145	NM_002193	INHBB	inhibin, beta B (activin AB	M + P + T	cell differentiation\|cytokine
			beta polypeptide)		activity\|defense
					response\|extracellular
					region\|growth\|growth factor
					activity hormone activity\|host cell
					surface receptor binding\|negative
					regulation of follicle-stimulating
					hormone secretion\|negative
					regulation of hepatocyte growth
					factor biosynthesis\|ovarian follicle
					development\|positive regulation of
					follicle-stimulating hormone
					secretion\|protein binding\|protein
					homodimerization activity\|response
					to external stimulus
miR-145	NM_005544	IRS1	insulin receptor substrate 1	M + P + T	cytoplasm\|insulin receptor
					binding\|protein binding\|signal
					transducer activity\|signal
					transduction\|transmembrane receptor
					protein tyrosine kinase docking
					protein activity
miR-145	NM_006459	KEO4	similar to Caenorhabditis	P + T	catalytic activity
			elegans protein C42C1.9
miR-145	NM_014686	KIAA0355	KIAA0355 gene product	P + T
miR-145	NM_015176	KIAA0483	KIAA0483 protein	P + T	ubiquitin cycle
miR-145	NM_014871	KIAA0710	KIAA0710 gene product	M + P + T	cysteine-type endopeptidase
					activity\|exonuclease
					activity\|nucleus\|ubiquitin
					cycle\|ubiquitin thiolesterase
					activity\|ubiquitin-dependent protein
					catabolism
miR-145	AA772278	KIAA1673	KIAA1673	M + P + T
miR-145	AB051495	KIAA1708	KIAA1708 protein	P + T	ATP binding\|microtubule associated
					complex\|microtubule motor
					activity\|microtubule-based
					movement
miR-145	AI814587	KIAA1715	KIAA1715 protein	M + T
miR-145	AI187364	KIAA1894	KIAA1894 protein	P + T	integral to membrane
miR-145	AF155117	KIF21A	kinesin family member 21A	P + T	ATP binding\|microtubule associated
					complex\|microtubule motor
					activity\|microtubule-based
					movement
miR-145	NM_004235	KLF4	Kruppel-like factor 4 (gut)	M + T	mesodermal cell fate
					determination\|negative regulation of
					cell proliferation\|negative regulation
					of transcription, DNA-
					dependent\|negative regulation of
					transcription, DNA-
					dependent\|nucleic acid
					binding\|nucleus\|transcription\|transcription
					factor activity\|transcription
					factor activity\|transcriptional
					activator activity\|transcriptional
					activator activity\|transcriptional
					repressor activity\|transcriptional
					repressor activity\|zinc ion
					binding\|zinc ion binding
miR-145	T68150	LL5beta	hypothetical protein	M + T
			FLJ21791
miR-145	AI797833	LOC285148	a disintegrin and	P + T	catalytic activity
			metalloproteinase domain
			17 (tumor necrosis factor,
			alpha, converting enzyme)
miR-145	NM_025146	MAK3P	likely ortholog of mouse	P + T	N-acetyltransferase activity
			Mak3p homolog
			(S. cerevisiae)
miR-145	BF971923	MAP3K3	mitogen-activated protein	M + P	ATP binding\|MAP kinase kinase
			kinase kinase kinase 3		kinase activity\|MAPKKK
					cascade\|magnesium ion
					binding\|positive regulation of I-
					kappaB kinase/NF-kappaB
					cascade\|protein amino acid
					phosphorylation\|protein kinase
					activity\|protein serine/threonine
					kinase activity\|signal transducer
					activity\|transferase activity
miR-145	NM_004834	MAP4K4	mitogen-activated protein	M + P + T	ATP binding\|cellular_component
			kinase kinase kinase kinase 4		unknown\|protein amino acid
					phosphorylation\|protein kinase
					cascade\|protein serine/threonine
					kinase activity\|response to
					stress\|signal transduction\|small
					GTPase regulator activity\|transferase
					activity
miR-145	BF382281	MGC10120	Homo sapiens cDNA	P + T
			FLJ30135 fis, clone
			BRACE2000061, mRNA
			sequence
miR-145	BG231756	MGC10986	hypothetical protein	M + P	ATP binding\|MAP kinase kinase
			MGC10986		kinase activity\|MAPKKK
					cascade\|magnesium ion
					binding\|positive regulation of I-
					kappaB kinase/NF-kappaB
					cascade\|protein amino acid
					phosphorylation\|protein kinase
					activity\|protein serine/threonine
					kinase activity\|signal transducer
					activity\|transferase activity
miR-145	BC004869	MGC2817	hypothetical protein	P + T	outer membrane\|protein transport
			MGC2817
miR-145	BC002712	MYCN	v-myc myelocytomatosis	M + T	chromatin\|nucleus\|protein
			viral related oncogene,		binding\|regulation of transcription
			neuroblastoma derived		from RNA polymerase II
			(avian)		promoter\|transcription factor activity
miR-145	AB007899	NEDD4L	neural precursor cell	P + T	excretion\|intracellular\|intracellular\|ligase
			expressed, developmentally		activity\|positive regulation of
			down-regulated 4-like		endocytosis\|protein binding\|protein
					ubiquitination\|regulation of protein
					catabolism\|response to metal
					ion\|sodium channel regulator
					activity\|sodium ion
					homeostasis\|sodium ion
					transport\|ubiquitin cycle\|ubiquitin-
					protein ligase activity\|ubiquitin-
					protein ligase activity\|water
					homeostasis
miR-145	NM_005863	NET1	neuroepithelial cell	P + T	guanyl-nucleotide exchange factor
			transforming gene
1		activity\|nucleus\|regulation of cell
					growth\|signal transduction
miR-145	NM_003204	NFE2L1	nuclear factor (erythroid-	P + T	DNA binding\|heme
			derived 2)-like 1		biosynthesis\|inflammatory
					response\|morphogenesis\|nucleus\|nu-
					cleus\|regulation of transcription, DNA-
					dependent\|transcription\|transcription
					cofactor activity\|transcription factor
					activity\|transcription from RNA
					polymerase II promoter
miR-145	NM_006469	NS1-BP	NS1-binding protein	M + P + T	RNA splicing\|protein
					binding\|response to
					virus\|spliceosome
					complex\|transcription factor
					complex\|transcription from RNA
					polymerase III promoter
miR-145	NM_019094	NUDT4	nudix (nucleoside	P + T	calcium-mediated signaling/cyclic
			diphosphate linked moiety		nucleotide metabolism cyclic-
			X)-type motif 4		nucleotide-mediated
					signaling\|diphosphoinositol-
					polyphosphate diphosphatase
					activity\|hydrolase
					activity\|intracellular\|intracellular
					signaling cascade\|intracellular
					transport\|magnesium ion
					binding\|regulation of RNA-nucleus
					export
miR-145	AW149417	OAZ	OLF-1/EBF associated zinc	P + T	nucleic acid binding\|nucleus\|zinc ion
			finger gene		binding
miR-145	NM_024586	OSBPL9	oxysterol binding protein-	M + P	lipid transport\|steroid metabolism
			like 9
miR-145	AB040812	PAK7	p21(CDKN1A)-activated	M + T	ATP binding protein amino acid
			kinase 7		phosphorylation\|protein
					serine/threonine kinase
					activity\|transferase activity
miR-145	NM_014456	PDCD4	programmed cell death 4	M + P + T	apoptosis
			(neoplastic transformation
			inhibitor)
miR-145	NM_002657	PLAGL2	pleiomorphic adenoma	M + P + T	nucleus\|regulation of transcription,
			gene-like 2		DNA-
					dependent\|transcription\|transcription
					factor activity\|zinc ion binding
miR-145	AK023546	PLCL2	phospholipase C-like 2	P + T	calcium ion binding\|intracellular
					signaling cascade\|lipid
					metabolism\|phosphoinositide
					phospholipase C activity
miR-145	AI274352	PLN	phospholamban	P + T
miR-145	NM_000944	PPP3CA	protein phosphatase 3	P + T	calcineurin complex\|calcium ion
			(formerly 2B), catalytic		binding\|calmodulin
			subunit, alpha isoform		binding\|hydrolase activity\|protein
			(calcineurin A alpha)		amino acid
					dephosphorylation\|protein
					serine/threonine phosphatase activity
miR-145	BF247371	PRO1843	hypothetical protein	M + T
			PRO1843
miR-145	NM_000959	PTGFR	prostaglandin F receptor	P + T	G-protein coupled receptor protein
			(FP)		signaling pathway\|G-protein coupled
					receptor protein signaling
					pathway\|integral to
					membrane\|integral to plasma
					membrane\|parturition\|prostaglandin
					F receptor activity\|prostaglandin F
					receptor activity\|receptor
					activity\|rhodopsin-like receptor
					activity\|signal
					transduction\|thromboxane receptor
					activity
miR-145	NM_002890	RASA1	RAS p21 protein activator	P + T	Ras GTPase activator
			(GTPase activating protein) 1		activity\|intracellular signaling
					cascade
miR-145	NM_006506	RASA2	RAS p21 protein activator 2	P + T	Ras GTPase activator
					activity\|intracellular signaling
					cascade
miR-145	NM_002912	REV3L	REV3-like, catalytic subunit	M + P + T	3′-5′ exonuclease activity\|DNA
			of DNA polymerase zeta		binding\|DNA repair\|DNA
			(yeast)		replication\|DNA-dependent DNA
					replication\|DNA-directed DNA
					polymerase activity\|nucleotide
					binding\|nucleus\|transferase
					activity\|zeta DNA polymerase
					activity\|zeta DNA polymerase
					complex
miR-145	NM_002924	RGS7	regulator of G-protein	P + T	heterotrimeric G-protein
			signalling 7		complex\|intracellular signaling
					cascade\|regulation of G-protein
					coupled receptor protein signaling
					pathway\|regulator of G-protein
					signaling activity\|signal transducer
					activity
miR-145	AL136924	RIN2	Ras and Rab interactor 2	P + T	GTPase activator activity\|Rab
					guanyl-nucleotide exchange factor
					activity\|cellular_component
					unknown\|endocytosis\|intracellular
					signaling cascade\|small GTPase
					mediated signal transduction\|small
					GTPase regulator activity
miR-145	BE463945	RTKN	rhotekin	P + T	intracellular\|protein binding\|signal
					transduction\|signal transduction
miR-145	AF225986	SCN3A	sodium channel, voltage-	P + T	cation channel activity\|cation
			gated, type III, alpha		transport\|integral to
			polypeptide		membrane\|membrane\|sodium ion
					transport\|voltage-gated sodium
					channel activity\|voltage-gated
					sodium channel complex
miR-145	NM_006080	SEMA3A	sema domain,	P + T	cell differentiation\|extracellular
			immunoglobulin domain		region\|neurogenesis
			(Ig), short basic domain,
			secreted, (semaphorin) 3A
miR-145	NM_020796	SEMA6A	sema domain,	P + T	apoptosis\|axon\|axon guidance\|cell
			transmembrane domain		differentiation\|cell surface receptor
			(TM), and cytoplasmic		linked signal
			domain, (semaphorin) 6A		transduction\|cytoskeleton
					organization and
					biogenesis\|development\|integral to
					membrane\|membrane\|neurogenesis\|protein
					binding\|receptor activity
miR-145	NM_004171	SLC1A2	solute carrier family 1 (glial	P + T	L-glutamate transport\|L-glutamate
			high affinity glutamate		transporter activity\|dicarboxylic acid
			transporter), member 2		transport\|integral to
					membrane\|membrane\|membrane
					fraction\|sodium:dicarboxylate
					symporter activity\|symporter
					activity\|synaptic
					transmission\|transport
miR-145	NM_003759	SLC4A4	solute carrier family 4,	P + T	anion transport\|inorganic anion
			sodium bicarbonate		exchanger activity\|integral to
			cotransporter, member 4		membrane\|integral to plasma
					membrane\|membrane\|sodium:bicarbonate
					symporter activity\|transport
miR-145	NM_030918	SNX27	hypothetical protein My014	M + P + T	intracellular signaling
					cascade\|protein binding\|protein
					transport
miR-145	AI360875	SOX11	SRY (sex determining	M + T	DNA
			region Y)-box 11		binding\|neurogenesis\|nucleus\|regulation
					of transcription, DNA-
					dependent\|transcription
miR-145	NM_000346	SOX9	SRY (sex determining	P + T	DNA binding\|cartilage
			region Y)-box 9		condensation\|nucleus\|regulation of
			(campomelic dysplasia,		transcription from RNA polymerase
			autosomal sex-reversal)		II promoter\|skeletal
					development\|specific RNA
					polymerase II transcription factor
					activity\|transcription
miR-145	AK023899	SRGAP1	SLIT-ROBO Rho GTPase	P + T	GTPase activator activity
			activating protein
1
miR-145	NM_003155	STC1	stanniocalcin 1	M + T	calcium ion homeostasis\|cell surface
					receptor linked signal
					transduction\|cell-cell
					signaling\|extracellular
					region\|hormone activity\|response to
					nutrients
miR-145	BE219311	TIMM22	translocase of inner	M + P + T	integral to membrane\|mitochondrial
			mitochondrial membrane 22		inner
			homolog (yeast)		membrane\|mitochondrion\|protein
					transport\|protein transporter activity
miR-145	AA705845	TLE4	transducin-like enhancer of	M + P	frizzled signaling
			split 4 (E(sp1) homolog,		pathway\|molecular_function
			Drosophila)		unknown\|nucleus\|nucleus\|regulation
					of transcription\|regulation of
					transcription, DNA-dependent
miR-145	BC005016	TRIM2	tripartite motif-containing 2	P + T	cytoplasm\|myosin binding\|protein
					ubiquitination\|ubiquitin ligase
					complex\|ubiquitin-protein ligase
					activity\|zinc ion binding
miR-145	NM_025076	UXS1	UDP-glucuronate	M + P + T	carbohydrate metabolism\|isomerase
			decarboxylase
1		activity\|nucleotide-sugar metabolism
miR-145	NM_005433	YES1	v-yes-1 Yamaguchi sarcoma	P + T	ATP binding\|intracellular signaling
			viral oncogene homolog 1		cascade\|protein amino acid
					phosphorylation\|protein-tyrosine
					kinase activity\|transferase activity
miR-145	BC003128	ZDHHC9	zinc finger, DHHC domain	P + T	integral to membrane\|metal ion
			containing 9		binding
miR-155	NM_019903	ADD3	adducin 3 (gamma)	P + T	calmodulin
					binding\|cytoskeleton\|membrane\|structural
					constituent of cytoskeleton
miR-155	NM_020661	AICDA	activation-induced cytidine	P + T	B-cell
			deaminase		differentiation\|cellular_component
					unknown\|cytidine deaminase
					activity\|hydrolase activity\|mRNA
					processing\|zinc ion binding
miR-155	NM_007202	AKAP10	A kinase (PRKA) anchor	P + T	kinase activity\|mitochondrion\|protein
			protein 10		binding\|protein localization\|signal
					transducer activity\|signal
					transduction
miR-155	AI806395	ALFY	ALFY	P + T	binding\|zinc ion binding
miR-155	NM_000038	APC	adenomatosis polyposis coli	P + T	Wnt receptor signaling pathway\|beta-
					catenin binding\|cell
					adhesion\|microtubule
					binding\|negative regulation of cell
					cycle\|protein\|complex
					assembly\|signal transduction
miR-155	NM_017610	ARK	Arkadia	P + T	protein ubiquitination\|ubiquitin
					ligase complex\|ubiquitin-protein
					ligase activity\|zinc ion binding
miR-155	BG032269	ARL8	ADP-ribosylation-like factor 8	M + P + T	GTP binding\|small GTPase mediated
					signal transduction
miR-155	AB000815	ARNTL	aryl hydrocarbon receptor	P + T	circadian rhythm\|nucleus\|regulation
			nuclear translocator-like		of transcription, DNA-
					dependent\|signal transducer
					activity\|signal
					transduction\|transcription\|transcription
					factor activity
miR-155	NM_001670	ARVCF	armadillo repeat gene	P + T	cell
			deletes in velocardiofacial		adhesion\|cytoskeleton\|development\|protein
			syndrome		binding\|structural molecule
					activity
miR-155	AK024064	ASTN2	astrotactin 2	P + T	integral to membrane
miR-155	M95541	ATP2B1	ATPase, Ca++ transporting,	M + P + T	ATP binding\|calcium ion
			plasma membrane
1		binding\|calcium ion
					transport\|calcium-transporting
					ATPase activity\|calmodulin
					binding\|cation transport\|hydrolase
					activity\|hydrolase activity, acting on
					acid anhydrides, catalyzing
					transmembrane movement of
					substances\|integral to plasma
					membrane\|magnesium ion
					binding\|membrane\|metabolism
miR-155	NM_001186	BACH1	BTB and CNC homology 1,	P + T	DNA binding\|nucleus\|protein
			basic leucine zipper		binding\|regulation of transcription,
			transcription factor 1		DNA-
					dependent\|transcription\|transcription
					factor activity
miR-155	NM_007005	BCE-1	BCE-1 protein	P + T	frizzled signaling
					pathway\|molecular_function
					unknown\|nucleus\|nucleus\|regulation
					of transcription\|regulation of
					transcription, DNA-dependent
miR-155	NM_022893	BCL11A	B-cell CLL/lymphoma 11A	P + T	cytoplasm\|hemopoiesis\|nucleic acid
			(zinc finger protein)		binding\|nucleus\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|zinc ion
					binding
miR-155	NM_001709	BDNF	brain-derived neurotrophic	M + T	growth factor activity\|growth factor
			factor		activity\|neurogenesis
miR-155	NM_014577	BRD1	bromodomain containing 1	P + T	DNA binding\|cell
					cycle\|nucleus\|nucleus\|regulation of
					transcription, DNA-dependent
miR-155	NM_024529	C1orf28	chromosome	1 open reading	M + P + T
			frame 28
miR-155	NM_000719	CACNA1C	calcium channel, voltage-	P + T	calcium ion binding\|calcium ion
			dependent, L type, alpha 1C		transport\|cation transport\|integral to
			subunit		membrane\|ion channel activity\|ion
					transport\|membrane\|regulation of
					heart contraction rate\|voltage-gated
					calcium channel activity\|voltage-
					gated calcium channel
					activity\|voltage-gated calcium
					channel complex\|voltage-gated
					calcium channel complex
miR-155	AL118798	CD47	CD47 antigen (Rh-related	P + T	cell-matrix adhesion\|integral to
			antigen, integrin-associated		plasma membrane\|integrin-mediated
			signal transducer)		signaling pathway\|plasma
					membrane\|protein binding
miR-155	AL564683	CEBPB	CCAAT/enhancer binding	M + P + T	acute-phase response\|inflammatory
			protein (C/EBP), beta		response\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					factor activity\|transcription from
					RNA polymerase II promoter
miR-155	NM_007023	CGEF2	cAMP-regulated guanine	M + P	3′,5′-cAMP binding\|G-protein
			nucleotide exchange factor		coupled receptor protein signaling
			II		pathway\|cAMP-dependent protein
					kinase complex\|cAMP-dependent
					protein kinase regulator
					activity\|exocytosis\|guanyl-nucleotide
					exchange factor activity\|membrane
					fraction\|nucleotide binding\|protein
					amino acid phosphorylation\|small
					GTPase mediated signal transduction
miR-155	AU152178	CMG2	capillary morphogenesis	P + T	integral to membrane\|receptor
			protein
2		activity
miR-155	NM_005776	CNIH	cornichon homolog	P + T	immune response\|integral to
			(Drosophila)		membrane\|intracellular signaling
					cascade\|membrane
miR-155	AW241703	CNTN4	Homo sapiens cDNA	P + T	cell adhesion\|membrane\|protein
			FLJ32716 fis, clone		binding
			TESTI2000808, highly
			similar to Rattus norvegicus
			neural cell adhesion protein
			BIG-2 precursor (BIG-2)
			mRNA, mRNA sequence
miR-155	NM_000094	COL7A1	collagen, type VII, alpha 1	P + T	basement membrane\|cell
			(epidermolysis bullosa,		adhesion\|collagen type
			dystrophic, dominant and		VII\|cytoplasm\|epidermis
			recessive)		development\|phosphate
					transport\|protein binding\|serine-type
					endopeptidase inhibitor
					activity\|structural molecule activity
miR-155	NM_003653	COPS3	COP9 constitutive	P + T	signalosome complex
			photomorphogenic homolog
			subunit 3 (Arabidopsis)
miR-155	NM_005211	CSF1R	colony stimulating factor 1	M + P + T	ATP binding\|antimicrobial humoral
			receptor, formerly		response (sensu Vertebrata)\|cell
			McDonough feline sarcoma		proliferation\|development\|integral to
			viral (v-fms) oncogene		plasma membrane\|macrophage
			homolog		colony stimulating factor receptor
					activity\|plasma membrane\|protein
					amino acid phosphorylation\|receptor
					activity\|signal
					transduction\|transferase
					activity\|transmembrane receptor
					protein tyrosine kinase signaling
					pathway
miR-155	NM_001892	CSNK1A1	casein kinase 1, alpha 1	P + T	ATP binding\|Wnt receptor signaling
					pathway\|casein kinase I
					activity\|protein amino acid
					phosphorylation\|protein amino acid
					phosphorylation\|protein
					serine/threonine kinase
					activity\|protein-tyrosine kinase
					activity\|transferase activity
miR-155	NM_005214	CTLA4	cytotoxic T-lymphocyte-	P + T	immune response\|immune
			associated protein 4		response\|integral to plasma
					membrane\|membrane
miR-155	U69546	CUGBP2	CUG triplet repeat, RNA	M + P + T	RNA binding\|RNA binding\|RNA
			binding protein
2		processing\|neuromuscular junction
					development\|nucleotide
					binding\|regulation of heart
					contraction rate
miR-155	NM_030927	DC-TM4F2	tetraspanin similar to	P + T	integral to membrane
			TM4SF9
miR-155	NM_015652	DKFZP564P1916	DKFZP564P1916 protein	P + T
miR-155	AF151831	DKFZP566C134	DKFZP566C134 protein	P + T	protein binding
miR-155	NM_004411	DNCI1	dynein, cytoplasmic,	P + T	cytoplasmic dynein complex\|motor
			intermediate polypeptide 1		activity
miR-155	NM_001400	EDG1	endothelial differentiation,	P + T	G-protein coupled receptor protein
			sphingolipid G-protein-		signaling pathway\|cell
			coupled receptor, 1		adhesion\|integral to plasma
					membrane\|lysosphingolipid and
					lysophosphatidic acid receptor
					activity\|plasma membrane\|receptor
					activity\|signal transduction
miR-155	NM_006795	EHD1	EH-domain containing 1	P + T	ATP binding\|GTP binding\|GTPase
					activity\|biological_process
					unknown\|calcium ion
					binding\|cellular_component
					unknown
miR-155	NM_012081	ELL2	ELL-related RNA	M + P + T	RNA elongation from RNA
			polymerase II, elongation		polymerase II promoter\|RNA
			factor		polymerase II transcription factor
					activity\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					elongation factor complex
miR-155	NM_005238	ETS1	v-ets erythroblastosis virus	P + T	RNA polymerase II transcription
			E26 oncogene homolog 1		factor activity\|immune
			(avian)		response\|negative regulation of cell
					proliferation\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					factor activity\|transcription from
					RNA polymerase II promoter
miR-155	NM_002009	FGF7	fibroblast growth factor 7	P + T	cell proliferation\|cell-cell
			(keratinocyte growth factor)		signaling\|epidermis
					development\|extracellular
					region\|growth factor activity\|positive
					regulation of cell
					proliferation\|regulation of cell
					cycle\|response to wounding\|signal
					transduction
miR-155	NM_018208	FLJ10761	hypothetical protein	P + T	biological_process
			FLJ10761		unknown\|cellular_component
					unknown\|choline kinase
					activity\|transferase activity
miR-155	NM_018243	FLJ10849	hypothetical protein	P + T	GTP binding\|cell cycle\|cytokinesis
			FLJ10849
miR-155	NM_022064	FLJ12565	hypothetical protein	P + T	ligase activity\|protein
			FLJ12565		ubiquitination\|ubiquitin ligase
					complex\|ubiquitin-protein ligase
					activity\|zinc ion binding
miR-155	NM_018391	FLJ23277	FLJ23277 protein	P + T
miR-155	NM_021078	GCN5L2	GCN5 general control of	M + P + T	N-acetyltransferase
			amino-acid synthesis 5-like		activity\|chromatin
			2 (yeast)		remodeling\|histone acetyltransferase
					activity\|histone deacetylase
					binding\|nucleus\|protein amino acid
					acetylation\|regulation of
					transcription from RNA polymerase II
					promoter\|transcription\|transcription
					coactivator activity\|transferase
					activity
miR-155	NM_018178	GPP34R	hypothetical protein	P + T
			FLJ10687
miR-155	AF019214	HBP1	HMG-box containing	M + P	DNA binding\|nucleus\|regulation of
			protein 1		transcription, DNA-dependent
miR-155	NM_006037	HDAC4	histone deacetylase 4	P + T	B-cell differentiation\|cell
					cycle\|chromatin
					modification\|cytoplasm\|develop-
					ment\|histone deacetylase activity\|histone
					deacetylase complex\|hydrolase
					activity\|inflammatory
					response\|negative regulation of
					myogenesis\|neurogenesis\|nucleus\|regulation
					of transcription, DNA-
					dependent\|transcription\|transcription
					factor binding\|transcriptional
					repressor activity
miR-155	NM_001530	HIF1A	hypoxia-inducible factor 1,	P + T	RNA polymerase II transcription
			alpha subunit (basic helix-		factor activity, enhancer
			loop-helix transcription		binding\|electron transport\|histone
			factor)		acetyltransferase
					binding\|homeostasis\|nucleus\|nu-
					cleus\|protein heterodimerization
					activity\|protein heterodimerization
					activity\|regulation of transcription,
					DNA-dependent\|response to
					hypoxia\|signal transducer
					activity\|signal transduction\|signal
					transduction\|transcription factor
					activity
miR-155	AL023584	HIVEP2	human immunodeficiency	P + T
			virus type I enhancer
			binding protein
2
miR-155	AI682088	HLCS	holocarboxylase synthetase	P + T	biotin-[acetyl-CoA-carboxylase]
			(biotin-[proprionyl-		ligase activity\|biotin-
			Coenzyme A-carboxylase		[methylcrotonoyl-CoA-carboxylase]
			(ATP-hydrolysing)] ligase)		ligase activity\|biotin-
					[methylmalonyl-CoA-
					carboxytransferase] ligase
					activity\|biotin-[propionyl-CoA-
					carboxylase (ATP-hydrolyzing)]
					ligase activity\|ligase activity\|protein
					modification
miR-155	NM_020190	HNOEL-iso	HNOEL-iso protein	P + T
miR-155	NM_014002	IKBKE	inhibitor of kappa light	P + T	ATP binding\|NF-kappaB-inducing
			polypeptide gene enhancer		kinase activity\|cytoplasm\|immune
			in B-cells, kinase epsilon		response\|positive regulation of I-
					kappaB kinase/NF-kappaB
					cascade\|protein amino acid
					phosphorylation\|protein
					serine/threonine kinase
					activity\|signal transducer
					activity\|transferase activity
miR-155	D13720	ITK	IL2-inducible T-cell kinase	P + T	ATP binding\|cellular defense
					response\|intracellular signaling
					cascade\|non-membrane spanning
					protein tyrosine kinase
					activity\|protein amino acid
					phosphorylation\|transferase activity
miR-155	NM_002249	KCNN3	potassium	P + T	calcium-activated potassium channel
			intermediate/small		activity\|calcium-activated potassium
			conductance calcium-		channel activity\|calmodulin
			activated channel, subfamily		binding\|integral to membrane\|ion
			N, member 3		channel activity\|ion
					transport\|membrane\|membrane
					fraction\|neurogenesis\|potassium ion
					transport\|potassium ion
					transport\|small conductance calcium-
					activated potassium channel
					activity\|synaptic
					transmission\|voltage-gated potassium
					channel complex
miR-155	AB033100	KIAA1274	KIAA protein (similar to	P + T	protein tyrosine phosphatase activity
			mouse paladin)
miR-155	NM_017780	KIAA1416	KIAA1416 protein	P + T	ATP binding\|chromatin\|chromatin
					assembly or disassembly\|chromatin
					binding\|helicase activity\|nucleus
miR-155	NM_002264	KPNA1	karyopherin alpha 1	P + T	NLS-bearing substrate-nucleus
			(importin alpha 5)		import\|cytoplasm\|intracellular
					protein transport\|nuclear localization
					sequence binding\|nuclear
					pore\|nucleus\|protein binding\|protein
					transporter activity\|regulation of
					DNA recombination
miR-155	AK021602	KPNA4	karyopherin alpha 4	P + T	NLS-bearing substrate-nucleus
			(importin alpha 3)		import\|binding\|intracellular protein
					transport\|nucleus\|protein transporter
					activity
miR-155	NM_020354	LALP1	lysosomal apyrase-like	M + P + T	hydrolase activity
			protein
1
miR-155	AW242408	LOC151531	Similar to uridine	M + P + T	cytosol\|nucleoside
			phosphorylase		metabolism\|nucleotide
			[Homo sapiens],		catabolism\|protein
			mRNA sequence		binding\|transferase activity,
					transferring glycosyl groups\|type III
					intermediate filament\|uridine
					metabolism\|uridine phosphorylase
					activity
miR-155	NM_016210	LOC51161	g20 protein	P + T
miR-155	NM_018557	LRP1B	low density lipoprotein-	P + T	calcium ion binding\|integral to
			related protein 1B (deleted		membrane\|low-density lipoprotein
			in tumors)		receptor activity\|membrane\|protein
					transport\|receptor activity\|receptor
					mediated endocytosis
miR-155	NM_002446	MAP3K10	mitogen-activated protein	M + P + T	ATP binding\|JUN kinase kinase
			kinase kinase kinase 10		kinase activity\|activation of
					JNK\|autophosphorylation\|induction
					of apoptosis\|protein
					homodimerization activity\|protein
					serine/threonine kinase
					activity\|protein-tyrosine kinase
					activity\|signal
					transduction\|transferase activity
miR-155	NM_003954	MAP3K14	mitogen-activated protein	P + T	ATP binding\|protein amino acid
			kinase kinase kinase 14		phosphorylation\|protein
					serine/threonine kinase
					activity\|transferase activity
miR-155	AL117407	MAP3K7IP2	mitogen-activated protein	P + T	kinase activity\|positive regulation of
			kinase kinase kinase 7		I-kappaB kinase/NF-kappaB
			interacting protein
2		cascade\|positive regulation of I-
					kappaB kinase/NF-kappaB
					cascade\|signal transducer
					activity\|signal transducer activity
miR-155	NM_004992	MECP2	methyl CpG binding protein	M + P + T	DNA binding\|negative regulation of
			2 (Rett syndrome)		transcription from RNA polymerase
					II promoter\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					corepressor activity
miR-155	NM_002398	MEIS1	Meis1, myeloid ecotropic	M + P + T	RNA polymerase II transcription
			viral integration site 1		factor activity\|nucleus\|regulation of
			homolog (mouse)		transcription, DNA-
					dependent\|transcription factor
					activity
miR-155	NM_016289	MO25	MO25 protein	P + T
miR-155	AA621962	MYO1D	myosin ID	M + P + T	ATP binding\|actin
					binding\|calmodulin binding\|motor
					activity\|myosin
miR-155	NM_030571	N4WBP5	likely ortholog of mouse	P + T	positive regulation of I-kappaB
			Nedd4 WW binding protein 5		kinase/NF-kappaB cascade\|signal
					transducer activity
miR-155	NM_014903	NAV3	neuron navigator 3	P + T	ATP
					binding\|mitochondrion\|nucleoside-
					triphosphatase activity\|nucleotide
					binding
miR-155	NM_030571	NDFIP1	likely ortholog of mouse	P + T	positive regulation of I-kappaB
			Nedd4 WW binding protein 5		kinase/NF-kappaB cascade\|signal
					transducer activity
miR-155	NM_006599	NFAT5	nuclear factor of activated	M + P + T	RNA polymerase II transcription
			T-cells 5, tonicity-		factor
			responsive		activity\|excretion\|nucleus\|regulation
					of transcription, DNA-
					dependent\|signal
					transduction\|transcription factor
					activity\|transcription from RNA
					polymerase II promoter
miR-155	NM_002515	NOVA1	neuro-oncological ventral	M + P + T	RNA binding\|RNA binding\|RNA
			antigen
1		splicing\|RNA splicing\|locomotory
					behavior\|locomotory
					behavior\|nucleus\|synaptic
					transmission\|synaptic transmission
miR-155	AI373299	PANK1	pantothenate kinase 1	P + T	ATP binding\|coenzyme A
					biosynthesis\|pantothenate kinase
					activity\|transferase activity
miR-155	BG110231	PAPOLA	poly(A) polymerase alpha	P + T	RNA binding\|cytoplasm\|mRNA
					polyadenylylation\|mRNA
					processing\|nucleus\|polynucleotide
					adenylyltransferase
					activity\|transcription\|transferase
					activity
miR-155	NM_020403	PCDH9	protocadherin 9	M + P + T	calcium ion binding\|cell
					adhesion\|homophilic cell
					adhesion\|integral to
					membrane\|membrane\|protein binding
miR-155	NM_002655	PLAG1	pleiomorphic adenoma gene 1	P + T	nucleic acid
					binding\|nucleus\|transcription factor
					activity\|zinc ion binding
miR-155	AJ272212	PSKH1	protein serine kinase H1	P + T	ATP binding\|Golgi
					apparatus\|nucleus\|protein amino acid
					phosphorylation\|protein
					serine/threonine kinase
					activity\|transferase activity
miR-155	NM_014904	Rab11-FIP2	KIAA0941 protein	P + T
miR-155	AF322067	RAB34	RAB34, member RAS	P + T	GTP binding\|Golgi apparatus\|protein
			oncogene family		transport\|small GTPase mediated
					signal transduction
miR-155	NM_002869	RAB6A	RAB6A, member RAS	M + P + T	GTP binding\|GTPase activity\|Golgi
			oncogene family		apparatus\|protein transport\|small
					GTPase mediated signal transduction
miR-155	AL136727	RAB6C	RAB6C, member RAS	M + P + T	GTP binding\|GTPase
			oncogene family		activity\|intracellular\|protein
					transport\|response to drug\|small
					GTPase mediated signal transduction
miR-155	NM_002902	RCN2	reticulocalbin	2, EF-hand	P + T	calcium ion binding\|endoplasmic
			calcium binding domain		reticulum\|protein binding
miR-155	AJ223321	RP58	zinc finger protein 238	M + P + T
miR-155	NM_002968	SALL1	sal-like 1 (Drosophila)	P + T	morphogenesis\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					factor activity\|zinc ion binding
miR-155	NM_002971	SATB1	special AT-rich sequence	P + T	double-stranded DNA
			binding protein 1 (binds to		binding\|establishment and/or
			nuclear matrix/scaffold-		maintenance of chromatin
			associating DNA's)		architecture\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription factor
					activity
miR-155	NM_003469	SCG2	secretogranin II	P + T	calcium ion binding\|protein secretion
			(chromogranin C)
miR-155	NM_005625	SDCBP	syndecan binding protein	P + T	actin cytoskeleton organization and
			(syntenin)		biogenesis\|adherens
					junction\|cytoskeletal adaptor
					activity\|cytoskeleton\|endoplasmic
					reticulum\|interleukin-5 receptor
					binding\|interleukin-5 receptor
					complex\|intracellular signaling
					cascade\|metabolism\|neurexin
					binding\|nucleus\|oxidoreductase
					activity\|plasma membrane\|protein
					binding\|protein heterodimerization
					activity\|protein-membrane
					targeting\|substrate-bound cell
					migration, cell extension\|synaptic
					transmission\|syndecan binding
miR-155	NM_000232	SGCB	sarcoglycan, beta (43 kDa	P + T	cytoskeleton\|cytoskeleton
			dystrophin-associated		organization and biogenesis\|integral
			glycoprotein)		to plasma membrane\|muscle
					development\|sarcoglycan complex
miR-155	NM_013257	SGKL	serum/glucocorticoid	P + T	ATP binding\|intracellular signaling
			regulated kinase-like		cascade\|protein amino acid
					phosphorylation\|protein amino acid
					phosphorylation\|protein
					serine/threonine kinase
					activity\|protein serine/threonine
					kinase activity\|protein-tyrosine
					kinase activity\|response to
					stress\|transferase activity
miR-155	NM_005069	SIM2	single-minded homolog 2	P + T	cell
			(Drosophila)		differentiation\|neurogenesis\|nucleus\|regulation
					of transcription, DNA-
					dependent\|signal transducer
					activity\|signal
					transduction\|transcription\|transcription
					factor activity
miR-155	AA927480	SKI	v-ski sarcoma viral	P + T
			oncogene homolog (avian)
miR-155	NM_006748	SLA	Src-like-adaptor	P + T	SH3/SH2 adaptor
					activity\|intracellular signaling
					cascade
miR-155	AI684141	SMARCA4	SWI/SNF related, matrix	P + T	ATP binding\|DNA binding\|helicase
			associated, actin dependent		activity\|helicase activity\|hydrolase
			regulator of chromatin,		activity\|nucleoplasm\|nucleus\|regulation
			subfamily a, member 4		of transcription from RNA
					polymerase II
					promoter\|transcription\|transcription
					coactivator activity\|transcription
					factor activity
miR-155	AB005043	SOCS1	suppressor of cytokine	M + P + T	JAK-STAT
			signaling
1		cascade\|cytoplasm\|insulin-like
					growth factor receptor
					binding\|intracellular signaling
					cascade\|negative regulation of JAK-
					STAT cascade\|protein kinase
					binding\|protein kinase inhibitor
					activity\|regulation of cell
					growth\|ubiquitin cycle
miR-155	NM_004232	SOCS4	suppressor of cytokine	M + P	JAK-STAT
			signaling 4		cascade\|cytoplasm\|defense
					response\|intracellular signaling
					cascade\|regulation of cell growth
miR-155	NM_005986	SOX1	SRY (sex determining	P + T	DNA binding\|establishment and/or
			region Y)-box 1		maintenance of chromatin
					architecture\|nucleus\|regulation of
					transcription, DNA-
					dependent\|regulation of transcription,
					DNA-dependent\|transcription factor
					activity
miR-155	AI360875	SOX11	SRY (sex determining	M + T	DNA
			region Y)-box 11		binding\|neurogenesis\|nucleus\|regulation
					of transcription, DNA-
					dependent\|transcription
miR-155	AL136780	SOX6	SRY (sex determining	P + T	establishment and/or maintenance of
			region Y)-box 6		chromatin architecture\|heart
					development\|muscle
					development\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					factor activity
miR-155	AW470841	SP3	Sp3 transcription factor	P + T	DNA binding\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcriptional
					activator activity\|transcriptional
					repressor activity\|zinc ion binding
miR-155	BF224259	SPF30	splicing factor 30, survival	P + T	RNA splicing\|RNA splicing factor
			of motor neuron-related		activity, transesterification
					mechanism\|apoptosis\|cytoplasm\|induction
					of apoptosis\|spliceosome
					assembly\|spliceosome complex
miR-155	NM_003120	SPI1	spleen focus forming virus	M + T	negative regulation of transcription
			(SFFV) proviral integration		from RNA polymerase II
			oncogene spi1		promoter\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					factor activity
miR-155	BE676214	SSH2	slingshot 2	P + T	protein amino acid
					dephosphorylation\|protein
					tyrosine/serine/threonine
					phosphatase activity
miR-155	AF159447	SUFU	suppressor of fused homolog	P + T	cell
			(Drosophila)		cycle\|cytoplasm\|development\|negative
					regulation of cell
					cycle\|nucleus\|proteolysis and
					peptidolysis\|signal transducer
					activity\|signal transduction\|skeletal
					development\|transcription
					corepressor activity
miR-155	NM_006754	SYPL	synaptophysin-like protein	M + P + T	integral to plasma
					membrane\|membrane\|synaptic
					transmission\|synaptic
					vesicle\|transport\|transporter activity
miR-155	NM_006286	TFDP2	transcription factor Dp-2	P + T	DNA metabolism\|nucleus\|regulation
			(E2F dimerization partner 2)		of cell cycle\|regulation of
					transcription from RNA polymerase II
					promoter\|transcription\|transcription
					cofactor activity\|transcription factor
					activity\|transcription factor complex
miR-155	AA705845	TLE4	transducin-like enhancer of	P + T	frizzled signaling
			split 4 (E(sp1) homolog,		pathway\|molecular_function
			Drosophila)		unknown\|nucleus\|nucleus\|regulation
					of transcription\|regulation of
					transcription, DNA-dependent
miR-155	NM_014765	TOMM20	translocase of outer	P + T	integral to membrane\|mitochondrial
			mitochondrial membrane 20		outer membrane translocase
			(yeast) homolog		complex\|mitochondrion\|outer
					membrane\|protein translocase
					activity\|protein-mitochondrial
					targeting
miR-155	AW341649	TP53INP1	tumor protein p53 inducible	P + T	apoptosis\|nucleus
			nuclear protein 1
miR-155	BC005016	TRIM2	tripartite motif-containing 2	P + T	cytoplasm\|myosin binding\|protein
					ubiquitination\|ubiquitin ligase
					complex\|ubiquitin-protein ligase
					activity\|zinc ion binding
miR-155	AA524505	TSGA	zinc finger protein	P + T	nucleus
miR-155	AW157525	TSGA14	testis specific, 14	M + P + T	centrosome
miR-155	X62048	WEE1	WEE1 homolog (S. pombe)	P + T	ATP
					binding\|cytokinesis\|mitosis\|nucleus\|protein
					amino acid
					phosphorylation\|protein
					serine/threonine kinase
					activity\|protein-tyrosine kinase
					activity\|regulation of cell
					cycle\|transferase activity
miR-155	AC005539	WUGSC:	Similar to uridine	M + P + T
		H_NH0335J18.1	phosphorylase
			[Homo sapiens],
			mRNA sequence
miR-155	NM_003413	ZIC3	Zic family member 3	P + T	DNA binding\|determination of
			heterotaxy 1 (odd-paired		left/right
			homolog, Drosophila)		symmetry\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|zinc ion
					binding
miR-155	NM_007345	ZNF236	zinc finger protein 236	P + T	nucleus\|regulation of transcription,
					DNA-
					dependent\|transcription\|transcription
					factor activity\|zinc ion binding
miR-155	NM_006352	ZNF238	zinc finger protein 238	M + P + T	chromosome organization and
					biogenesis (sensu
					Eukaryota)\|negative regulation of
					transcription from RNA polymerase
					II promoter\|nuclear
					chromosome\|nucleic acid
					binding\|nucleus\|protein
					binding\|protein binding\|regulation of
					transcription, DNA-
					dependent\|specific RNA polymerase
					II transcription factor
					activity\|transcription\|transcription
					factor activity\|transport\|zinc ion
					binding
miR-21	NM_005164	ABCD2	ATP-binding cassette, sub-	M + P	ATP binding\|ATP-binding cassette
			family D (ALD), member 2		(ABC) transporter complex\|ATPase
					activity\|ATPase activity, coupled to
					transmembrane movement of
					substances\|fatty acid
					metabolism\|integral to plasma
					membrane\|membrane\|peroxisome\|transport
miR-21	NM_001616	ACVR2	activin A receptor, type II	P + T	ATP binding\|integral to plasma
					membrane\|membrane\|protein amino
					acid phosphorylation\|receptor
					activity\|transferase
					activity\|transforming growth factor
					beta receptor activity\|transmembrane
					receptor protein serine/threonine
					kinase signaling pathway
miR-21	NM_015339	ADNP	activity-dependent	P + T	nucleus\|regulation of transcription,
			neuroprotector		DNA-dependent\|transcription factor
					activity\|zinc ion binding
miR-21	AI990366	ARHGEF7	Rho guanine nucleotide	P + T	guanyl-nucleotide exchange factor
			exchange factor (GEF) 7		activity\|signal transduction
miR-21	NM_017610	ARK	Arkadia	P + T	protein ubiquitination\|ubiquitin
					ligase complex\|ubiquitin-protein
					ligase activity\|zinc ion binding
miR-21	NM_014034	ASF1A	DKFZP547E2110 protein	P + T	chromatin binding\|loss of chromatin
					silencing\|nucleus
miR-21	NM_017680	ASPN	asporin (LRR class 1)	P + T
miR-21	NM_000657	BCL2	B-cell CLL/lymphoma 2	P + T	anti-apoptosis\|endoplasmic
					reticulum\|humoral immune
					response\|integral to
					membrane\|membrane\|mitochondrial
					outer membrane\|mitochondrial outer
					membrane\|mitochondrion\|negative
					regulation of cell
					proliferation\|nucleus\|protein
					binding\|regulation of
					apoptosis\|regulation of cell
					cycle\|release of cytochrome c from
					mitochondria
miR-21	NM_014577	BRD1	bromodomain containing 1	P + T	DNA binding\|cell
					cycle\|nucleus\|nucleus\|regulation of
					transcription, DNA-dependent
miR-21	AA902767	BRD2	bromodomain containing 2	P + T	nucleus\|protein serine/threonine
					kinase activity\|spermatogenesis
miR-21	NM_014962	BTBD3	BTB (POZ) domain	P + T	protein binding
			containing 3
miR-21	NM_006763	BTG2	BTG family, member 2	P + T	DNA repair\|negative regulation of
					cell proliferation\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					factor activity
miR-21	AK025768	C20orf99	chromosome	20 open	P + T	nucleic acid binding
			reading frame 99
miR-21	AI671238	CAPN3	Homo sapiens cDNA	P + T	calcium ion binding\|calpain
			FLJ23750 fis, clone		activity\|calpain
			HEP16527, mRNA		activity\|intracellular\|intracellular\|muscle
			sequence		development\|proteolysis and
					peptidolysis\|proteolysis and
					peptidolysis\|signal transducer
					activity
miR-21	NM_002981	CCL1	chemokine (C-C motif)	P + T	calcium ion homeostasis\|cell-cell
			ligand
1		signaling\|chemokine
					activity\|chemotaxis\|extracellular
					space\|inflammatory response\|sensory
					perception\|signal transduction\|viral
					life cycle
miR-21	BF939071	CCM1	cerebral cavernous	M + P	binding\|catalytic
			malformations
1		activity\|cytoskeleton\|small GTPase
					mediated signal transduction\|small
					GTPase regulator activity
miR-21	NM_001789	CDC25A	cell division cycle 25A	P + T	cell
					proliferation\|cytokinesis\|hydrolase
					activity\|intracellular\|mitosis\|protein
					amino acid
					dephosphorylation\|protein tyrosine
					phosphatase activity\|regulation of
					cyclin dependent protein kinase
					activity
miR-21	NM_001842	CNTFR	ciliary neurotrophic factor	M + P + T	ciliary neurotrophic factor receptor
			receptor		activity\|cytokine binding\|extrinsic to
					membrane\|neurogenesis\|receptor
					activity\|signal transduction
miR-21	NM_001310	CREBL2	cAMP responsive element	P + T	nucleus\|regulation of transcription,
			binding protein-like 2		DNA-dependent\|signal
					transduction\|transcription\|transcription
					factor activity
miR-21	NM_016441	CRIM1	cysteine-rich motor neuron 1	M + P + T	insulin-like growth factor receptor
					activity\|integral to
					membrane\|membrane
					fraction\|neurogenesis\|serine-type
					endopeptidase inhibitor activity
miR-21	NM_015396	DKFZP434A043	DKFZP434A043 protein	P + T	cell adhesion\|cytoskeleton\|mitotic
					chromosome condensation\|protein
					binding\|structural molecule activity
miR-21	AL047650	DKFZp434A2417	endozepine-related protein	P + T	acyl-CoA binding
			precursor
miR-21	AB028628	DKFZP547E2110	DKFZP547E2110 protein	P + T	chromatin binding\|loss of chromatin
					silencing\|nucleus
miR-21	NM_031305	DKFZP564B1162	hypothetical protein	P + T	GTPase activator activity
			DKFZp564B1162
miR-21	NM_004405	DLX2	distal-less homeo box 2	P + T	brain
					development\|development\|nucleus\|regulation
					of transcription, DNA-
					dependent\|transcription factor
					activity
miR-21	NM_001949	E2F3	E2F transcription factor 3	M + P + T	nucleus\|protein binding\|regulation of
					cell cycle\|regulation of transcription,
					DNA-
					dependent\|transcription\|transcription
					factor activity\|transcription factor
					complex\|transcription initiation from
					RNA polymerase II promoter
miR-21	NM_006795	EHD1	EH-domain containing 1	P + T	ATP binding\|GTP binding\|GTPase
					activity\|biological_process
					unknown\|calcium ion
					binding\|cellular_component
					unknown
miR-21	NM_001412	EIF1A	eukaryotic translation	P + T	RNA binding\|eukaryotic translation
			initiation factor 1A		initiation factor 4F complex\|protein
					biosynthesis\|translation initiation
					factor activity\|translational
					initiation\|translational initiation
miR-21	AI832074	EIF2C2	eukaryotic translation	P + T	cellular_component unknown\|protein
			initiation factor
2C, 2		biosynthesis\|translation initiation
					factor activity
miR-21	NM_006874	ELF2	E74-like factor 2 (ets	P + T	nucleus\|nucleus\|protein
			domain transcription factor)		binding\|protein binding\|regulation of
					transcription from RNA polymerase
					II promoter\|regulation of
					transcription, DNA-
					dependent\|transcription factor
					activity\|transcriptional activator
					activity\|transcriptional activator
					activity
miR-21	NM_004438	EPHA4	EphA4	P + T	ATP binding\|ephrin receptor
					activity\|integral to plasma
					membrane\|membrane\|protein amino
					acid phosphorylation\|receptor
					activity\|signal
					transduction\|transferase
					activity\|transmembrane receptor
					protein tyrosine kinase signaling
					pathway
miR-21	BE888593	FLJ11220	hypothetical protein	P + T
			FLJ11220
miR-21	NM_017637	FLJ20043	hypothetical protein	P + T	nucleic acid binding\|nucleus\|zinc ion
			FLJ20043		binding
miR-21	AF019214	HBP1	HMG-box containing	M + P + T	DNA binding\|nucleus\|regulation of
			protein 1		transcription, DNA-dependent
miR-21	NM_000214	JAG1	jagged 1 (Alagille	M + P + T	Notch binding\|Notch signaling
			syndrome)		pathway\|angiogenesis\|calcium ion
					binding\|calcium ion binding\|cell
					communication\|cell fate
					determination\|development\|endothelial
					cell differentiation\|extracellular
					region\|growth factor
					activity\|hemopoiesis\|integral to
					plasma membrane\|keratinocyte
					differentiation\|membrane\|myoblast
					differentiation\|neurogenesis\|regulation
					of cell migration\|regulation of cell
					proliferation\|structural molecule
					activity
miR-21	NM_002232	KCNA3	potassium voltage-gated	M + P + T	cation transport\|delayed rectifier
			channel, shaker-related		potassium channel activity\|integral to
			subfamily, member 3		membrane\|membrane\|membrane
					fraction\|potassium ion
					transport\|voltage-gated potassium
					channel complex
miR-21	NM_014766	KIAA0193	KIAA0193 gene product	P + T	cellular_component
					unknown\|dipeptidase
					activity\|exocytosis\|proteolysis and
					peptidolysis
miR-21	NM_014912	KIAA0940	KIAA0940 protein	M + P + T	nucleic acid binding
miR-21	NM_014952	KIAA0945	KIAA0945 protein	P + T	DNA binding
miR-21	NM_017780	KIAA1416	KIAA1416 protein	P + T	ATP binding\|chromatin\|chromatin
					assembly or disassembly\|chromatin
					binding\|helicase activity\|nucleus
miR-21	AB040901	KIAA1468	KIAA1468 protein	P + T	binding\|mitotic chromosome
					condensation
miR-21	U90268	Krit1	cerebral cavernous	M + P	binding\|catalytic
			malformations
1		activity\|cytoskeleton\|small GTPase
					mediated signal transduction\|small
					GTPase regulator activity
miR-21	BF591611	LOC147632	hypothetical protein	P + T	oxidoreductase activity\|zinc ion
			BC010734		binding
miR-21	NM_005904	MADH7	MAD, mothers against	P + T	intracellular\|protein binding\|receptor
			decapentaplegic homolog 7		signaling protein serine/threonine
			(Drosophila)		kinase signaling protein
					activity\|regulation of transcription,
					DNA-dependent\|response to
					stress\|transcription\|transforming
					growth factor beta receptor signaling
					pathway\|transforming growth factor
					beta receptor, inhibitory cytoplasmic
					mediator activity
miR-21	NM_025146	MAK3P	likely ortholog of mouse	P + T	N-acetyltransferase activity
			Mak3p homolog
			(S. cerevisiae)
miR-21	NM_014319	MAN1	integral inner nuclear	P + T	integral to membrane\|integral to
			membrane protein		nuclear inner membrane\|membrane
					fraction\|nuclear
					membrane\|nucleotide binding
miR-21	AW025150	MAP3K12	mitogen-activated protein	M + T	ATP binding\|JNK
			kinase kinase kinase 12		cascade\|cytoplasm\|magnesium ion
					binding\|plasma membrane\|protein
					amino acid phosphorylation\|protein
					kinase cascade\|protein
					serine/threonine kinase
					activity\|protein-tyrosine kinase
					activity\|transferase activity
miR-21	NM_012325	MAPRE1	microtubule-associated	P + T	cell
			protein, RP/EB family,		proliferation\|cytokinesis\|microtubule
			member
1		binding\|mitosis\|protein C-terminus
					binding\|regulation of cell cycle
miR-21	NM_002380	MATN2	matrilin 2	P + T	biological_process unknown\|calcium
					ion binding\|extracellular matrix
					(sensu Metazoa)
miR-21	NM_018834	MATR3	matrin 3	M + P + T	RNA binding\|nuclear inner
					membrane\|nucleotide
					binding\|nucleus\|structural molecule
					activity\|zinc ion binding
miR-21	NM_021038	MBNL1	muscleblind-like	M + P + T	cytoplasm\|double-stranded RNA
			(Drosophila)		binding\|embryonic development
					(sensu Mammalia)\|embryonic limb
					morphogenesis\|muscle
					development\|myoblast
					differentiation\|neurogenesis\|nucleic
					acid binding\|nucleus\|nucleus
miR-21	AI139252	MGC16063	ribosomal protein L35a	P + T	JAK-STAT cascade\|acute-phase
					response\|calcium ion binding\|cell
					motility\|cytoplasm\|hematopoietin/interferon-
					class (D200-domain)
					cytokine receptor signal transducer
					activity\|intracellular signaling
					cascade\|negative regulation of
					transcription from RNA polymerase II
					promoter\|neurogenesis\|nucleus\|nu-
					cleus\|regulation of transcription, DNA-
					dependent\|signal transducer
					activity\|transcription\|transcription
					factor activity\|transcription factor
					activity
miR-21	BC004162	MGC2452	hypothetical protein	P + T	fatty acid metabolism\|generation of
			MGC2452		precursor metabolites and
					energy\|ligand-dependent nuclear
					receptor activity\|lipid
					metabolism\|nucleus\|nucleus\|regulation
					of transcription, DNA-
					dependent\|steroid hormone receptor
					activity\|transcription\|transcription
					factor activity\|transcription factor
					activity\|transcription from RNA
					polymerase II promoter
miR-21	NM_024052	MGC3048	hypothetical protein	P + T
			MGC3048
miR-21	AB049636	MRPL9	mitochondrial ribosomal	P + T	mitochondrion\|protein
			protein L9		biosynthesis\|ribosome\|structural
					constituent of ribosome
miR-21	NM_015678	NBEA	neurobeachin	P + T	Golgi trans
					face\|cytosol\|endomembrane
					system\|plasma membrane\|post-Golgi
					transport\|postsynaptic
					membrane\|protein kinase A binding
miR-21	AI700518	NFIB	nuclear factor I/B	M + T	DNA
					replication\|nucleus\|nucleus\|regulation
					of transcription, DNA-
					dependent\|transcription\|transcription
					factor activity\|transcription factor
					activity
miR-21	NM_002527	NTF3	neurotrophin 3	M + P	anti-apoptosis\|cell motility\|cell-cell
					signaling\|growth factor
					activity\|neurogenesis\|signal
					transduction
miR-21	U24223	PCBP1	poly(rC) binding protein 1	M + P + T	RNA binding\|catalytic
					activity\|cytoplasm\|mRNA
					metabolism\|nucleus\|ribonucleoprotein
					complex\|single-stranded DNA
					binding
miR-21	NM_005016	PCBP2	poly(rC) binding protein 2	M + T	DNA binding\|RNA
					binding\|cytoplasm\|mRNA
					metabolism\|nucleic acid
					binding\|nucleus\|ribonucleoprotein
					complex
miR-21	NM_014456	PDCD4	programmed cell death 4	P + T	apoptosis
			(neoplastic transformation
			inhibitor)
miR-21	AF338650	PDZD2	PDZ domain containing 2	P + T
miR-21	NM_000325	PITX2	paired-like homeodomain	M + P + T	determination of left/right
			transcription factor
2		symmetry\|development\|nucleus\|organo-
					genesis\|regulation of transcription,
					DNA-dependent\|transcription factor
					activity
miR-21	NM_002655	PLAG1	pleiomorphic adenoma gene 1	P + T	nucleic acid
					binding\|nucleus\|transcription factor
					activity\|zinc ion binding
miR-21	NM_005036	PPARA	peroxisome proliferative	P + T	fatty acid metabolism\|generation of
			activated receptor, alpha		precursor metabolites and
					energy\|ligand-dependent nuclear
					receptor activity\|lipid
					metabolism\|nucleus\|nucleus\|regulation
					of transcription, DNA-
					dependent\|steroid hormone receptor
					activity\|transcription\|transcription
					factor activity\|transcription factor
					activity\|transcription from RNA
					polymerase II promoter
miR-21	NM_002711	PPP1R3A	protein phosphatase	1,	P + T	carbohydrate metabolism\|glycogen
			regulatory (inhibitor)		metabolism\|hydrolase
			subunit 3A (glycogen and		activity\|integral to
			sarcoplasmic reticulum		membrane\|phosphoprotein
			binding subunit, skeletal		phosphatase activity\|type 1
			muscle)		serine/threonine specific protein
					phosphatase inhibitor activity
miR-21	NM_000944	PPP3CA	protein phosphatase 3	P + T	calcineurin complex\|calcium ion
			(formerly 2B), catalytic		binding\|calmodulin
			subunit, alpha isoform		binding\|hydrolase activity\|protein
			(calcineurin A alpha)		amino acid
					dephosphorylation\|protein
					serine/threonine phosphatase activity
miR-21	NM_018569	PRO0971	hypothetical protein	P + T
			PRO0971
miR-21	AA156948	PRPF4B	PRP4 pre-mRNA processing	M + T	ATP binding\|RNA splicing\|nuclear
			factor 4 homolog B (yeast)		mRNA splicing, via
					spliceosome\|nucleus\|protein amino
					acid phosphorylation\|protein
					serine/threonine kinase
					activity\|transferase activity
miR-21	BF337790	PURB	purine-rich element binding	M + P + T
			protein B
miR-21	NM_002869	RAB6A	RAB6A, member RAS	P + T	GTP binding\|GTPase activity\|Golgi
			oncogene family		apparatus\|protein transport\|small
					GTPase mediated signal transduction
miR-21	AL136727	RAB6C	RAB6C, member RAS	P + T	GTP binding\|GTPase
			oncogene family		activity\|intracellular\|protein
					transport\|response to drug\|small
					GTPase mediated signal transduction
miR-21	NM_002890	RASA1	RAS p21 protein activator	P + T	Ras GTPase activator
			(GTPase activating protein) 1		activity\|intracellular signaling
					cascade
miR-21	NM_005739	RASGRP1	RAS guanyl releasing	P + T	Ras guanyl-nucleotide exchange
			protein 1 (calcium and		factor activity\|Ras protein signal
			DAG-regulated)		transduction\|calcium ion
					binding\|calcium ion
					binding\|diacylglycerol
					binding\|guanyl-nucleotide exchange
					factor activity\|membrane
					fraction\|small GTPase mediated
					signal transduction
miR-21	NM_021111	RECK	reversion-inducing-cysteine-	M + P + T	cell cycle\|membrane\|membrane
			rich protein with kazal		fraction\|metalloendopeptidase
			motifs		inhibitor activity\|negative regulation
					of cell cycle\|serine-type
					endopeptidase inhibitor activity
miR-21	NM_006915	RP2	retinitis pigmentosa 2 (X-	P + T	beta-tubulin
			linked recessive)		folding\|membrane\|sensory
					perception\|unfolded protein
					binding\|visual perception
miR-21	AA906056	RPS6KA3	ribosomal protein S6 kinase,	M + T	ATP binding\|central nervous system
			90 kDa, polypeptide 3		development\|protein amino acid
					phosphorylation\|protein
					serine/threonine kinase
					activity\|signal transduction\|skeletal
					development\|transferase activity
miR-21	NM_002971	SATB1	special AT-rich sequence	M + P + T	double-stranded DNA
			binding protein 1 (binds to		binding\|establishment and/or
			nuclear matrix/scaffold-		maintenance of chromatin
			associating DNA's)		architecture\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription factor
					activity
miR-21	NM_014191	SCN8A	sodium channel, voltage	M + P + T	ATP binding\|cation channel
			gated, type VIII, alpha		activity\|cation transport\|integral to
			polypeptide		membrane\|membrane\|neurogenesis\|sodium
					ion transport\|voltage-gated
					sodium channel activity\|voltage-
					gated sodium channel complex
miR-21	AA927480	SKI	v-ski sarcoma viral	M + P + T
			oncogene homolog (avian)
miR-21	NM_003983	SLC7A6	solute carrier family 7	P + T	amino acid metabolism\|amino acid
			(cationic amino acid		transport\|amino acid-polyamine
			transporter, y + system),		transporter activity\|integral to plasma
			member
6		membrane\|plasma membrane\|protein
					complex assembly\|transport
miR-21	NM_006359	SLC9A6	solute carrier family 9	P + T	antiporter activity\|endoplasmic
			(sodium/hydrogen		reticulum membrane\|integral to
			exchanger), isoform 6		membrane\|integral to membrane\|ion
					transport\|microsome\|mitochondrion\|regulation
					of pH\|sodium ion
					transport\|sodium:hydrogen antiporter
					activity\|solute:hydrogen antiporter
					activity
miR-21	NM_003076	SMARCD1	SWI/SNF related, matrix	P + T	chromatin remodeling\|chromatin
			associated, actin dependent		remodeling complex\|regulation of
			regulator of chromatin,		transcription from RNA polymerase
			subfamily d, member 1		II promoter\|transcription coactivator
					activity
miR-21	AI669815	SOX2	SRY (sex determining	P + T	establishment and/or maintenance of
			region Y)-box 2		chromatin
					architecture\|nucleus\|regulation of
					transcription, DNA-
					dependent\|transcription\|transcription
					factor activity
miR-21	NM_006940	SOX5	SRY (sex determining	P + T	nucleus\|regulation of transcription,
			region Y)-box 5		DNA-
					dependent\|transcription\|transcription
					factor activity\|transcription from
					RNA polymerase II promoter
miR-21	AI808807	SOX7	SRY (sex determining	P + T	DNA binding\|nucleus\|regulation of
			region Y)-box 7		transcription, DNA-
					dependent\|transcription
miR-21	NM_006717	SPIN	Spindling	P + T	gametogenesis\|ribonucleoprotein
					complex
miR-21	NM_005842	SPRY2	sprouty homolog 2	P + T	cell-cell
			(Drosophila)		signaling\|development\|mem-
					brane\|organogenesis\|regulation
					of signal transduction
miR-21	NM_006751	SSFA2	sperm specific antigen 2	P + T	plasma membrane
miR-21	NM_006603	STAG2	stromal antigen 2	P + T	cell cycle\|chromosome
					segregation\|cytokinesis\|meiosis\|mito-
					sis\|molecular_function
					unknown\|nucleus
miR-21	BC000627	STAT3	signal transducer and	P + T	JAK-STAT cascade\|acute-phase
			activator of transcription 3		response\|calcium ion binding\|cell
			(acute-phase response		motility\|cytoplasm\|hematopoietin\|interferon-
			factor)		class (D200-domain)
					cytokine receptor signal transducer
					activity\|intracellular signaling
					cascade\|negative regulation of
					transcription from RNA polymerase II
					promoter\|neurogenesis\|nucleus\|nu-
					cleus\|regulation of transcription, DNA-
					dependent\|signal transducer
					activity\|transcription\|transcription
					factor activity\|transcription factor
					activity
miR-21	AW138827	TAF5	TAF5 RNA polymerase II,	P + T	nucleus\|regulation of transcription,
			TATA box binding protein		DNA-dependent\|transcription factor
			(TBP)-associated factor,		TFIID complex\|transcription factor
			100 kDa		activity
miR-21	BF591040	TAGAP	T-cell activation GTPase	P + T	GTPase activator activity
			activating protein
miR-21	NM_000358	TGFBI	transforming growth factor,	M + P + T	cell adhesion\|cell
			beta-induced, 68 kDa		proliferation\|extracellular matrix
					(sensu Metazoa)\|extracellular
					space\|integrin binding\|negative
					regulation of cell adhesion\|protein
					binding\|sensory perception\|visual
					perception
miR-21	NM_000362	TIMP3	tissue inhibitor of	P + T	enzyme inhibitor
			metalloproteinase 3 (Sorsby		activity\|extracellular matrix (sensu
			fundus dystrophy,		Metazoa)\|extracellular matrix (sensu
			pseudoinflammatory)		Metazoa)\|induction of apoptosis by
					extracellular
					signals\|metalloendopeptidase
					inhibitor activity\|sensory
					perception\|visual perception
miR-21	AA149745	TRIM2	tripartite motif-containing 2	M + P + T	cytoplasm\|myosin binding\|protein
					ubiquitination\|ubiquitin ligase
					complex\|ubiquitin-protein ligase
					activity\|zinc ion binding
miR-21	AF346629	TRPM7	transient receptor potential	P + T	ATP binding\|calcium channel
			cation channel, subfamily		activity\|calcium ion transport\|cation
			M, member 7		transport\|integral to
					membrane\|membrane\|protein amino
					acid phosphorylation\|protein
					serine/threonine kinase
					activity\|transferase activity
miR-21	AI745185	YAP1	Yes-associated protein 1,	P + T
			65 kDa
miR-21	NM_005667	ZFP103	zinc finger protein 103	P + T	central nervous system
			homolog (mouse)		development\|integral to
					membrane\|protein
					ubiquitination\|ubiquitin ligase
					complex\|ubiquitin-protein ligase
					activity\|zinc ion binding
miR-21	N62196	ZNF367	zinc finger protein 367	M + P + T	nucleic acid binding\|nucleus\|zinc ion
					binding

M = MiRanda
P = PicTar
T = TargetScan

Example 3

Bio-Pathological Features and microRNA Expression

Materials and Methods
Immunohistochemical Analysis of Breast Cancer Samples.
Staining procedures were performed as described (Querzoli, P., et al., Anal. Quant. Cytol. Histol. 21:151-160 (1999)). Hormonal receptors were evaluated with 6F11 antibody for estrogen receptor α (ER) and PGR-1A6 antibody for progesterone receptor (PR) (Ventana, Tucson, Ariz., U.S.A.). The proliferation index was assessed with MIB1 antibody (DAKO, Copenhagen). ERBB2 was detected with CB11 antibody (Ventana, Tucson, Ariz., U.S.A.) and p53 protein expression was examined with DO7 antibody (Ventana, Tucson, Ariz., U.S.A.). Only tumor cells with distinct nuclear immunostaining for ER, PR, Mib1 and p53 were recorded as positive. Tumor cells were considered positive for ERBB2 when they showed distinct membrane immunoreactivity.
To perform a quantitative analysis of the expression of these various biological markers, the Eureka Menarini computerized image analysis system was used. For each tumor section, at least 20 microscopic fields of invasive carcinoma were measured using a 40× objective. The following cut-off values were employed: 10% of positive nuclear area for ER, PR, c-erbB2 and p53, 13% of nuclei expressing Mib1 was introduced to discriminate cases with high and low proliferative activity.
Results
To evaluate whether a correlation exists between various bio-pathological features associated with breast cancer and the expression of particular miRNAs, we generated and compared miRNA expression profiles for various cancer samples associated with the presence or absence of a particular breast cancer feature. In particular, we analyzed breast cancers with lobular or ductal histotypes, breast cancers with differential expression of either estrogen receptor alpha (ER) or progesterone receptor, and breast cancers with differences in lymph node metastasis, vascular invasion, proliferation index, and expression of ERBB2 and p53.
Expression profiles of lobular or ductal and +/−ERBB2 expression classes did not reveal any microRNAs that were differentially-expressed, while all other comparisons revealed a small number of differentially-expressed microRNAs (P<0.05). The results of this analysis are shown in FIG. 4.
Differentially-expressed miRNA families were identified for various bio-pathological features that are associated with human breast cancer. For example, all miR-30 miRNAs are down-regulated in both ER− and PR− tumors, suggesting that expression of miR-30 miRNAs is regulated by these hormones. In addition, the expression of various let-7 miRNAs was down-regulated in breast cancer samples with either lymph node metastasis or a high proliferation index, suggesting that reduced let-7 expression could be associated with a poor prognosis, a result that is consistent with previous findings. The discovery that the let-7 family of miRNAs regulates the expression of members of the RAS oncogene family provides a potential explanation for the role of let-7 miRNAs in human cancer.
miR-145 and miR-21, two miRNAs whose expression could differentiate cancer or normal tissues, were also differentially-expressed in cancers with a different proliferation index or different tumor stage. In particular, miR-145 is progressively down-regulated from normal breast to cancers with a high proliferation index. Similarly, miR-21 is progressively up-regulated from normal breast to cancers with high tumor stage. These findings suggest that deregulation of these two miRNAs may affect critical molecular events involved in tumor progression.
Another miRNA potentially involved in cancer progression is miR-9-3. miR-9-3 was downregulated in breast cancers with either high vascular invasion or lymph node metastasis, suggesting that its down-regulation was acquired during the course of tumor progression and, in particular, during the acquisition of metastatic potential.
The relevant teachings of all publications cited herein that have not explicitly been incorporated by reference, are incorporated herein by reference in their entirety. While this invention has been particularly shown and described with references to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims.

Claims

What is claimed is:

1. A pharmaceutical composition for treating breast cancer, comprising at least one isolated miR gene product, and a pharmaceutically-acceptable carrier;

wherein the at least one isolated miR gene product is selected from the group consisting of: miR-145, miR-10b, miR-123 (miR-126), miR-140-as, miR-125a, miR-125b-1, miR-125b-2, miR-194, miR-204, let-7a-2, let-7a-3, let-7d (let-7d-v1), let-7f-2, miR-101-1, miR-143 and combinations thereof.

2. A pharmaceutical composition for treating breast cancer, comprising at least one miR expression inhibitor compound, and a pharmaceutically-acceptable carrier:

wherein the at least one miR inhibitor compound is selected from the group consisting of miR-21, miR-155, miR-009-1 (miR-131-1), miR-34 (miR-170), miR-102 (miR-29b), miR-213, let-7i (let-7d-v2), miR-122a, miR-128b, miR-136, miR-149, miR-191, miR-196-1, miR-196-2, miR-202, miR-203, miR-206, miR-210, miR-213 and combinations thereof.

3. A pharmaceutical composition for treating breast cancer, comprising at least one isolated miR gene product, wherein the breast cancer is associated with one or more prognostic markers and the at least one miR gene product are selected from the group consisting of:

(i) the breast cancer is a breast cancer associated with estrogen receptor expression and the miR gene product is selected from the group consisting of miR-26a, miR-26b, miR-102 (miR-29b), miR-30a-5p, miR-30b, miR-30c, miR-30d, miR-185, miR-191, miR-206, miR-212, and combinations thereof;

(ii) the breast cancer is a breast cancer associated with progesterone receptor expression and the miR gene product is selected from the group consisting of let-7c, miR-26a, miR-29b, miR-30a-5p, miR-30b, miR-30c, miR-30d, and combinations thereof;

(iii) the breast cancer is a breast cancer associated with positive lymph node metastasis and the miR gene product is selected from the group consisting of let-7f-1, let-7a-3, let-7a-2, miR-9-3, and combinations thereof;

(iv) the breast cancer is a breast cancer associated with a high proliferative index and the miR gene product is selected from the group consisting of let-7c, let-7d, miR-26a, miR-26b, miR-30a-5p, miR-102, miR-145, and combinations thereof;

(v) the breast cancer is a breast cancer associated with detectable p53 expression and the miR gene product is selected from the group consisting of miR-16a, miR-128b and a combination thereof;

(vi) the breast cancer is a breast cancer associated with high vascular invasion and the miR gene product is selected from the group consisting of miR-9-3, miR-10b, miR-27a, miR-29a, miR-123, miR-205 and combinations thereof; and

(vii) the breast cancer is a breast cancer associated with an advanced tumor stage and the miR gene product is selected from the group consisting of miR-9-2, miR-15-a, miR-21, miR-30a-s, miR-133a-1, miR-137, miR-153-2, miR-154, miR-181a, miR-203, miR-213, and combinations thereof.