WO2015190542A1 - 胆道がんの検出キット又はデバイス及び検出方法 - Google Patents
胆道がんの検出キット又はデバイス及び検出方法 Download PDFInfo
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Definitions
- the present invention relates to a kit or device for detecting biliary tract cancer containing a nucleic acid capable of specifically binding to a specific miRNA, and a nucleic acid for use in testing for the presence or absence of biliary tract cancer in a subject.
- the present invention relates to a method for detecting biliary tract cancers, which comprises measuring the expression level of the miRNA using.
- the biliary tract refers to the entire excretion route until the bile secreted from hepatocytes flows into the duodenum, and is roughly divided into the intrahepatic bile duct in the liver and the extrahepatic biliary system outside the liver.
- the extrahepatic biliary system is the extrahepatic bile duct where bile is exported from the liver to the duodenum, the gallbladder that temporarily stores and concentrates bile, and the site where the bile duct and main pancreatic duct open to the duodenal lumen. It is roughly divided into three parts.
- biliary tract cancers are cancerous bile duct epithelial cells that cover the lumen, and chemotherapy and radiotherapy are less effective, and surgical resection by early detection is the only radical treatment.
- there is no subjective symptom in early biliary tract cancer for example, because the subjective symptoms such as jaundice and itching occur only when the cancer progresses and the bile duct is obstructed and the bile flows back into the blood vessels. Often done.
- intrahepatic bile duct cancer rarely obstructs extrahepatic bile ducts, many diseases progress without asymptomatic symptoms.
- extrahepatic bile duct cancer is the hepatic hilar region (hepatic hilar cholangiocarcinoma) at the entrance of the liver, the upper portion from the hilar portion to the gallbladder (upper bile duct cancer), and the middle portion from the gallbladder to the pancreas (central bile duct cancer) ),
- the lower part (lower bile duct cancer) from the pancreas to the duodenal papilla, and the cancer that has occurred in the bile duct near the liver is known to be difficult to operate and the prognosis is poor.
- the degree of progression of extrahepatic bile duct cancer, gallbladder cancer and papillary cancer by UICC is “Biliary Tract Cancer Handling Code 5th Edition” (Edited by Japan Biliary Surgery Study Group, Kanbara Publishing Co., Ltd., 2003). P109) and classified into stages 0, IA, IB, IIA, IIB, III, IVa, and IVb according to lymph node metastasis, extra-abdominal distant other organ metastasis, macroscopic bile duct progression, and the like.
- the progression of intrahepatic cholangiocarcinoma by UICC is defined in “TNM Malignant Tumor Classification, 7th Edition, Japanese Version” (Translated by UICC Japan Committee, TNM Committee, Kanbara Publishing Co., Ltd., 2012, p110) It is classified into stages I, II, III, IVa, and IVb according to lymph node metastasis, extra-abdominal distant other organ metastasis, macroscopic bile duct progress, and the like.
- Non-patent Document 1 For the first diagnosis of biliary tract cancer, generally, a minimally invasive blood biochemical test, a tumor marker test, and an abdominal ultrasound test are used (Non-patent Document 1).
- blood biochemical tests for detecting biliary tract cancer for example, alkaline phosphatase, ⁇ -GTP, bilirubin and the like that increase due to impaired liver function are used.
- Known tumor markers for detecting biliary tract cancer include, for example, CEA, CA19-9, DUPAN-2, CA195, CA242, IL-6, and the like.
- cancer is suspected when the blood concentration is higher or lower than a predetermined reference value.
- the CEA reference value is set to 5 ng / mL
- the CA19-9 reference value is set to 37 U / mL. Cancer including cancer is suspected.
- Patent Document 1 describes a method for detecting biliary tract cancer using the expression level of protein in biliary tissue.
- Patent Document 2 describes a method for diagnosing digestive organ cancer including biliary tract cancer using mRNA gene extracted from cells (mononuclear cells, etc.) in blood.
- Biliary tract cancer clinical practice guideline publishing committee compilation "biliary tract cancer clinical practice guideline based on evidence", medical book publication corporation, 2007, p38-39 Kurokawa Kiyoshi, Clinical Laboratory Data Book, 2013, p633, 636
- An object of the present invention is to find a novel biliary tract cancer tumor marker and provide a method capable of effectively detecting biliary tract cancer using a nucleic acid that can specifically bind to the marker.
- a minimally invasive blood biochemical test, tumor marker test, and abdominal ultrasonography are generally used for the initial diagnosis of biliary tract cancer.
- Tumor visualization rate of biliary tract cancer by abdominal ultrasonography varies from 21 to 90% (Non-patent Document 1), especially when the cancer occupied site is the lower bile duct The rate will drop.
- In blood biochemical tests for example, alkaline phosphatase, ⁇ -GTP, bilirubin, etc.
- Non-patent Document 1 describes that it is not specific to cancer and difficult to use for early diagnosis.
- Non-patent document 1 describes that the clinical utility of DUPAN-2, CA195, CA242, and IL-6 is not clear. Therefore, when a conventional tumor marker is used, there is a possibility of misdetecting other cancers and / or benign tumors and / or benign diseases of the biliary tract and / or organs around the biliary tract.
- Patent Document 1 describes a method for detecting biliary tract cancer using the expression level of protein in biliary tissue.
- tissue excision by surgery is indispensable for obtaining a specimen, and this process is not preferable as an inspection method because the physical burden imposed on the patient is heavy.
- Patent Document 1 does not describe the detection performance such as specific accuracy, sensitivity, specificity, etc. for discriminating biliary tract cancers with respect to this detection method, and is poor in industrial practicality.
- Patent Document 2 describes a method for diagnosing digestive organ cancer including biliary tract cancer using mRNA gene extracted from cells (mononuclear cells, etc.) in blood.
- this detection method needs to use a combination of several tens to several hundreds of mRNAs, and when actually developed as a test, there are concerns about an increase in test cost and complexity of the discrimination algorithm.
- mRNA is easily decomposed in blood and unstable, it is not preferable as a test target.
- the present inventors have found several genes that can be used as a marker for detection of biliary tract cancer from blood that can be collected in a minimally invasive manner, and have developed a nucleic acid that can specifically bind thereto. By using it, it was found that biliary tract cancer can be detected significantly, and the present invention has been completed.
- Biliary tract cancer markers miR-125a-3p, miR-6893-5p, miR-204-3p, miR-4476, miR-4294, miR-150-3p, miR-6729-5p, miR- 7641, miR-6765-3p, miR-6820-5p, miR-575, miR-6683-3p, miR-1469, miR-663a, miR-6075, miR-4634, miR-423-5p, miR-4454, miR-7109-5p, miR-6789-5p, miR-6877-5p, miR-4789, miR-4530, miR-7975, miR-6724-5p, miR-8073, miR-7777, miR-1231, miR- 6799-5p, miR-615-5p, miR-4450 miR-6726-5p, miR-6875-5p, miR-4734, miR-16
- miR-125a-3p is hsa-miR-125a-3p
- miR-6893-5p is hsa-miR-6893-5p
- miR-204-3p is hsa-miR-204-3p
- MiR-4476 is hsa-miR-4476
- miR-4294 is hsa-miR-4294
- miR-150-3p is hsa-miR-150-3p
- miR-6729-5p is hsa-miR.
- miR-7641 is hsa-miR-7641
- miR-6765-3p is hsa-miR-6765-3p
- miR-6820-5p is hsa-miR-6820-5p
- MiR-575 is hsa-miR-575
- miR-6683-3p is hsa-miR 6836-3p
- miR-1469 is hsa-miR-1469
- miR-663a is hsa-miR-663a
- miR-6075 is hsa-miR-6075
- miR-4634 is hsa-miR- 4634
- miR-423-5p is hsa-miR-423-5p
- miR-4454 is hsa-miR-4454
- miR-7109-5p is hsa-miR-7109-5p
- miR- 6789-5p is hsa-miR
- the kit is another biliary tract cancer marker, miR-6808-5p, miR-6774-5p, miR-4656, miR-6806-5p, miR-1233-5p, miR-328-5p, miR-4675, miR-2110, miR-6076, miR-3619-3p, miR-92a-2-5p, miR-128-1-5p, miR-638, miR-2861, miR-371a-5p, miR- At least one poly selected from the group consisting of 211-3p, miR-1273g-3p, miR-1203, miR-122-5p, miR-4258, miR-4484, miR-4648 and miR-6780b-5p
- the nucleic acid according to any one of (1) to (3), further comprising a nucleic acid capable of specifically binding to a nucleotide. Kit.
- miR-6808-5p is hsa-miR-6808-5p
- miR-6774-5p is hsa-miR-6774-5p
- miR-4656 is hsa-miR-4656
- miR-6806 -5p is hsa-miR-6806-5p
- miR-1233-5p is hsa-miR-1233-5p
- miR-328-5p is hsa-miR-328-5p
- miR-4673 is hsa MiR-4675
- miR-2110 is hsa-miR-2110
- miR-6076 is hsa-miR-6076
- miR-3619-3p is hsa-miR-3619-3p
- miR-92a -2-5p is hsa-miR-92a-2-5p
- miR-128-1-5p is sa-miR-128-1-5p
- the kit comprises at least two or more nucleic acids capable of specifically binding to each of at least two or more polynucleotides selected from all the biliary tract cancer markers described in (1) or (2).
- Biliary tract cancer markers miR-125a-3p, miR-6893-5p, miR-204-3p, miR-4476, miR-4294, miR-150-3p, miR-6729-5p, miR- 7641, miR-6765-3p, miR-6820-5p, miR-575, miR-6683-3p, miR-1469, miR-663a, miR-6075, miR-4634, miR-423-5p, miR-4454, miR-7109-5p, miR-6789-5p, miR-6877-5p, miR-4789, miR-4530, miR-7975, miR-6724-5p, miR-8073, miR-7777, miR-1231, miR- 6799-5p, miR-615-5p, miR-4450 miR-6726-5p, miR-6875-5p, miR-4734, miR-16-5p, miR-602, miR-4651, miR-8069,
- miR-125a-3p is hsa-miR-125a-3p
- miR-6893-5p is hsa-miR-6893-5p
- miR-204-3p is hsa-miR-204-3p
- MiR-4476 is hsa-miR-4476
- miR-4294 is hsa-miR-4294
- miR-150-3p is hsa-miR-150-3p
- miR-6729-5p is hsa-miR.
- miR-7641 is hsa-miR-7641
- miR-6765-3p is hsa-miR-6765-3p
- miR-6820-5p is hsa-miR-6820-5p
- MiR-575 is hsa-miR-575
- miR-6683-3p is hsa-miR 6836-3p
- miR-1469 is hsa-miR-1469
- miR-663a is hsa-miR-663a
- miR-6075 is hsa-miR-6075
- miR-4634 is hsa-miR- 4634
- miR-423-5p is hsa-miR-423-5p
- miR-4454 is hsa-miR-4454
- miR-7109-5p is hsa-miR-7109-5p
- miR- 6789-5p is hsa-miR
- the device is another biliary tract cancer marker, miR-6808-5p, miR-6774-5p, miR-4656, miR-6806-5p, miR-1233-5p, miR-328-5p, miR-4675, miR-2110, miR-6076, miR-3619-3p, miR-92a-2-5p, miR-128-1-5p, miR-638, miR-2861, miR-371a-5p, miR- At least one poly selected from the group consisting of 211-3p, miR-1273g-3p, miR-1203, miR-122-5p, miR-4258, miR-4484, miR-4648 and miR-6780b-5p Any of (8) to (10), further comprising a nucleic acid capable of specifically binding to a nucleotide The device according to.
- miR-6808-5p is hsa-miR-6808-5p
- miR-6774-5p is hsa-miR-6774-5p
- miR-4656 is hsa-miR-4656
- miR-6806 -5p is hsa-miR-6806-5p
- miR-1233-5p is hsa-miR-1233-5p
- miR-328-5p is hsa-miR-328-5p
- miR-4673 is hsa MiR-4675
- miR-2110 is hsa-miR-2110
- miR-6076 is hsa-miR-6076
- miR-3619-3p is hsa-miR-3619-3p
- miR-92a -2-5p is hsa-miR-92a-2-5p
- miR-128-1-5p hsa-miR-128-1-5p
- the hybridization technique is a nucleic acid array technique.
- the device comprises at least two or more nucleic acids capable of specifically binding to each of at least two or more polynucleotides selected from all the biliary tract cancer markers according to (8) or (9).
- a method for detecting biliary tract cancer comprising evaluating by (18) The method according to (17), wherein the subject is a human. (19) The method according to (17) or (18), wherein the specimen is blood, serum or plasma.
- biliary tract cancer refers to all malignant tumors formed in the biliary tract. Specifically, extrahepatic bile duct cancer, gallbladder cancer, papillary cancer, duodenal papillary cancer, intrahepatic bile duct cancer and the like are included.
- the “benign tumor and / or benign disease of the biliary tract and / or peripheral organs of the biliary tract” refers to a disease of a non-malignant tumor related to the biliary tract, liver, and pancreas.
- polynucleotide is used for nucleic acids including RNA, DNA, and RNA / DNA (chimera).
- the DNA includes any of cDNA, genomic DNA, and synthetic DNA.
- the RNA includes total RNA, mRNA, rRNA, miRNA, siRNA, snoRNA, snRNA, non-coding RNA and synthetic RNA.
- synthetic DNA and “synthetic RNA” are artificially generated using, for example, an automatic nucleic acid synthesizer based on a predetermined base sequence (which may be either a natural sequence or a non-natural sequence). It refers to the prepared DNA and RNA.
- non-natural sequence is intended to be used in a broad sense, and is a sequence (for example, one or more nucleotide substitutions, deletions, insertions and / or additions) that differs from the natural sequence ( That is, it includes a mutant sequence), a sequence containing one or more modified nucleotides (ie, a modified sequence), and the like.
- the polynucleotide is used interchangeably with the nucleic acid.
- a “fragment” is a polynucleotide having a base sequence of a continuous part of a polynucleotide, and desirably has a length of 15 bases or more, preferably 17 bases or more, more preferably 19 bases or more. .
- RNA and double-stranded DNA include not only RNA and double-stranded DNA, but also each single-stranded DNA such as positive strand (or sense strand) or complementary strand (or antisense strand) constituting the same. It is intended to be used.
- the length is not particularly limited.
- “gene” refers to double-stranded DNA containing human genomic DNA, single-stranded DNA containing cDNA (positive strand), and single-stranded DNA having a sequence complementary to the positive strand.
- DNA complementary strand
- miRNA microRNA
- transcripts fragments thereof, and transcripts are all included.
- the “gene” is not limited to a “gene” represented by a specific nucleotide sequence (or sequence number), but also RNAs having biological functions equivalent to RNA encoded by these, for example, homologs (ie, homologs). Or orthologs), variants such as genetic polymorphisms, and “nucleic acids” encoding derivatives.
- the “nucleic acid” encoding such homologue, variant or derivative is, for example, the base sequence represented by any one of SEQ ID NOs: 1 to 509 under the stringent conditions described later, or the base A “nucleic acid” having a base sequence that hybridizes with a complementary sequence of the base sequence in which u is t in the sequence can be mentioned.
- the “gene” does not ask whether the functional region is different, and may include, for example, an expression control region, a coding region, an exon, or an intron. Further, the “gene” may be contained in the cell, may be released outside the cell and may be present alone, or may be encapsulated in a vesicle called an exosome.
- exosome is a vesicle encased in a lipid bilayer secreted from a cell. Exosomes are derived from multivesicular endosomes, and when released to the extracellular environment, they may contain biological substances such as “genes” such as RNA and DNA and proteins. It is known that exosomes are contained in body fluids such as blood, serum, plasma, serum and lymph.
- RNA refers to RNA synthesized using a DNA sequence of a gene as a template.
- RNA is synthesized in such a manner that RNA polymerase binds to a site called a promoter located upstream of the gene and ribonucleotides are bound to the 3 'end so as to be complementary to the DNA base sequence.
- This RNA includes not only the gene itself but also the entire sequence from the transcription start point to the end of the poly A sequence, including the expression control region, coding region, exon or intron.
- microRNA is a protein complex that is transcribed as a hairpin-like RNA precursor, cleaved by a dsRNA cleaving enzyme having RNase III cleaving activity, and called RISC. 15-25 base non-coding RNA that is incorporated into and is involved in the translational repression of mRNA.
- miRNA is not limited to “miRNA” represented by a specific nucleotide sequence (or sequence number), but also a precursor of the “miRNA” (pre-miRNA, pri-miRNA), and these And miRNAs that have equivalent biological functions, such as homologs (ie, homologs or orthologs), variants such as genetic polymorphisms, and derivatives. Such precursors, homologues, mutants or derivatives can be specifically identified by miRBase release 20 (http://www.mirbase.org/), and under stringent conditions described later. And “miRNA” having a base sequence that hybridizes with a complementary sequence of any one of the specific base sequences represented by any of SEQ ID NOs: 1 to 509.
- miRNA used herein may be a gene product of a miR gene, and such a gene product is a mature miRNA (for example, 15 to 15 involved in the suppression of translation of mRNA as described above). 25-base, or 19-25 base non-coding RNA) or miRNA precursors (eg, pre-miRNA or pri-miRNA as described above).
- the “probe” includes a polynucleotide used for specifically detecting RNA produced by gene expression or a polynucleotide derived therefrom and / or a polynucleotide complementary thereto.
- the “primer” includes a polynucleotide that specifically recognizes and amplifies RNA generated by gene expression or a polynucleotide derived therefrom and / or a polynucleotide complementary thereto.
- a complementary polynucleotide is a polynucleotide comprising a base sequence defined by any of SEQ ID NOs: 1 to 509 or a base sequence in which u is t in the base sequence.
- the base sequence is complementary to the full-length sequence or a partial sequence thereof (for convenience, this is referred to as the positive strand) based on the base pair relationship such as A: T (U), G: C.
- such a complementary strand is not limited to the case where it forms a completely complementary sequence with the target positive strand base sequence, but has a complementary relationship that allows hybridization with the target normal strand under stringent conditions. There may be.
- stringent conditions refers to the degree to which a nucleic acid probe is larger than other sequences (for example, the average of background measurement values + standard error of background measurement values ⁇ 2 or more measurement values) The conditions for hybridizing to the target sequence. Stringent conditions are sequence-dependent and depend on the environment in which hybridization is performed. By controlling the stringency of the hybridization and / or wash conditions, target sequences that are 100% complementary to the nucleic acid probe can be identified. Specific examples of “stringent conditions” will be described later.
- the “Tm value” means a temperature at which the double-stranded portion of the polynucleotide is denatured into a single strand and the double strand and the single strand are present at a ratio of 1: 1.
- variant refers to a natural variant caused by polymorphism, mutation, etc., or any nucleotide sequence of SEQ ID NOs: 1 to 509, or u in the nucleotide sequence in the case of nucleic acid.
- nucleotide sequence that is t, or a variant that includes deletion, substitution, addition, or insertion of one or more bases in the partial sequence, or each of the base sequence or a partial sequence thereof, and about 90% or more, about 95% Above, about 97% or more, about 98% or more, about 99% or more of a variant exhibiting% identity, or a polynucleotide or oligonucleotide containing the base sequence or a partial sequence thereof under the stringent conditions defined above. It means nucleic acid that soy.
- “several” means an integer of about 10, 9, 8, 7, 6, 5, 4, 3 or 2.
- a mutant can be prepared using a well-known technique such as site-directed mutagenesis or PCR-based mutagenesis.
- % identity can be determined using the above-described BLAST or FASTA protein or gene search system with or without introducing a gap (Zheng Zhang et al., 2000, J. Comput. Biol., 7, p203-214; Altschul, SF, et al., 1990, Journal of Molecular Biology, 215, p403-410; Pearson, WR, et al., 1988. (Proc. Natl. Acad. Sci. USA, vol. 85, p2444-448).
- the term “derivative” refers to a modified nucleic acid, a non-limiting group such as a labeled derivative such as a fluorophore, a modified nucleotide (for example, a halogen, an alkyl such as methyl, an alkoxy such as methoxy, a group such as thio, carboxymethyl, etc.
- a derivative containing PNA peptide nucleic acid; Nielsen, PE, etc.). 1991, Science, 254, p1497-500
- LNA locked nucleic acid; Obika, S. et al., 1998, Tetrahedron Lett., 39, p5401-5404) and the like.
- nucleic acid that can specifically bind to a polynucleotide selected from the group of miRNAs that are the above-mentioned biliary tract cancer markers is a nucleic acid that is synthesized or prepared.
- nucleotides, oligonucleotides and polynucleotides capable of binding are used as probes for detecting the gene expressed in vivo, in tissues or cells based on the above properties, and for amplifying the gene expressed in vivo. It can be effectively used as a primer.
- the term “detection” can be replaced by the term inspection, measurement, detection or decision support. Further, in this specification, the term “evaluation” is used in a meaning including supporting diagnosis or evaluation based on a test result or a measurement result.
- subject includes humans, primates including chimpanzees, pet animals such as dogs and cats, livestock animals such as cows, horses, sheep and goats, rodents such as mice and rats, etc. Means a mammal.
- healthy body also means an animal that is such a mammal and does not suffer from the cancer to be detected.
- P or “P value” refers to the probability that, in a statistical test, a statistic more extreme than the statistic actually calculated from the data under the null hypothesis is observed. Indicates. Therefore, it can be considered that the smaller the “P” or “P value”, the more significant the difference between the comparison objects.
- sensitivity means a value of (number of true positives) / (number of true positives + number of false negatives). High sensitivity makes it possible to detect biliary tract cancer at an early stage, leading to complete removal of the cancerous part and a reduction in the recurrence rate.
- specificity means (number of true negatives) / (number of true negatives + number of false positives). If the specificity is high, it is possible to prevent unnecessary additional examinations by misidentifying a healthy body as a patient with biliary tract cancer, thereby reducing the burden on the patient and reducing medical costs.
- accuracy means a value of (number of true positives + number of true negatives) / (number of all cases). The accuracy indicates the rate at which the discrimination results for all the samples are correct, and is a first index for evaluating the detection performance.
- the “specimen” to be determined, detected or diagnosed changes the expression of the gene of the present invention as the biliary tract cancer develops, the biliary tract cancer progresses, and the therapeutic effect on the biliary tract cancer is exerted.
- tissue and biomaterial Specifically, biliary tissues and their surrounding vessels, lymph nodes and organs, organs suspected of metastasis, skin, and body fluids such as blood, urine, saliva, sweat, and tissue exudates, and serum and plasma prepared from blood Others include stool and hair. Furthermore, it refers to a biological sample extracted from these, specifically genes such as RNA and miRNA.
- hsa-miR-125a-3p gene or “hsa-miR-125a-3p” refers to the hsa-miR-125a-3p gene described in SEQ ID NO: 1 (miRBase Accession No. 1). MIMAT0004602) and other species homologs or orthologs.
- the hsa-miR-125a-3p gene can be obtained by the method described in Lagos-Quintana M et al., 2002, Curr Biol, 12, p735-739.
- “hsa-miR-125a-3p” “hsa-mir-125a” (miRBase Accession No. MI000069, SEQ ID NO: 149) having a hairpin-like structure as a precursor is known.
- hsa-miR-6893-5p gene or “hsa-miR-6893-5p” refers to the hsa-miR-6893-5p gene described in SEQ ID NO: 2 (miRBase Accession No. MIMAT0027686) and other species homologs or orthologs.
- the hsa-miR-6893-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6893-5p “hsa-mir-6893” (miRBase Accession No. MI0022740, SEQ ID NO: 150) having a hairpin-like structure as a precursor is known.
- hsa-miR-204-3p gene or “hsa-miR-204-3p” refers to the hsa-miR-204-3p gene (miRBase Accession No. 3) described in SEQ ID NO: 3. MIMAT0022693) and other species homologs or orthologs.
- the hsa-miR-204-3p gene can be obtained by the method described in Lim LP et al., 2003, Science, 299, p1540.
- “hsa-miR-204-3p” is known as “hsa-mir-204” (miRBase Accession No. MI00000028, SEQ ID NO: 151) having a hairpin-like structure as a precursor.
- hsa-miR-4476 gene or “hsa-miR-4476” refers to the hsa-miR-4476 gene (miRBase Accession No. MIMAT0019003) described in SEQ ID NO: 4 and other biological species. Includes homologs or orthologs.
- the hsa-miR-4476 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-miR-4476 is known as “hsa-mir-4476” (miRBase Accession No. MI0016828, SEQ ID NO: 152) having a hairpin-like structure as a precursor.
- hsa-miR-4294 gene or “hsa-miR-4294” refers to the hsa-miR-4294 gene (miRBase Accession No. MIMAT0016849) described in SEQ ID NO: 5 and other biological species. Includes homologs or orthologs.
- the hsa-miR-4294 gene can be obtained by the method described in Goff LA et al., 2009, PLoS One, vol. 4, e7192.
- hsa-miR-4294 “hsa-mir-4294” (miRBase Accession No. MI0015827, SEQ ID NO: 153) having a hairpin-like structure as a precursor is known.
- hsa-miR-150-3p gene or “hsa-miR-150-3p” refers to the hsa-miR-150-3p gene described in SEQ ID NO: 6 (miRBase Accession No. MIMAT0004610) and other species homologs or orthologs.
- the hsa-miR-150-3p gene can be obtained by the method described in Lagos-Quintana M et al., 2002, Curr Biol, 12, p735-739.
- “hsa-miR-150-3p” is known as “hsa-mir-150” (miRBase Accession No. MI000079, SEQ ID NO: 154) having a hairpin-like structure as a precursor.
- hsa-miR-6729-5p gene or “hsa-miR-6729-5p” refers to the hsa-miR-6729-5p gene (miRBase Accession No. 7) described in SEQ ID NO: 7. MIMAT0027359) and other species homologs or orthologs.
- the hsa-miR-6729-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6729-5p “hsa-mir-6729” (miRBase Accession No. MI0022574, SEQ ID NO: 155) having a hairpin-like structure as a precursor is known.
- hsa-miR-7641 gene or “hsa-miR-7641” refers to the hsa-miR-7641 gene (miRBase Accession No. MIMAT0029782) described in SEQ ID NO: 8 or other biological species. Includes homologs or orthologs.
- the hsa-miR-7641 gene can be obtained by the method described in Yo JK et al., 2013, Arch PharmRes, 36, p353-358.
- “Hsa-miR-7641” has a hairpin-like structure as a precursor thereof, “hsa-mir-7464-1”, “hsa-mir-7641-2” (miRBase Accession No. MI0024975, MI0024976, SEQ ID NO: 156, 157) are known.
- hsa-miR-6765-3p gene or “hsa-miR-6765-3p” refers to the hsa-miR-6765-3p gene described in SEQ ID NO: 9 (miRBase Accession No. MIMAT0027431) and other species homologs or orthologs.
- the hsa-miR-6765-3p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6765-3p “hsa-mir-6765” (miRBase Accession No. MI0022610, SEQ ID NO: 158) having a hairpin-like structure as a precursor is known.
- hsa-miR-6820-5p gene or “hsa-miR-6820-5p” refers to the hsa-miR-6820-5p gene (miRBase Accession No. 5) described in SEQ ID NO: 10. MIMAT0027540) and other species homologs or orthologs.
- the hsa-miR-6820-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p1634-1645.
- hsa-miR-6820-5p “hsa-mir-6820” (miRBase Accession No. MI0022665, SEQ ID NO: 159) having a hairpin-like structure as a precursor is known.
- hsa-miR-575 gene or “hsa-miR-575” refers to the hsa-miR-575 gene (miRBase Accession No. MIMAT0003240) described in SEQ ID NO: 11 and other biological species. Includes homologs or orthologs.
- the hsa-miR-575 gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci U S A, 103, p3687-3692.
- hsa-miR-575 “hsa-mir-575” (miRBase Accession No. MI0003582, SEQ ID NO: 160) having a hairpin-like structure as a precursor is known.
- hsa-miR-6836-3p gene or “hsa-miR-6836-3p” refers to the hsa-miR-6636-3p gene (miRBase Accession No. 1) described in SEQ ID NO: 12. MIMAT0027575) and other species homologs or orthologs.
- the hsa-miR-68336p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-6836-3p” is known as “hsa-mir-6683” (miRBase Accession No. MI0022682, SEQ ID NO: 161), which has a hairpin-like structure as a precursor.
- hsa-miR-1469 gene or “hsa-miR-1469” refers to the hsa-miR-1469 gene (miRBase Accession No. MIMAT0007347) described in SEQ ID NO: 13 and other biological species. Includes homologs or orthologs.
- the hsa-miR-1469 gene can be obtained by the method described in Kawaji H et al., 2008, BMC Genomics, Vol. 9, p157.
- hsa-miR-1469 “hsa-mir-1469” (miRBase Accession No. MI00000074, SEQ ID NO: 162) having a hairpin-like structure as a precursor is known.
- hsa-miR-663a gene or “hsa-miR-663a” refers to the hsa-miR-663a gene (miRBase Accession No. MIMAT0003326) described in SEQ ID NO: 14 or other biological species. Includes homologs or orthologs.
- the hsa-miR-663a gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci U S A, 103, p3687-3692.
- “hsa-miR-663a” is known as “hsa-mir-663a” (miRBase Accession No. MI0003672, SEQ ID NO: 163) having a hairpin-like structure as a precursor.
- hsa-miR-6075 gene or “hsa-miR-6075” refers to the hsa-miR-6075 gene (miRBase Accession No. MIMAT0023700) described in SEQ ID NO: 15 and other biological species. Includes homologs or orthologs.
- the hsa-miR-6075 gene can be obtained by the method described in Voellenkle C et al., 2012, RNA, 18, p472-484.
- hsa-miR-6075 is known as “hsa-mir-6075” (miRBase Accession No. MI0020352, SEQ ID NO: 164) having a hairpin-like structure as a precursor.
- hsa-miR-4634 gene or “hsa-miR-4634” refers to the hsa-miR-4634 gene (miRBase Accession No. MIMAT0019691) described in SEQ ID NO: 16 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4634 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, 71, p78-86.
- “hsa-miR-4634” is known as “hsa-mir-4634” (miRBase Accession No. MI0017261, SEQ ID NO: 165) having a hairpin-like structure as a precursor.
- hsa-miR-423-5p gene or “hsa-miR-423-5p” refers to the hsa-miR-423-5p gene described in SEQ ID NO: 17 (miRBase Accession No. MIMAT0004748) and other species homologs or orthologs.
- the hsa-miR-423-5p gene can be obtained by the method described in Kasashima K et al., 2004, Biochem Biophys Res Commun, 322, p403-410.
- hsa-miR-423-5p “hsa-mir-423” (miRBase Accession No. MI0001445, SEQ ID NO: 166) having a hairpin-like structure as a precursor is known.
- hsa-miR-4454 gene or “hsa-miR-4454” refers to the hsa-miR-4454 gene (miRBase Accession No. MIMAT0018976) described in SEQ ID NO: 18 and other biological species. Includes homologs or orthologs.
- the hsa-miR-4454 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-mir-4454 (miRBase Accession No. MI0016800, SEQ ID NO: 167) having a hairpin-like structure as a precursor is known.
- hsa-miR-7109-5p gene or “hsa-miR-7109-5p” refers to the hsa-miR-7109-5p gene described in SEQ ID NO: 19 (miRBase Accession No. MIMAT0028115) and other species homologs or orthologs.
- the hsa-miR-7109-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-7109-5p” is known as “hsa-mir-7109” (miRBase Accession No. MI0022960, SEQ ID NO: 168), which has a hairpin-like structure as a precursor.
- hsa-miR-6789-5p gene or “hsa-miR-6789-5p” refers to the hsa-miR-6789-5p gene described in SEQ ID NO: 20 (miRBase Accession No. MIMAT0027478) and other species homologs or orthologs.
- the hsa-miR-6789-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6789-5p “hsa-mir-6789” (miRBase Accession No. MI0022634, SEQ ID NO: 169) having a hairpin-like structure as a precursor is known.
- hsa-miR-6877-5p gene or “hsa-miR-6877-5p” refers to the hsa-miR-6877-5p gene described in SEQ ID NO: 21 (miRBase Accession No. MIMAT0027654) and other species homologs or orthologs.
- the hsa-miR-6877-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-6877-5p” is known as “hsa-mir-6877” (miRBase Accession No. MI0022724, SEQ ID NO: 170) having a hairpin-like structure as a precursor.
- hsa-miR-4792 gene or “hsa-miR-4792” refers to the hsa-miR-4792 gene (miRBase Accession No. MIMAT0019964) described in SEQ ID NO: 22 and other biological species. Includes homologs or orthologs.
- the hsa-miR-4792 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, 71, p78-86.
- hsa-miR-4792 “hsa-mir-4792” (miRBase Accession No. MI0017439, SEQ ID NO: 171) having a hairpin-like structure as a precursor is known.
- hsa-miR-4530 gene or “hsa-miR-4530” refers to the hsa-miR-4530 gene (miRBase Accession No. MIMAT0019069) described in SEQ ID NO: 23 and other species. Includes homologs or orthologs.
- the hsa-miR-4530 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-miR-4530 is known as “hsa-mir-4530” (miRBase Accession No. MI0016897, SEQ ID NO: 172), which has a hairpin-like structure as a precursor.
- hsa-miR-7975 gene or “hsa-miR-7975” refers to the hsa-miR-7975 gene (miRBase Accession No. MIMAT0031178) described in SEQ ID NO: 24 and other biological species. Includes homologs or orthologs.
- the hsa-miR-7975 gene can be obtained by the method described in Velthut-Meikas A et al., 2013, Mol Endocrinol, online edition.
- “hsa-miR-7975” is known as “hsa-mir-7975” (miRBase Accession No. MI0025751, SEQ ID NO: 173) having a hairpin-like structure as a precursor.
- hsa-miR-6724-5p gene or “hsa-miR-6724-5p” refers to the hsa-miR-6724-5p gene described in SEQ ID NO: 25 (miRBase Accession No. MIMAT0025856) and other species homologs or orthologs.
- the hsa-miR-6724-5p gene can be obtained by the method described in Li Y et al., 2012, Gene, 497, p330-335.
- “hsa-miR-6724-5p” “hsa-mir-6724” (miRBase Accession No. MI0022559, SEQ ID NO: 174) having a hairpin-like structure as a precursor is known.
- hsa-miR-8073 gene or “hsa-miR-8073” refers to the hsa-miR-8073 gene (miRBase Accession No. MIMAT0031000) described in SEQ ID NO: 26 or other biological species. Includes homologs or orthologs.
- the hsa-miR-8073 gene can be obtained by the method described in Wang HJ et al., 2013, Shock, 39, p480-487.
- hsa-miR-8073 “hsa-mir-8073” (miRBase Accession No. MI0025909, SEQ ID NO: 175) having a hairpin-like structure as a precursor is known.
- hsa-miR-7777 gene or “hsa-miR-7777” refers to the hsa-miR-7777 gene (miRBase Accession No. MIMAT0031180) described in SEQ ID NO: 27 and other biological species. Includes homologs or orthologs.
- the hsa-miR-7777 gene can be obtained by the method described in Velthut-Meikas A et al., 2013, Mol Endocrinol, online edition.
- hsa-miR-7777 is known as “hsa-mir-7777” (miRBase Accession No. MI0025753, SEQ ID NO: 176) having a hairpin-like structure as a precursor.
- hsa-miR-1231 gene or “hsa-miR-1231” refers to the hsa-miR-1231 gene (miRBase Accession No. MIMAT0005586) described in SEQ ID NO: 28 or other biological species. Includes homologs or orthologs.
- the hsa-miR-1231 gene can be obtained by the method described in Berezkov E et al., 2007, Mol Cell, 28, p328-336.
- hsa-mir-1231 miRBase Accession No. MI0006321, SEQ ID NO: 177) having a hairpin-like structure as a precursor is known.
- hsa-miR-6799-5p gene or “hsa-miR-6799-5p” refers to the hsa-miR-6799-5p gene described in SEQ ID NO: 29 (miRBase Accession No. MIMAT0027498) and other species homologs or orthologs.
- the hsa-miR-6799-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6799-5p “hsa-mir-6799” (miRBase Accession No. MI0022644, SEQ ID NO: 178) having a hairpin-like structure as a precursor is known.
- hsa-miR-615-5p gene or “hsa-miR-615-5p” refers to the hsa-miR-615-5p gene described in SEQ ID NO: 30 (miRBase Accession No. MIMAT0004804) and other species homologs or orthologs.
- the hsa-miR-615-5p gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci U S A, 103, p3687-3692.
- “Hsa-miR-615-5p” is known as “hsa-mir-615” (miRBase Accession No. MI0003628, SEQ ID NO: 179) having a hairpin-like structure as a precursor.
- hsa-miR-4450 gene or “hsa-miR-4450” refers to the hsa-miR-4450 gene (miRBase Accession No. MIMAT0018971) described in SEQ ID NO: 31 and other species. Includes homologs or orthologs.
- the hsa-miR-4450 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-mir-4450 (miRBase Accession No. MI0016795, SEQ ID NO: 180) having a hairpin-like structure as a precursor is known.
- hsa-miR-6726-5p gene or “hsa-miR-6726-5p” refers to the hsa-miR-6726-5p gene described in SEQ ID NO: 32 (miRBase Accession No. MIMAT0027353) and other species homologs or orthologs.
- the hsa-miR-6726-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-6726-5p” is known as “hsa-mir-6726” (miRBase Accession No. MI0022571, SEQ ID NO: 181) having a hairpin-like structure as a precursor.
- hsa-miR-6875-5p gene or “hsa-miR-6875-5p” refers to the hsa-miR-6875-5p gene described in SEQ ID NO: 33 (miRBase Accession No. MIMAT0027650) and other species homologs or orthologs.
- the hsa-miR-6875-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6875-5p “hsa-mir-6875” (miRBase Accession No. MI0022722, SEQ ID NO: 182) having a hairpin-like structure as a precursor is known.
- hsa-miR-4734 gene or “hsa-miR-4734” refers to the hsa-miR-4734 gene (miRBase Accession No. MIMAT0019859) described in SEQ ID NO: 34 and other biological species. Includes homologs or orthologs.
- the hsa-miR-4734 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- “Hsa-miR-4734” is known as “hsa-mir-4734” (miRBase Accession No. MI0017371, SEQ ID NO: 183), which has a hairpin-like structure as a precursor.
- hsa-miR-16-5p gene or “hsa-miR-16-5p” refers to the hsa-miR-16-5p gene described in SEQ ID NO: 35 (miRBase Accession No. MIMAT0000069) and other species homologs or orthologs.
- the hsa-miR-16-5p gene can be obtained by the method described in Lagos-Quintana M et al., 2001, Science, 294, p853-858.
- “hsa-miR-16-5p” has “hsa-mir-16-1”, “hsa-mir-16-2” (miRBase Accession No. MI00000070, MI0000115, which takes a hairpin-like structure as a precursor thereof.
- SEQ ID NOs: 184, 185) are known.
- hsa-miR-602 gene or “hsa-miR-602” refers to the hsa-miR-602 gene (miRBase Accession No. MIMAT0003270) described in SEQ ID NO: 36 or other species. Includes homologs or orthologs.
- the hsa-miR-602 gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci U S A, 103, p3687-3692.
- hsa-miR-602 “hsa-mir-602” (miRBase Accession No. MI0003615, SEQ ID NO: 186) having a hairpin-like structure as a precursor is known.
- hsa-miR-4651 gene or “hsa-miR-4651” refers to the hsa-miR-4651 gene (miRBase Accession No. MIMAT0019715) described in SEQ ID NO: 37 and other biological species. Includes homologs or orthologs.
- the hsa-miR-4651 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- “Hsa-miR-4651” is known as “hsa-mir-4651” (miRBase Accession No. MI0017279, SEQ ID NO: 187) having a hairpin-like structure as a precursor.
- hsa-miR-8069 gene or “hsa-miR-8069” refers to the hsa-miR-8069 gene (miRBase Accession No. MIMAT0030996) described in SEQ ID NO: 38 or other biological species. Includes homologs or orthologs.
- the hsa-miR-8069 gene can be obtained by the method described in Wang HJ et al., 2013, Shock, 39, p480-487.
- hsa-miR-8069 “hsa-mir-8069” (miRBase Accession No. MI0025905, SEQ ID NO: 188) having a hairpin-like structure as a precursor is known.
- hsa-miR-1238-5p gene or “hsa-miR-1238-5p” refers to the hsa-miR-1238-5p gene described in SEQ ID NO: 39 (miRBase Accession No. MIMAT0022947) and other species homologs or orthologs.
- the hsa-miR-1238-5p gene can be obtained by the method described in Berezikov E et al., 2007, Mol Cell, 28, p328-336.
- “hsa-miR-1238-5p” is known as “hsa-mir-1238” (miRBase Accession No. MI0006328, SEQ ID NO: 189) having a hairpin-like structure as a precursor.
- hsa-miR-6880-5p gene or “hsa-miR-6880-5p” refers to the hsa-miR-6880-5p gene described in SEQ ID NO: 40 (miRBase Accession No. MIMAT0027660) and other species homologs or orthologs.
- the hsa-miR-6880-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-6880-5p” is known as “hsa-mir-6880” (miRBase Accession No. MI0022727, SEQ ID NO: 190) having a hairpin-like structure as a precursor.
- hsa-miR-8072 gene or “hsa-miR-8072” refers to the hsa-miR-8072 gene (miRBase Accession No. MIMAT0030999) described in SEQ ID NO: 41 and other biological species. Includes homologs or orthologs.
- the hsa-miR-8072 gene can be obtained by the method described in Wang HJ et al., 2013, Shock, 39, p480-487.
- hsa-miR-8072 “hsa-mir-8072” (miRBase Accession No. MI0025908, SEQ ID NO: 191) having a hairpin-like structure as a precursor is known.
- hsa-miR-4723-5p gene or “hsa-miR-4723-5p” refers to the hsa-miR-4723-5p gene described in SEQ ID NO: 42 (miRBase Accession No. MIMAT0019838) and other species homologs or orthologs.
- the hsa-miR-4723-5p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- “hsa-miR-4723-5p” is known as “hsa-mir-4723” (miRBase Accession No. MI0017359, SEQ ID NO: 192) having a hairpin-like structure as a precursor.
- hsa-miR-4732-5p gene or “hsa-miR-4732-5p” refers to the hsa-miR-4732-5p gene (miRBase Accession No. 4) described in SEQ ID NO: 43. MIMAT0019855) and other species homologs or orthologs.
- the hsa-miR-4732-5p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- “Hsa-miR-4732-5p” is known as “hsa-mir-4732” (miRBase Accession No. MI0017369, SEQ ID NO: 193) having a hairpin-like structure as a precursor.
- hsa-miR-6125 gene or “hsa-miR-6125” refers to the hsa-miR-6125 gene (miRBase Accession No. MIMAT0024598) described in SEQ ID NO: 44 or other biological species. Includes homologs or orthologs.
- the hsa-miR-6125 gene can be obtained by the method described in Smith JL et al., 2012, J Virol, 86, p5278-5287.
- hsa-miR-6125 is known as “hsa-mir-6125” (miRBase Accession No. MI0021259, SEQ ID NO: 194) having a hairpin-like structure as a precursor.
- hsa-miR-6090 gene or “hsa-miR-6090” refers to the hsa-miR-6090 gene (miRBase Accession No. MIMAT0023715) described in SEQ ID NO: 45 and other biological species. Includes homologs or orthologs.
- the hsa-miR-6090 gene can be obtained by the method described in Yo JK et al., 2012, Stem Cells Dev, 21, p2049-2057.
- hsa-miR-6090 is known as “hsa-mir-6090” (miRBase Accession No. MI0020367, SEQ ID NO: 195) having a hairpin-like structure as a precursor.
- hsa-miR-7114-5p gene or “hsa-miR-7114-5p” refers to the hsa-miR-7114-5p gene described in SEQ ID NO: 46 (miRBase Accession No. MIMAT0028125) and other species homologs or orthologs.
- the hsa-miR-7114-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-7114-5p” is known as “hsa-mir-7114” (miRBase Accession No. MI0022965, SEQ ID NO: 196) having a hairpin-like structure as a precursor.
- hsa-miR-564 gene or “hsa-miR-564” refers to the hsa-miR-564 gene (miRBase Accession No. MIMAT0003228) described in SEQ ID NO: 47 and other biological species. Includes homologs or orthologs.
- the hsa-miR-564 gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci U S A, 103, p3687-3692.
- “hsa-miR-564” is known as “hsa-mir-564” (miRBase Accession No. MI0003570, SEQ ID NO: 197) having a hairpin-like structure as a precursor.
- hsa-miR-451a gene or “hsa-miR-451a” refers to the hsa-miR-451a gene (miRBase Accession No. MIMAT0001631) described in SEQ ID NO: 48 or other biological species. Includes homologs or orthologs.
- the hsa-miR-451a gene can be obtained by the method described in Altuvia Y et al., 2005, Nucleic Acids Res, 33, p2697-2706.
- hsa-miR-451a “hsa-mir-451a” (miRBase Accession No. MI0001729, SEQ ID NO: 198) having a hairpin-like structure as a precursor is known.
- hsa-miR-3135b gene or “hsa-miR-3135b” refers to the hsa-miR-3135b gene (miRBase Accession No. MIMAT0018985) described in SEQ ID NO: 49 or other species. Includes homologs or orthologs.
- the hsa-miR-3135b gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- “hsa-miR-3135b” is known as “hsa-mir-3135b” (miRBase Accession No. MI0016809, SEQ ID NO: 199) having a hairpin-like structure as a precursor.
- hsa-miR-4497 gene or “hsa-miR-4497” refers to the hsa-miR-4497 gene (miRBase Accession No. MIMAT0019032) described in SEQ ID NO: 50 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4497 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-mir-4497 (miRBase Accession No. MI0016859, SEQ ID NO: 200) having a hairpin-like structure as a precursor is known.
- hsa-miR-4665-5p gene or “hsa-miR-4665-5p” refers to the hsa-miR-4665-5p gene (miRBase Accession No. 5) described in SEQ ID NO: 51. MIMAT0019739) and other species homologs or orthologs.
- the hsa-miR-4665-5p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- “hsa-miR-4665-5p” “hsa-mir-4665” (miRBase Accession No. MI0017295, SEQ ID NO: 201) having a hairpin-like structure as a precursor is known.
- hsa-miR-3622a-5p gene or “hsa-miR-3622a-5p” refers to the hsa-miR-3622a-5p gene described in SEQ ID NO: 52 (miRBase Accession No. MIMAT0018003) and other species homologs or orthologs.
- the hsa-miR-3622a-5p gene can be obtained by the method described in Witten D et al., 2010, BMC Biol, 8, p58.
- “hsa-miR-3622a-5p” “hsa-mir-3622a” (miRBase Accession No. MI0016013, SEQ ID NO: 202) having a hairpin-like structure as a precursor is known.
- hsa-miR-6850-5p gene or “hsa-miR-6850-5p” refers to the hsa-miR-6850-5p gene described in SEQ ID NO: 53 (miRBase Accession No. MIMAT0027600) and other species homologs or orthologs.
- the hsa-miR-6850-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-mir-6850 (miRBase Accession No. MI0022696, SEQ ID NO: 203) having a hairpin-like structure as a precursor is known.
- hsa-miR-6821-5p gene or “hsa-miR-6821-5p” refers to the hsa-miR-6821-5p gene described in SEQ ID NO: 54 (miRBase Accession No. MIMAT0027542) and other species homologs or orthologs.
- the hsa-miR-6821-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-6821-5p” is known as “hsa-mir-6821” (miRBase Accession No. MI0022666, SEQ ID NO: 204) having a hairpin-like structure as a precursor.
- hsa-miR-5100 gene or “hsa-miR-5100” refers to the hsa-miR-5100 gene (miRBase Accession No. MIMAT0022259) described in SEQ ID NO: 55 or other biological species. Includes homologs or orthologs.
- the hsa-miR-5100 gene can be obtained by the method described in Tandon M et al., 2012, Oral Dis, 18, p127-131.
- “hsa-miR-5100” is known as “hsa-mir-5100” (miRBase Accession No. MI0019116, SEQ ID NO: 205) having a hairpin-like structure as a precursor.
- hsa-miR-6872-3p gene or “hsa-miR-6872-3p” refers to the hsa-miR-6872-3p gene described in SEQ ID NO: 56 (miRBase Accession No. MIMAT0027645) and other species homologs or orthologs.
- the hsa-miR-6872-3p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-6872-3p” is known as “hsa-mir-6872” (miRBase Accession No. MI0022719, SEQ ID NO: 206) having a hairpin-like structure as a precursor.
- hsa-miR-4433-3p gene or “hsa-miR-4433-3p” refers to the hsa-miR-4433-3p gene described in SEQ ID NO: 57 (miRBase Accession No. MIMAT0018949) and other species homologs or orthologs.
- the hsa-miR-4433-3p gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-mir-4433 miRBase Accession No. MI0016773, SEQ ID NO: 207 having a hairpin-like structure as a precursor is known.
- hsa-miR-1227-5p gene or “hsa-miR-1227-5p” refers to the hsa-miR-1227-5p gene described in SEQ ID NO: 58 (miRBase Accession No. MIMAT0022941) and other species homologs or orthologs.
- the hsa-miR-1227-5p gene can be obtained by the method described in Berezikov E et al., 2007, Mol Cell, 28, p328-336.
- hsa-miR-1227-5p “hsa-mir-1227” (miRBase Accession No. MI0006316, SEQ ID NO: 208) having a hairpin-like structure as a precursor is known.
- hsa-miR-3188 gene or “hsa-miR-3188” refers to the hsa-miR-3188 gene (miRBase Accession No. MIMAT0015070) described in SEQ ID NO: 59 or other biological species. Includes homologs or orthologs.
- the hsa-miR-3188 gene can be obtained by the method described in Stark MS et al., 2010, PLoS One, Vol. 5, e9685.
- hsa-miR-3188 “hsa-mir-3188” (miRBase Accession No. MI0014232, SEQ ID NO: 209) having a hairpin-like structure as a precursor is known.
- hsa-miR-7704 gene or “hsa-miR-7704” refers to the hsa-miR-7704 gene (miRBase Accession No. MIMAT0030019) described in SEQ ID NO: 60 or other biological species. Includes homologs or orthologs.
- the hsa-miR-7704 gene can be obtained by the method described in Swamithan S et al., 2013, Biochem Biophys Res Commun, 434, p228-234.
- hsa-miR-7704 “hsa-mir-7704” (miRBase Accession No. MI0025240, SEQ ID NO: 210) having a hairpin-like structure as a precursor is known.
- hsa-miR-3185 gene or “hsa-miR-3185” refers to the hsa-miR-3185 gene (miRBase Accession No. MIMAT0015065) described in SEQ ID NO: 61 or other species. Includes homologs or orthologs.
- the hsa-miR-3185 gene can be obtained by the method described in Stark MS et al., 2010, PLoS One, 5, e9685.
- hsa-miR-3185 “hsa-mir-3185” (miRBase Accession No. MI0014227, SEQ ID NO: 211) having a hairpin-like structure as a precursor is known.
- hsa-miR-1908-3p gene or “hsa-miR-1908-3p” refers to the hsa-miR-1908-3p gene described in SEQ ID NO: 62 (miRBase Accession No. MIMAT0026916) and other species homologs or orthologs.
- the hsa-miR-1908-3p gene can be obtained by the method described in Bar M et al., 2008, Stem Cells, 26, p2496-2505.
- “hsa-miR-1908-3p” is known as “hsa-mir-1908” (miRBase Accession No. MI0008329, SEQ ID NO: 212) having a hairpin-like structure as a precursor.
- hsa-miR-6781-5p gene or “hsa-miR-6781-5p” refers to the hsa-miR-6781-5p gene described in SEQ ID NO: 63 (miRBase Accession No. MIMAT0027462) and other species homologs or orthologs.
- the hsa-miR-6781-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-6781-5p” is known as “hsa-mir-6781” (miRBase Accession No. MI0022626, SEQ ID NO: 213) having a hairpin-like structure as a precursor.
- hsa-miR-6805-5p gene or “hsa-miR-6805-5p” refers to the hsa-miR-6805-5p gene described in SEQ ID NO: 64 (miRBase Accession No. MIMAT0027510) and other species homologs or orthologs.
- the hsa-miR-6805-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6805-5p “hsa-mir-6805” (miRBase Accession No. MI0022650, SEQ ID NO: 214) having a hairpin-like structure as a precursor is known.
- hsa-miR-8089 gene or “hsa-miR-8089” refers to the hsa-miR-8089 gene (miRBase Accession No. MIMAT0031016) described in SEQ ID NO: 65 or other biological species. Includes homologs or orthologs.
- the hsa-miR-8089 gene can be obtained by the method described in Wang HJ et al., 2013, Shock, 39, p480-487.
- “hsa-miR-8089” is known as “hsa-mir-8089” (miRBase Accession No. MI0025925, SEQ ID NO: 215) having a hairpin-like structure as a precursor.
- hsa-miR-665 gene or “hsa-miR-665” refers to the hsa-miR-665 gene (miRBase Accession No. MIMAT0004952) described in SEQ ID NO: 66 or other biological species. Includes homologs or orthologs.
- the hsa-miR-665 gene can be obtained by the method described in Berezikov E et al., 2006, Genome Res, 16, p1299-1298.
- hsa-miR-665 “hsa-mir-665” (miRBase Accession No. MI0005563, SEQ ID NO: 216) having a hairpin-like structure as a precursor is known.
- hsa-miR-4486 gene or “hsa-miR-4486” refers to the hsa-miR-4486 gene (miRBase Accession No. MIMAT0019020) described in SEQ ID NO: 67 and other biological species. Includes homologs or orthologs.
- the hsa-miR-4486 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-mir-4486 (miRBase Accession No. MI0016847, SEQ ID NO: 217) having a hairpin-like structure as a precursor is known.
- hsa-miR-6722-3p gene or “hsa-miR-6722-3p” refers to the hsa-miR-6722-3p gene described in SEQ ID NO: 68 (miRBase Accession No. MIMAT0025854) and other species homologs or orthologs.
- the hsa-miR-6722-3p gene can be obtained by the method described in Li Y et al., 2012, Gene, 497, p330-335.
- hsa-miR-6722-3p “hsa-mir-6722” (miRBase Accession No. MI0022557, SEQ ID NO: 218) having a hairpin-like structure as a precursor is known.
- hsa-miR-1260a gene or “hsa-miR-1260a” refers to the hsa-miR-1260a gene (miRBase Accession No. MIMAT0005911) described in SEQ ID NO: 69 or other species. Includes homologs or orthologs.
- the hsa-miR-1260a gene can be obtained by the method described in Morin RD et al., 2008, Genome Res, 18, p610-621.
- “hsa-miR-1260a” is known as “hsa-mir-1260a” (miRBase Accession No. MI0006394, SEQ ID NO: 219) having a hairpin-like structure as a precursor.
- hsa-miR-4707-5p gene or “hsa-miR-4707-5p” refers to the hsa-miR-4707-5p gene described in SEQ ID NO: 70 (miRBase Accession No. MIMAT0019807) and other species homologs or orthologs.
- the hsa-miR-4707-5p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- hsa-miR-4707-5p “hsa-mir-4707” (miRBase Accession No. MI0017340, SEQ ID NO: 220) having a hairpin-like structure as a precursor is known.
- hsa-miR-6741-5p gene or “hsa-miR-6741-5p” refers to the hsa-miR-6741-5p gene described in SEQ ID NO: 71 (miRBase Accession No. MIMAT0027383) and other species homologs or orthologs.
- the hsa-miR-6741-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6741-5p “hsa-mir-6741” (miRBase Accession No. MI0022586, SEQ ID NO: 221) having a hairpin-like structure as a precursor is known.
- hsa-miR-1260b gene or “hsa-miR-1260b” refers to the hsa-miR-1260b gene (miRBase Accession No. MIMAT0015041) described in SEQ ID NO: 72 or other biological species. Includes homologs or orthologs.
- the hsa-miR-1260b gene can be obtained by the method described in Stark MS et al., 2010, PLoS One, Vol. 5, e9685.
- hsa-miR-1260b “hsa-mir-1260b” (miRBase Accession No. MI0014197, SEQ ID NO: 222) having a hairpin-like structure as a precursor is known.
- hsa-miR-1246 gene or “hsa-miR-1246” refers to the hsa-miR-1246 gene (miRBase Accession No. MIMAT0005898) described in SEQ ID NO: 73 and other biological species. Includes homologs or orthologs.
- the hsa-miR-1246 gene can be obtained by the method described in Morin RD et al., 2008, Genome Res, 18, p610-621.
- hsa-miR-1246 “hsa-mir-1246” (miRBase Accession No. MI0006381, SEQ ID NO: 223) having a hairpin-like structure as a precursor is known.
- hsa-miR-6845-5p gene or “hsa-miR-6845-5p” refers to the hsa-miR-6845-5p gene described in SEQ ID NO: 74 (miRBase Accession No. MIMAT0027590) and other species homologs or orthologs.
- the hsa-miR-6845-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6845-5p “hsa-mir-6845” (miRBase Accession No. MI0022691, SEQ ID NO: 224) having a hairpin-like structure as a precursor is known.
- hsa-miR-4638-5p gene or “hsa-miR-4638-5p” refers to the hsa-miR-4638-5p gene described in SEQ ID NO: 75 (miRBase Accession No. MIMAT0019695) and other species homologs or orthologs.
- the hsa-miR-4638-5p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- “hsa-miR-4638-5p” “hsa-mir-4638” (miRBase Accession No. MI0017265, SEQ ID NO: 225) having a hairpin-like structure as a precursor is known.
- hsa-miR-6085 gene or “hsa-miR-6085” refers to the hsa-miR-6085 gene (miRBase Accession No. MIMAT0023710) described in SEQ ID NO: 76 or other biological species. Includes homologs or orthologs.
- the hsa-miR-6085 gene can be obtained by the method described in Voellenkle C et al., 2012, RNA, 18, p472-484.
- hsa-miR-6085 “hsa-mir-6085” (miRBase Accession No. MI0020362, SEQ ID NO: 226) having a hairpin-like structure as a precursor is known.
- hsa-miR-1228-3p gene or “hsa-miR-1228-3p” refers to the hsa-miR-1228-3p gene described in SEQ ID NO: 77 (miRBase Accession No. MIMAT0005583) and other species homologs or orthologs.
- the hsa-miR-1228-3p gene can be obtained by the method described in Berezikov E et al., 2007, Mol Cell, 28, p328-336.
- hsa-mir-1228 (miRBase Accession No. MI0006318, SEQ ID NO: 227) having a hairpin-like structure as a precursor is known.
- hsa-miR-4534 gene or “hsa-miR-4534” refers to the hsa-miR-4534 gene (miRBase Accession No. MIMAT0019073) described in SEQ ID NO: 78 and other biological species. Includes homologs or orthologs.
- the hsa-miR-4534 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- “hsa-miR-4534” is known as “hsa-mir-4534” (miRBase Accession No. MI0016901, SEQ ID NO: 228) having a hairpin-like structure as a precursor.
- hsa-miR-5585-3p gene or “hsa-miR-5585-3p” refers to the hsa-miR-5585-3p gene described in SEQ ID NO: 79 (miRBase Accession No. MIMAT0022286) and other species homologs or orthologs.
- the hsa-miR-5585-3p gene can be obtained by the method described in Friedlander® MR et al., 2012, Nucleic® Acids® Res, 40, p37-52.
- hsa-mir-5585 miRBase Accession No. MI0019142, SEQ ID NO: 229 having a hairpin-like structure as a precursor is known.
- hsa-miR-4741 gene or “hsa-miR-4741” refers to the hsa-miR-4741 gene (miRBase Accession No. MIMAT0019871) described in SEQ ID NO: 80 Includes homologs or orthologs.
- the hsa-miR-4741 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, 71, p78-86.
- hsa-miR-4741 “hsa-mir-4741” (miRBase Accession No. MI0017379, SEQ ID NO: 230) having a hairpin-like structure as a precursor is known.
- hsa-miR-4433b-3p gene or “hsa-miR-4433b-3p” refer to the hsa-miR-4433b-3p gene described in SEQ ID NO: 81 (miRBase Accession No. MIMAT0030414) and other species homologs or orthologs.
- the hsa-miR-4433b-3p gene can be obtained by the method described in Ple H et al., 2012, PLoS One, Vol. 7, e50746.
- hsa-mir-4433b miRBase Accession No. MI0025511, SEQ ID NO: 231 having a hairpin-like structure as a precursor is known.
- hsa-miR-197-5p gene or “hsa-miR-197-5p” refers to the hsa-miR-197-5p gene described in SEQ ID NO: 82 (miRBase Accession No. MIMAT0022691) and other species homologs or orthologs.
- the hsa-miR-197-5p gene can be obtained by the method described in Lagos-Quintana M et al., 2003, RNA, Vol. 9, p175-179.
- “hsa-miR-197-5p” “hsa-mir-197” (miRBase Accession No. MI0000239, SEQ ID NO: 232) having a hairpin-like structure as a precursor is known.
- hsa-miR-718 gene or “hsa-miR-718” refers to the hsa-miR-718 gene (miRBase Accession No. MIMAT0012735) described in SEQ ID NO: 83 or other biological species. Includes homologs or orthologs.
- the hsa-miR-718 gene can be obtained by the method described in Artzi S et al., 2008, BMC Bioinformatics, Vol. 9, p39.
- “hsa-miR-718” is known as “hsa-mir-718” (miRBase Accession No. MI0012489, SEQ ID NO: 233) having a hairpin-like structure as a precursor.
- hsa-miR-4513 gene or “hsa-miR-4513” refers to the hsa-miR-4513 gene (miRBase Accession No. MIMAT0019050) described in SEQ ID NO: 84 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4513 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-mir-4513 (miRBase Accession No. MI0016879, SEQ ID NO: 234) having a hairpin-like structure as a precursor is known.
- hsa-miR-4446-3p gene or “hsa-miR-4446-3p” refers to the hsa-miR-4446-3p gene described in SEQ ID NO: 85 (miRBase Accession No. MIMAT0018965) and other species homologs or orthologs.
- the hsa-miR-4446-3p gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- “hsa-miR-4446-3p” is known as “hsa-mir-4446” (miRBase Accession No. MI0016789, SEQ ID NO: 235) having a hairpin-like structure as a precursor.
- hsa-miR-619-5p gene or “hsa-miR-619-5p” refers to the hsa-miR-619-5p gene described in SEQ ID NO: 86 (miRBase Accession No. MIMAT0026622) and other species homologs or orthologs.
- the hsa-miR-619-5p gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci U S A, 103, p3687-3692.
- “hsa-miR-619-5p” is known as “hsa-mir-619” (miRBase Accession No. MI0003633, SEQ ID NO: 236) having a hairpin-like structure as a precursor.
- hsa-miR-6816-5p gene or “hsa-miR-6816-5p” refers to the hsa-miR-6816-5p gene described in SEQ ID NO: 87 (miRBase Accession No. MIMAT0027532) and other species homologs or orthologs.
- the hsa-miR-6816-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6816-5p “hsa-mir-6816” (miRBase Accession No. MI0022661, SEQ ID NO: 237) having a hairpin-like structure as a precursor is known.
- hsa-miR-6778-5p gene or “hsa-miR-6778-5p” refers to the hsa-miR-6778-5p gene described in SEQ ID NO: 88 (miRBase Accession No. MIMAT0027456) and other species homologs or orthologs.
- the hsa-miR-6778-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6778-5p “hsa-mir-6778” (miRBase Accession No. MI0022623, SEQ ID NO: 238) having a hairpin-like structure as a precursor is known.
- hsa-miR-24-3p gene or “hsa-miR-24-3p” refer to the hsa-miR-24-3p gene described in SEQ ID NO: 89 (miRBase Accession No. MIMAT00000080) and other species homologues or orthologues.
- the hsa-miR-24-3p gene can be obtained by the method described in Lagos-Quintana M et al., 2001, Science, 294, p853-858.
- “Hsa-miR-24-3p” has a hairpin-like structure as its precursor, “hsa-mir-24-1,” “hsa-mir-24-2” (miRBase Accession No. MI00000080, MI00000081, SEQ ID NOs: 239, 240) are known.
- hsa-miR-1915-3p gene or “hsa-miR-1915-3p” refers to the hsa-miR-1915-3p gene described in SEQ ID NO: 90 (miRBase Accession No. MIMAT0007892) and other species homologs or orthologs.
- the hsa-miR-1915-3p gene can be obtained by the method described in Bar M et al., 2008, Stem Cells, 26, p2496-2505.
- hsa-miR-1915-3p “hsa-mir-1915” (miRBase Accession No. MI0008336, SEQ ID NO: 241) having a hairpin-like structure as a precursor is known.
- hsa-miR-4665-3p gene or “hsa-miR-4665-3p” refers to the hsa-miR-4665-3p gene described in SEQ ID NO: 91 (miRBase Accession No. MIMAT0019740) and other species homologs or orthologs.
- the hsa-miR-4665-3p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- hsa-miR-4665-3p “hsa-mir-4665” (miRBase Accession No. MI0017295, SEQ ID NO: 201) having a hairpin-like structure as a precursor is known.
- hsa-miR-4449 gene or “hsa-miR-4449” refers to the hsa-miR-4449 gene (miRBase Accession No. MIMAT0018968) described in SEQ ID NO: 92 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4449 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-mir-4449 (miRBase Accession No. MI0016792, SEQ ID NO: 242) having a hairpin-like structure as a precursor is known.
- hsa-miR-6889-5p gene or “hsa-miR-6889-5p” refers to the hsa-miR-6889-5p gene described in SEQ ID NO: 93 (miRBase Accession No. MIMAT0027678) and other species homologs or orthologs.
- the hsa-miR-6889-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-6889-5p” is known as “hsa-mir-6889” (miRBase Accession No. MI0022736, SEQ ID NO: 243) having a hairpin-like structure as a precursor.
- hsa-miR-486-3p gene or “hsa-miR-486-3p” refer to the hsa-miR-486-3p gene described in SEQ ID NO: 94 (miRBase Accession No. MIMAT0004762) and other species homologs or orthologs.
- the hsa-miR-486-3p gene can be obtained by the method described in Fu H et al., 2005, FEBS Lett, 579, p3849-3854.
- “hsa-miR-486-3p” has “hsa-mir-486, hsa-mir-486-2” (miRBase Accession No. 245) is known.
- hsa-miR-7113-3p gene or “hsa-miR-7113-3p” refers to the hsa-miR-7113-3p gene described in SEQ ID NO: 95 (miRBase Accession No. MIMAT0028124) and other species homologs or orthologs.
- the hsa-miR-7113-3p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-7113-3p “hsa-mir-7113” (miRBase Accession No. MI0022964, SEQ ID NO: 246) having a hairpin-like structure as a precursor is known.
- hsa-miR-642a-3p gene or “hsa-miR-642a-3p” refers to the hsa-miR-642a-3p gene described in SEQ ID NO: 96 (miRBase Accession No. MIMAT0020924) and other species homologs or orthologs.
- the hsa-miR-642a-3p gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci U S A, 103, p3687-3692.
- “hsa-miR-642a-3p” is known as “hsa-mir-642a” (miRBase Accession No. MI0003657, SEQ ID NO: 247) having a hairpin-like structure as a precursor.
- hsa-miR-7847-3p gene or “hsa-miR-7847-3p” refers to the hsa-miR-7847-3p gene described in SEQ ID NO: 97 (miRBase Accession No. MIMAT0030422) and other species homologs or orthologs.
- the hsa-miR-7847-3p gene can be obtained by the method described in Ple H et al., 2012, PLoS One, Vol. 7, e50746.
- “hsa-miR-7847-3p” is known as “hsa-mir-7847” (miRBase Accession No. MI0025517, SEQ ID NO: 248) having a hairpin-like structure as a precursor.
- hsa-miR-6768-5p gene or “hsa-miR-6768-5p” refers to the hsa-miR-6768-5p gene described in SEQ ID NO: 98 (miRBase Accession No. MIMAT0027436) and other species homologs or orthologs.
- the hsa-miR-6768-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6768-5p “hsa-mir-6768” (miRBase Accession No. MI0022613, SEQ ID NO: 249) having a hairpin-like structure as a precursor is known.
- hsa-miR-1290 gene or “hsa-miR-1290” refers to the hsa-miR-1290 gene (miRBase Accession No. MIMAT0005880) described in SEQ ID NO: 99 and other biological species. Includes homologs or orthologs.
- the hsa-miR-1290 gene can be obtained by the method described in Morin RD et al., 2008, Genome Res, 18, p610-621.
- hsa-miR-1290 “hsa-mir-1290” (miRBase Accession No. MI0006352, SEQ ID NO: 250) having a hairpin-like structure as a precursor is known.
- hsa-miR-7108-5p gene or “hsa-miR-7108-5p” refers to the hsa-miR-7108-5p gene described in SEQ ID NO: 100 (miRBase Accession No. MIMAT0028113) and other species homologs or orthologs.
- the hsa-miR-7108-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-7108-5p” is known as “hsa-mir-7108” (miRBase Accession No. MI0022959, SEQ ID NO: 251) having a hairpin-like structure as a precursor.
- hsa-miR-92b-5p gene or “hsa-miR-92b-5p” refers to the hsa-miR-92b-5p gene described in SEQ ID NO: 101 (miRBase Accession No. MIMAT0004792) and other species homologs or orthologs.
- the hsa-miR-92b-5p gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci U S A, 103, p3687-3692.
- hsa-miR-92b-5p “hsa-mir-92b” (miRBase Accession No. MI0003560, SEQ ID NO: 252) having a hairpin-like structure as a precursor is known.
- hsa-miR-663b gene or “hsa-miR-663b” refers to the hsa-miR-663b gene (miRBase Accession No. MIMAT0005867) described in SEQ ID NO: 102 or other biological species. Includes homologs or orthologs.
- the hsa-miR-663b gene can be obtained by the method described in Takada S et al., 2008, Leukemia, Vol. 22, p1274-1278.
- hsa-miR-663b is known as “hsa-mir-663b” (miRBase Accession No. MI0006336, SEQ ID NO: 253) having a hairpin-like structure as a precursor.
- hsa-miR-3940-5p gene or “hsa-miR-3940-5p” refers to the hsa-miR-3940-5p gene described in SEQ ID NO: 103 (miRBase Accession No. MIMAT0019229) and other species homologs or orthologs.
- the hsa-miR-3940-5p gene can be obtained by the method described in Liao JY et al., 2010, PLoS One, 5, e10563.
- “hsa-miR-3940-5p” is known as “hsa-mir-3940” (miRBase Accession No. MI0016597, SEQ ID NO: 254) having a hairpin-like structure as a precursor.
- hsa-miR-4467 gene or “hsa-miR-4467” refers to the hsa-miR-4467 gene (miRBase Accession No. MIMAT0018994) described in SEQ ID NO: 104 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4467 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-mir-4467 (miRBase Accession No. MI0016818, SEQ ID NO: 255) having a hairpin-like structure as a precursor is known.
- hsa-miR-6858-5p gene or “hsa-miR-6858-5p” refers to the hsa-miR-6858-5p gene described in SEQ ID NO: 105 (miRBase Accession No. MIMAT0027616) and other species homologs or orthologs.
- the hsa-miR-6858-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-6858-5p “hsa-mir-6858” (miRBase Accession No. MI0022704, SEQ ID NO: 256) having a hairpin-like structure as a precursor is known.
- hsa-miR-4417 gene or “hsa-miR-4417” refers to the hsa-miR-4417 gene (miRBase Accession No. MIMAT0018929) described in SEQ ID NO: 106 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4417 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-miR-4417 “hsa-mir-4417” (miRBase Accession No. MI0016753, SEQ ID NO: 257) having a hairpin-like structure as a precursor is known.
- hsa-miR-3665 gene or “hsa-miR-3665” refers to the hsa-miR-3665 gene (miRBase Accession No. MIMAT0018087) described in SEQ ID NO: 107 and other biological species. Includes homologs or orthologs.
- the hsa-miR-3665 gene can be obtained by the method described in Xie X et al., 2005, Nature, 434, p338-345.
- hsa-miR-3665 “hsa-mir-3665” (miRBase Accession No. MI0016066, SEQ ID NO: 258) having a hairpin-like structure as a precursor is known.
- hsa-miR-4736 gene or “hsa-miR-4736” refers to the hsa-miR-4736 gene (miRBase Accession No. MIMAT0019862) described in SEQ ID NO: 108 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4736 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, 71, p78-86.
- hsa-miR-4736 “hsa-mir-4736” (miRBase Accession No. MI0017373, SEQ ID NO: 259) having a hairpin-like structure as a precursor is known.
- hsa-miR-4687-3p gene or “hsa-miR-4687-3p” refers to the hsa-miR-4687-3p gene described in SEQ ID NO: 109 (miRBase Accession No. MIMAT0019775) and other species homologs or orthologs.
- the hsa-miR-4687-3p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- “hsa-miR-4687-3p” “hsa-mir-4687” (miRBase Accession No. MI0017319, SEQ ID NO: 260) having a hairpin-like structure as a precursor is known.
- hsa-miR-1908-5p gene or “hsa-miR-1908-5p” refers to the hsa-miR-1908-5p gene described in SEQ ID NO: 110 (miRBase Accession No. MIMAT0007881) and other species homologs or orthologs.
- the hsa-miR-1908-5p gene can be obtained by the method described in Bar M et al., 2008, Stem Cells, 26, p2496-5055.
- “Hsa-miR-1908-5p” is known as “hsa-mir-1908” (miRBase Accession No. MI0008329, SEQ ID NO: 212), which has a hairpin-like structure as a precursor.
- hsa-miR-5195-3p gene or “hsa-miR-5195-3p” refers to the hsa-miR-5195-3p gene described in SEQ ID NO: 111 (miRBase Accession No. MIMAT0021127) and other species homologs or orthologs.
- the hsa-miR-5195-3p gene can be obtained by the method described in Schott D et al., 2011, Leukemia, 25, p1389-1399.
- hsa-miR-5195-3p “hsa-mir-5195” (miRBase Accession No. MI0018174, SEQ ID NO: 261) having a hairpin-like structure as a precursor is known.
- hsa-miR-4286 gene or “hsa-miR-4286” refers to the hsa-miR-4286 gene (miRBase Accession No. MIMAT0016916) described in SEQ ID NO: 112 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4286 gene can be obtained by the method described in Goff LA et al., 2009, PLoS One, vol. 4, e7192.
- hsa-miR-4286 is known as “hsa-mir-4286” (miRBase Accession No. MI0015894, SEQ ID NO: 262) having a hairpin-like structure as a precursor.
- hsa-miR-3679-3p gene or “hsa-miR-3679-3p” refers to the hsa-miR-3679-3p gene described in SEQ ID NO: 113 (miRBase Accession No. MIMAT0018105) and other species homologs or orthologs.
- the hsa-miR-3679-3p gene can be obtained by the method described in Creighton CJ et al., 2010, PLoS One, 5, e9637.
- hsa-miR-3679-3p “hsa-mir-3679” (miRBase Accession No. MI0016080, SEQ ID NO: 263) having a hairpin-like structure as a precursor is known.
- hsa-miR-6791-5p gene or “hsa-miR-6791-5p” refers to the hsa-miR-6791-5p gene described in SEQ ID NO: 114 (miRBase Accession No. MIMAT0027482) and other species homologs or orthologs.
- the hsa-miR-6791-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-6791-5p” is known as “hsa-mir-6791” (miRBase Accession No. MI0022636, SEQ ID NO: 264) having a hairpin-like structure as a precursor.
- hsa-miR-1202 gene or “hsa-miR-1202” refers to the hsa-miR- 1202 gene (miRBase Accession No. MIMAT0005865) described in SEQ ID NO: 115 and other species. Includes homologs or orthologs.
- the hsa-miR- 1202 gene can be obtained by the method described in Martin S et al., 2008, Leukemia, 22, p330-338.
- hsa-miR-1220 is known as “hsa-mir-1220” (miRBase Accession No. MI0006334, SEQ ID NO: 265) having a hairpin-like structure as a precursor.
- hsa-miR-3656 gene or “hsa-miR-3656” refers to the hsa-miR-3656 gene (miRBase Accession No. MIMAT0018076) described in SEQ ID NO: 116 or other species. Includes homologs or orthologs.
- the hsa-miR-3656 gene can be obtained by the method described in Meiri E et al., 2010, Nucleic Acids Res, 38, p6234-6246.
- hsa-miR-3656 “hsa-mir-3656” (miRBase Accession No. MI0016056, SEQ ID NO: 266) having a hairpin-like structure as a precursor is known.
- hsa-miR-4746-3p gene or “hsa-miR-4746-3p” refers to the hsa-miR-4746-3p gene described in SEQ ID NO: 117 (miRBase Accession No. MIMAT0019881) and other species homologs or orthologs.
- the hsa-miR-4746-3p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- hsa-miR-4746-3p “hsa-mir-4746” (miRBase Accession No. MI0017385, SEQ ID NO: 267) having a hairpin-like structure as a precursor is known.
- hsa-miR-3184-5p gene or “hsa-miR-3184-5p” refers to the hsa-miR-3184-5p gene described in SEQ ID NO: 118 (miRBase Accession No. MIMAT0015064) and other species homologs or orthologs.
- the hsa-miR-3184-5p gene can be obtained by the method described in Stark MS et al. 2010, PLoS One, Vol. 5, e9685.
- “hsa-miR-3184-5p” is known as “hsa-mir-3184” (miRBase Accession No. MI0014226, SEQ ID NO: 268) having a hairpin-like structure as a precursor.
- hsa-miR-3937 gene or “hsa-miR-3937” refers to the hsa-miR-3937 gene (miRBase Accession No. MIMAT0018352) described in SEQ ID NO: 119 or other biological species. Includes homologs or orthologs.
- the hsa-miR-3937 gene can be obtained by the method described in Liao JY et al., 2010, PLoS One, 5, e10563.
- hsa-miR-3937 “hsa-mir-3937” (miRBase Accession No. MI0016593, SEQ ID NO: 269) having a hairpin-like structure as a precursor is known.
- hsa-miR-6515-3p gene or “hsa-miR-6515-3p” refers to the hsa-miR-6515-3p gene described in SEQ ID NO: 120 (miRBase Accession No. MIMAT0025487) and other species homologs or orthologs.
- the hsa-miR-6515-3p gene can be obtained by the method described in Joyce CE et al., 2011, Hum Mole Genet, 20, p4025-4040.
- “hsa-miR-6515-3p” is known as “hsa-mir-6515” (miRBase Accession No. MI0022227, SEQ ID NO: 270) having a hairpin-like structure as a precursor.
- hsa-miR-6132 gene or “hsa-miR-6132” refers to the hsa-miR-6132 gene (miRBase Accession No. MIMAT0024616) described in SEQ ID NO: 121 or other biological species. Includes homologs or orthologs.
- the hsa-miR-6132 gene can be obtained by the method described in Dannemann M et al., 2012, Genome Biol Evol, 4, p552-564.
- hsa-miR-6132 “hsa-mir-6132” (miRBase Accession No. MI0021277, SEQ ID NO: 271) having a hairpin-like structure as a precursor is known.
- hsa-miR-187-5p gene or “hsa-miR-187-5p” refers to the hsa-miR-187-5p gene described in SEQ ID NO: 122 (miRBase Accession No. MIMAT0004561) and other species homologs or orthologs.
- the hsa-miR-187-5p gene can be obtained by the method described in Lim LP et al., 2003, Science, 299, p1540.
- hsa-miR-187-5p “hsa-mir-187” (miRBase Accession No. MI000000274, SEQ ID NO: 272) having a hairpin-like structure as a precursor is known.
- hsa-miR-7111-5p gene or “hsa-miR-7111-5p” refers to the hsa-miR-7111-5p gene described in SEQ ID NO: 123 (miRBase Accession No. MIMAT0028119) and other species homologs or orthologs.
- the hsa-miR-7111-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- hsa-miR-7111-5p “hsa-mir-7111” (miRBase Accession No. MI0022962, SEQ ID NO: 273) having a hairpin-like structure as a precursor is known.
- hsa-miR-5787 gene or “hsa-miR-5787” refers to the hsa-miR-5787 gene (miRBase Accession No. MIMAT0023252) described in SEQ ID NO: 124 or other biological species. Includes homologs or orthologs.
- the hsa-miR-5787 gene can be obtained by the method described in Yo H et al., 2011, Biochem Biophys Res Commun, 415, p567-572.
- hsa-miR-5787 “hsa-mir-5787” (miRBase Accession No. MI0019797, SEQ ID NO: 274) having a hairpin-like structure as a precursor is known.
- hsa-miR-6679-5p gene or “hsa-miR-6679-5p” refers to the hsa-miR-6679-5p gene (miRBase Accession No. MIMAT0027458) and other species homologs or orthologs.
- the hsa-miR-6679-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “Hsa-miR-6779-5p” is known as “hsa-mir-6679” (miRBase Accession No. MI0022624, SEQ ID NO: 275) having a hairpin-like structure as a precursor.
- hsa-miR-6808-5p gene or “hsa-miR-6808-5p” refers to the hsa-miR-6808-5p gene described in SEQ ID NO: 126 (miRBase Accession No. MIMAT0027516) and other species homologs or orthologs.
- the hsa-miR-6808-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p1634-1645.
- hsa-miR-6808-5p “hsa-mir-6808” (miRBase Accession No. MI0022653, SEQ ID NO: 276) having a hairpin-like structure as a precursor is known.
- hsa-miR-6774-5p gene or “hsa-miR-6774-5p” refers to the hsa-miR-6774-5p gene described in SEQ ID NO: 127 (miRBase Accession No. MIMAT0027448) and other species homologs or orthologs.
- the hsa-miR-6774-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, 22, p1634-1645.
- “hsa-miR-6774-5p” “hsa-mir-6774” (miRBase Accession No. MI0022619, SEQ ID NO: 277) having a hairpin-like structure as a precursor is known.
- hsa-miR-4656 gene or “hsa-miR-4656” refers to the hsa-miR-4656 gene (miRBase Accession No. MIMAT0019723) described in SEQ ID NO: 128 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4656 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- hsa-miR-4656 “hsa-mir-4656” (miRBase Accession No. MI0017284, SEQ ID NO: 278) having a hairpin-like structure as a precursor is known.
- hsa-miR-6806-5p gene or “hsa-miR-6806-5p” refers to the hsa-miR-6806-5p gene described in SEQ ID NO: 129 (miRBase Accession No. 1). MIMAT0027512) and other species homologs or orthologs.
- the hsa-miR-6806-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p1634-1645.
- “hsa-miR-6806-5p” is known as “hsa-mir-6806” (miRBase Accession No. MI0022651, SEQ ID NO: 279) having a hairpin-like structure as a precursor.
- hsa-miR-1233-5p gene or “hsa-miR-1233-5p” refers to the hsa-miR-1233-5p gene (miRBase Accession No. MIMAT0022943) and other species homologs or orthologs.
- the hsa-miR-1233-5p gene can be obtained by the method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, p328-336.
- “hsa-miR-1233-5p” has a hairpin-like structure as a precursor thereof, “hsa-mir-1233-1,” “hsa-mir-12233-2” (miRBase Accession No. MI0006323, MI0015973, Sequence numbers 280, 281) are known.
- hsa-miR-328-5p gene or “hsa-miR-328-5p” refers to the hsa-miR-328-5p gene described in SEQ ID NO: 131 (miRBase Accession No. MIMAT0026486) and other species homologs or orthologs.
- the hsa-miR-328-5p gene can be obtained by the method described in Kim J et al., 2004, Proc Natl Acad Sci U S A, 101, p360-365.
- “hsa-miR-328-5p” “hsa-mir-328” (miRBase Accession No. MI000004, SEQ ID NO: 282) having a hairpin-like structure as a precursor is known.
- hsa-miR-4675 gene or “hsa-miR-4675” refers to the hsa-miR-4675 gene (miRBase Accession No. MIMAT0019756) described in SEQ ID NO: 132 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4675 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., 71, p78-86.
- “hsa-miR-4673” is known as “hsa-mir-4675” (miRBase Accession No. MI0017305, SEQ ID NO: 283) having a hairpin-like structure as a precursor.
- hsa-miR-2110 gene refers to the hsa-miR-2110 gene (miRBase Accession No. MIMAT0010133) described in SEQ ID NO: 133 and other species. Includes homologs or orthologs.
- the hsa-miR-2110 gene can be obtained by the method described in Zhu JY et al., 2009, J Virol, 83, p3333-3341.
- hsa-miR-2110 “hsa-mir-2110” (miRBase Accession No. MI0010629, SEQ ID NO: 284) having a hairpin-like structure as a precursor is known.
- hsa-miR-6076 gene or “hsa-miR-6076” refers to the hsa-miR-6076 gene (miRBase Accession No. MIMAT0023701) described in SEQ ID NO: 134 or other biological species. Includes homologs or orthologs.
- the hsa-miR-6076 gene can be obtained by the method described in Voellenkle C et al., 2012, RNA, 18, p472-484.
- hsa-miR-6076 “hsa-mir-6076” (miRBase Accession No. MI0020353, SEQ ID NO: 285) having a hairpin-like structure as a precursor is known.
- hsa-miR-3619-3p gene or “hsa-miR-3619-3p” refers to the hsa-miR-3619-3p gene described in SEQ ID NO: 135 (miRBase Accession No. MIMAT0019219) and other species homologs or orthologs.
- the hsa-miR-3619-3p gene can be obtained by the method described in Witten D et al., 2010, BMC Biol, 8, p58.
- “hsa-miR-3619-3p” “hsa-mir-3619” (miRBase Accession No. MI0016009, SEQ ID NO: 286) having a hairpin-like structure as a precursor is known.
- hsa-miR-92a-2-5p gene or “hsa-miR-92a-2-5p” refers to the hsa-miR-92a-2-5p described in SEQ ID NO: 136. It includes genes (miRBase Accession No. MIMAT0004508) and other species homologues or orthologues.
- the hsa-miR-92a-2-5p gene can be obtained by the method described in Mouretos Z et al., 2002, Genes Dev, 16, p720-728.
- “hsa-miR-92a-2-5p” is known as “hsa-mir-92a-2” (miRBase Accession No. MI00000094, SEQ ID NO: 287) having a hairpin-like structure as a precursor.
- hsa-miR-128-1-5p gene or “hsa-miR-128-1-5p” refers to the hsa-miR-128-1-5p set forth in SEQ ID NO: 137. It includes genes (miRBase Accession No. MIMAT0026477) and other species homologues or orthologues.
- the hsa-miR-128-1-5p gene can be obtained by the method described in Lagos-Quintana M et al., 2002, Curr Biol, 12, p735-739.
- “hsa-miR-128-1-5p” is known as “hsa-mir-128-1” (miRBase Accession No. MI0000447, SEQ ID NO: 288) having a hairpin-like structure as a precursor.
- hsa-miR-638 gene or “hsa-miR-638” refers to the hsa-miR-638 gene (miRBase Accession No. MIMAT0003308) described in SEQ ID NO: 138 or other biological species. Includes homologs or orthologs.
- the hsa-miR-638 gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci U S A, 103, p3687-3692.
- hsa-miR-638 “hsa-mir-638” (miRBase Accession No. MI0003653, SEQ ID NO: 289) having a hairpin-like structure as a precursor is known.
- hsa-miR-2861 gene or “hsa-miR-2861” refers to the hsa-miR-2861 gene (miRBase Accession No. MIMAT0013802) described in SEQ ID NO: 139 or other biological species. Includes homologs or orthologs.
- the hsa-miR-2861 gene can be obtained by the method described in Li H et al., 2009, J Clin Invest, 119, p3666-3777.
- hsa-miR-2861 “hsa-mir-2861” (miRBase Accession No. MI0013006, SEQ ID NO: 290) having a hairpin-like structure as a precursor is known.
- hsa-miR-371a-5p gene or “hsa-miR-371a-5p” refers to the hsa-miR-371a-5p gene described in SEQ ID NO: 140 (miRBase Accession No. MIMAT0004687) and other species homologs or orthologs.
- the hsa-miR-371a-5p gene can be obtained by the method described in Suh MR et al., 2004, Dev Biol, 270, p488-498.
- hsa-miR-371a-5p “hsa-mir-371a” (miRBase Accession No. MI000079, SEQ ID NO: 291) having a hairpin-like structure as a precursor is known.
- hsa-miR-211-3p gene or “hsa-miR-211-3p” refers to the hsa-miR-211-3p gene described in SEQ ID NO: 141 (miRBase Accession No. MIMAT0022694) and other species homologs or orthologs.
- the hsa-miR-211-3p gene can be obtained by the method described in Lim LP et al., 2003, Science, 299, p1540.
- “hsa-miR-211-3p” is known as “hsa-mir-211” (miRBase Accession No. MI0000287, SEQ ID NO: 292) having a hairpin-like structure as a precursor.
- hsa-miR-1273g-3p gene or “hsa-miR-1273g-3p” refers to the hsa-miR-1273g-3p gene described in SEQ ID NO: 142 (miRBase Accession No. MIMAT0022742) and other species homologs or orthologs.
- the hsa-miR-1273g-3p gene can be obtained by the method described in Reshmi G et al., 2011, Genomics, 97, p333-340.
- hsa-miR-1273g-3p “hsa-mir-1273g” (miRBase Accession No. MI0018003, SEQ ID NO: 293) having a hairpin-like structure as a precursor is known.
- hsa-miR-1203 gene or “hsa-miR-1203” refers to the hsa-miR-1203 gene (miRBase Accession No. MIMAT0005866) described in SEQ ID NO: 143 or other species. Includes homologs or orthologs.
- the hsa-miR-1203 gene can be obtained by the method described in Martin S et al., 2008, Leukemia, 22, p330-338.
- “Hsa-miR-1203” is known as “hsa-mir-1203” (miRBase Accession No. MI0006335, SEQ ID NO: 294), which has a hairpin-like structure as a precursor.
- hsa-miR-122-5p gene or “hsa-miR-122-5p” refers to the hsa-miR-122-5p gene described in SEQ ID NO: 144 (miRBase Accession No. MIMAT000021) and other species homologs or orthologs.
- the hsa-miR-122-5p gene can be obtained by the method described in Lagos-Quintana M et al., 2002, Curr Biol, 12, p735-739.
- “hsa-miR-122-5p” is known as “hsa-mir-122” (miRBase Accession No. MI000042, SEQ ID NO: 295) having a hairpin-like structure as a precursor.
- hsa-miR-4258 gene or “hsa-miR-4258” refers to the hsa-miR-4258 gene (miRBase Accession No. MIMAT0016879) described in SEQ ID NO: 145 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4258 gene can be obtained by the method described in Goff LA et al., 2009, PLoS One, vol. 4, e7192.
- hsa-miR-4258 “hsa-mir-4258” (miRBase Accession No. MI0015857, SEQ ID NO: 296) having a hairpin-like structure as a precursor is known.
- hsa-miR-4484 gene or “hsa-miR-4484” refers to the hsa-miR-4484 gene (miRBase Accession No. MIMAT0019018) described in SEQ ID NO: 146 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4484 gene can be obtained by the method described in Jim DD et al., 2010, Blood, 116, e118-e127.
- hsa-miR-4484 is known as “hsa-mir-4484” (miRBase Accession No. MI0016845, SEQ ID NO: 297), which has a hairpin-like structure as a precursor.
- hsa-miR-4648 gene or “hsa-miR-4648” refers to the hsa-miR-4648 gene (miRBase Accession No. MIMAT0019710) described in SEQ ID NO: 147 or other biological species. Includes homologs or orthologs.
- the hsa-miR-4648 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, 71, p78-86.
- “Hsa-miR-4648” is known as “hsa-mir-4648” (miRBase Accession No. MI0017275, SEQ ID NO: 298) having a hairpin-like structure as a precursor.
- hsa-miR-6780b-5p gene or “hsa-miR-6780b-5p” refers to the hsa-miR-6780b-5p gene described in SEQ ID NO: 148 (miRBase Accession No. MIMAT0027572) and other species homologs or orthologs.
- the hsa-miR-6780b-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p1634-1645.
- “hsa-miR-6780b-5p” is known as “hsa-mir-6780b” (miRBase Accession No. MI0022681, SEQ ID NO: 299) having a hairpin-like structure as a precursor.
- hsa-miR-4516 gene or “hsa-miR-4516” refers to the hsa-miR-4516 gene (miRBase Accession No. MIMAT0019053) described in SEQ ID NO: 466 or other biological species. Homologs or orthologs are included.
- the hsa-miR-4516 gene is disclosed in Jim DD et al., 2010, Blood. 116, e118-e127.
- hsa-mir-4516 (miRBase Accession No. MI0016882, SEQ ID NO: 479) having a hairpin-like structure as a precursor is known.
- hsa-miR-4649-5p gene or “hsa-miR-4649-5p” refers to the hsa-miR-4649-5p gene described in SEQ ID NO: 467 (miRBase Accession No. MIMAT0019711) and other species homologs or orthologs.
- the hsa-miR-4649-5p gene is disclosed in Persson H et al., 2011, Cancer Res. 71, p. 78-86.
- “hsa-miR-4649-5p” is known as “hsa-mir-4649” (miRBase Accession No. MI0017276, SEQ ID NO: 480) having a hairpin-like structure as a precursor.
- hsa-miR-760 gene or “hsa-miR-760” refers to the hsa-miR-760 gene (miRBase Accession No. MIMAT0004957) described in SEQ ID NO: 468 or other biological species. Homologs or orthologs are included. The hsa-miR-760 gene was obtained from Berezikov E et al., 2006, Genome Res. 16, p. 289-1298. As for “hsa-miR-760”, “hsa-mir-760” (miRBase Accession No. MI0005567, SEQ ID NO: 481) having a hairpin-like structure as a precursor is known.
- hsa-miR-3162-5p gene or “hsa-miR-3162-5p” refers to the hsa-miR-3162-5p gene described in SEQ ID NO: 469 (miRBase Accession No. MIMAT0015036) and other species homologs or orthologs.
- the hsa-miR-3162-5p gene was obtained from Stark MS et al., 2010, PLoS One. No. 5, vol. E9685.
- “hsa-miR-3162-5p” “hsa-mir-3162” (miRBase Accession No. MI0014192, SEQ ID NO: 482) having a hairpin-like structure as a precursor is known.
- hsa-miR-3178 gene or “hsa-miR-3178” refers to the hsa-miR-3178 gene (miRBase Accession No. MIMAT0015055) described in SEQ ID NO: 470 or other species. Homologs or orthologs are included. The hsa-miR-3178 gene was obtained from Stark MS et al., 2010, PLoS One. No. 5, vol. E9685. As for “hsa-miR-3178”, “hsa-mir-3178” (miRBase Accession No. MI0014212, SEQ ID NO: 483), which has a hairpin-like structure as a precursor, is known.
- hsa-miR-940 gene or “hsa-miR-940” refers to the hsa-miR-940 gene (miRBase Accession No. MIMAT0004983) described in SEQ ID NO: 471 or other biological species. Homologs or orthologs are included.
- the hsa-miR-940 gene is disclosed in Lui WO et al., 2007, Cancer Res. 67, p6031-6043.
- hsa-miR-940 “hsa-mir-940” (miRBase Accession No. MI0005762, SEQ ID NO: 484) having a hairpin-like structure as a precursor is known.
- hsa-miR-4271 gene or “hsa-miR-4271” refers to the hsa-miR-4271 gene (miRBase Accession No. MIMAT0016901) described in SEQ ID NO: 472 or other biological species. Homologs or orthologs are included. The hsa-miR-4271 gene was found in Goff LA et al., 2009, PLoS One. 4, Volume 4, e7192. In addition, “hsa-miR-4271” is known as “hsa-mir-4271” (miRBase Accession No. MI0015879, SEQ ID NO: 485), which has a hairpin-like structure as a precursor.
- hsa-miR-6769b-5p gene or “hsa-miR-6769b-5p” refers to the hsa-miR-6769b-5p gene (miRBase Accession No. MIMAT0027620) and other species homologues or orthologues are included.
- the hsa-miR-6769b-5p gene is described in Ladewig E et al., 2012, Genome Res. , Vol. 22, p1634-645.
- “hsa-miR-6769b-5p” is known as “hsa-mir-6769b” (miRBase Accession No. MI0022706, SEQ ID NO: 486), which has a hairpin-like structure as a precursor.
- hsa-miR-4508 gene or “hsa-miR-4508” refers to the hsa-miR-4508 gene (miRBase Accession No. MIMAT0019045) described in SEQ ID NO: 474 and other biological species. Homologs or orthologs are included.
- the hsa-miR-4508 gene is described in Jim DD et al., 2010, Blood. 116, e118-e127.
- hsa-mir-4508 (miRBase Accession No. MI0016872, SEQ ID NO: 487) having a hairpin-like structure as a precursor is known.
- hsa-miR-6826-5p gene or “hsa-miR-6826-5p” refers to the hsa-miR-6826-5p gene described in SEQ ID NO: 475 (miRBase Accession No. MIMAT0027552) and other species homologs or orthologs.
- the hsa-miR-6826-5p gene is disclosed in Ladewig E et al., 2012, Genome Res. , Vol. 22, p1634-645.
- hsa-miR-6826-5p “hsa-mir-6826” (miRBase Accession No. MI0022671, SEQ ID NO: 488) having a hairpin-like structure as a precursor is known.
- hsa-miR-6757-5p gene or “hsa-miR-6757-5p” refers to the hsa-miR-6757-5p gene described in SEQ ID NO: 476 (miRBase Accession No. MIMAT0027414) and other species homologues or orthologues are included.
- the hsa-miR-6757-5p gene is described in Ladewig E et al., 2012, Genome Res. , Vol. 22, p1634-645.
- hsa-miR-6757-5p “hsa-mir-6757” (miRBase Accession No. MI0022602, SEQ ID NO: 489) having a hairpin-like structure as a precursor is known.
- hsa-miR-3131 gene or “hsa-miR-3131” refers to the hsa-miR-3131 gene (miRBase Accession No. MIMAT0014996) described in SEQ ID NO: 477 or other biological species. Homologs or orthologs are included. The hsa-miR-3131 gene was obtained from Stark MS et al., 2010, PLoS One. No. 5, vol. E9685. Also, “hsa-miR-3131” is known as “hsa-mir-3131” (miRBase Accession No. MI0014151, SEQ ID NO: 490) having a hairpin-like structure as a precursor.
- hsa-miR-1343-3p gene or “hsa-miR-1343-3p” refers to the hsa-miR-1343-3p gene described in SEQ ID NO: 478 (miRBase Accession No. MIMAT0019776) and other species homologs or orthologs.
- the hsa-miR-1343-3p gene is disclosed in Persson H et al., 2011, Cancer Res. 71, p. 78-86.
- hsa-miR-1343-3p “hsa-mir-1343” (miRBase Accession No. MI0017320, SEQ ID NO: 491) having a hairpin-like structure as a precursor is known.
- miRNA when mature miRNA is cleaved as a mature miRNA from an RNA precursor having a hairpin-like structure, one to several bases before or after the sequence may be cleaved or long, or base substitution may occur. And is referred to as isomiR (Morin RD. Et al., 2008, Genome Research, Vol. 18, p. 610-621).
- miRBBase Release 20 in addition to the nucleotide sequence represented by any of SEQ ID NOs: 1 to 148, 466 to 478, the nucleotide sequence represented by any of SEQ ID NOs: 300 to 465 and 492 to 509, which are called numerous isomiRs, Variants and fragments are also shown. These variants can also be obtained as miRNA having the base sequence represented by any of SEQ ID NOs: 1 to 148 and 466 to 478.
- SEQ ID NOs: 1, 3, 4, 6, 14, 16, 17, 18, 22, 23, 24, 25, 30, 31, 34 registered in miRBBase 35, 37, 42, 43, 44, 47, 48, 49, 50, 51, 52, 55, 57, 59, 61, 62, 66, 67, 69, 70, 72, 73, 75, 77, 79, 80, 82, 83, 84, 85, 86, 89, 90, 92, 94, 96, 99, 101, 102, 103, 104, 106, 107, 109, 110, 111, 112, 113, 115, 116, Polynucleotides 120, 121, 122, 124, 130, 131, 132, 133, 136, 137, 138, 139, 140, 141, 142, 144, 146, and 147, which are numerous isomiRs.
- examples of the polynucleotide containing the base sequence represented by any of SEQ ID NOs: 1 to 148, 466 to 478 are represented by any of the precursors SEQ ID NOs: 149 to 299 and 479 to 491, respectively.
- a polynucleotide is mentioned.
- nucleic acid probe or primer used in the present invention binds to a specific target nucleic acid and cannot substantially bind to another nucleic acid.
- the present invention makes it possible to detect biliary tract cancer easily and with high accuracy. For example, it is possible to easily detect whether or not a patient has a biliary tract cancer using as an index several measurement values of miRNA in blood, serum and / or plasma of a patient that can be collected minimally invasively.
- This figure shows hsa-miR-4665-5p represented by SEQ ID NO: 51 generated from hsa-mir-4665 represented by SEQ ID NO: 201 which is a precursor, and hsa-miR represented by SEQ ID NO: 91.
- the relationship of the base sequence of ⁇ 4665-3p is shown.
- the horizontal line in the figure shows the threshold (5.69) for discriminating both groups, optimized by Fisher's discriminant analysis.
- 67 biliary tract cancer patients 93 healthy subjects, 35 colorectal cancer patients, 37 stomach cancer patients, 32 esophageal cancer patients, 38 liver cancer patients, and benign pancreaticobiliary tract benign selected as a learning sample group 13 patients with disease, hsa-miR-6075 (SEQ ID NO: 15), hsa-miR-6636-3p (SEQ ID NO: 12), hsa-miR-6799-5p (SEQ ID NO: 29), hsa-miR-125a-3p ( A discriminant is created from the measured value of SEQ ID NO: 1 using Fisher's discriminant analysis ( ⁇ 1.25 ⁇ hsa-miR-6075 ⁇ 1.06 ⁇ hsa-miR-6836-3p + 0.53 ⁇ hsa-miR ⁇ ) 6799-5p + 0.18 ⁇ hsa-miR-125a-3p + 15.41), where the discriminant score obtained from the discriminant is represented on the vertical
- the dotted line in the figure indicates a discrimination boundary for discriminating both groups having a discrimination score of 0.
- the discrimination score obtained from the discriminant created in the learning sample group is represented on the vertical axis, and the sample group is represented on the horizontal axis.
- the dotted line in the figure indicates a discrimination boundary for discriminating both groups having a discrimination score of 0.
- Target nucleic acid for biliary tract cancer The biliary tract for detecting the presence and / or absence of biliary tract cancer or biliary tract cancer cells using the nucleic acid probe or primer for detection of biliary tract cancer as defined above of the present invention is provided.
- Major target nucleic acids as cancer markers include hsa-miR-125a-3p, hsa-miR-6893-5p, hsa-miR-204-3p, hsa-miR-4476, hsa-miR-4294, hsa-miR -150-3p, hsa-miR-6729-5p, hsa-miR-7641, hsa-miR-6765-3p, hsa-miR-6820-5p, hsa-miR-575, hsa-miR-6683-3p, hsa -MiR-1469, hsa-miR-663a, hsa-miR-6075, hsa-miR-4634, hsa-m iR-423-5p, hsa-miR-4454, hsa-miR-7109-5p,
- biliary tract cancer markers that can be combined with these miRNAs are: hsa-miR-6808-5p, hsa-miR-6774-5p, hsa-miR-4656, hsa-miR-6806-5p, hsa- miR-1233-5p, hsa-miR-328-5p, hsa-miR-4675, hsa-miR-2110, hsa-miR-6076, hsa-miR-3619-3p, hsa-miR-92a-2-5p, hsa-miR-128-1-5p, hsa-miR-638, hsa-miR-2861, hsa-miR-371a-5p, hsa-miR-211-3p, hsa-miR-1273g-3p, hsa-miR- 1203, hsa
- the miRNA includes, for example, human genes containing the nucleotide sequence represented by any of SEQ ID NOs: 1 to 148 and 466 to 478 (ie, hsa-miR-125a-3p and hsa-miR-6893-5p, respectively).
- a preferred target nucleic acid is a human gene comprising the nucleotide sequence represented by any of SEQ ID NOs: 1 to 509, or a transcription product thereof, more preferably the transcription product, ie, miRNA, a pri-miRNA that is a precursor RNA thereof. Or pre-miRNA.
- the first target gene is the hsa-miR-125a-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the second target gene is the hsa-miR-6893-5p gene, their homologues, their transcripts, or their mutants or derivatives. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the third target gene is the hsa-miR-204-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the fourth target gene is the hsa-miR-4476 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the fifth target gene is the hsa-miR-4294 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the sixth target gene is the hsa-miR-150-3p gene, their homologues, their transcripts, or their mutants or derivatives. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the seventh target gene is the hsa-miR-6729-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the eighth target gene is the hsa-miR-7641 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the ninth target gene is the hsa-miR-6765-3p gene, their homologues, their transcripts, or their mutants or derivatives. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the tenth target gene is the hsa-miR-6820-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the eleventh target gene is the hsa-miR-575 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the twelfth target gene is the hsa-miR-6836-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- hsa-miR-6836-3p the gene or its transcript can serve as a marker for biliary tract cancer.
- the thirteenth target gene is the hsa-miR-1469 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the fourteenth target gene is an hsa-miR-663a gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- hsa-miR-663a gene a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the fifteenth target gene is the hsa-miR-6075 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 16th target gene is the hsa-miR-4634 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 17th target gene is the hsa-miR-423-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 18th target gene is the hsa-miR-4454 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the nineteenth target gene is the hsa-miR-7109-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- this gene or its transcript can serve as a marker for biliary tract cancer.
- the twentieth target gene is the hsa-miR-6789-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- this gene or its transcript can serve as a marker for biliary tract cancer.
- the 21st target gene is the hsa-miR-6877-5p gene, their homologues, their transcripts, or their mutants or derivatives. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 22nd target gene is the hsa-miR-4792 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 23rd target gene is the hsa-miR-4530 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 24th target gene is the hsa-miR-7975 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 25th target gene is the hsa-miR-6724-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the twenty-sixth target gene is the hsa-miR-8073 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- this gene or its transcript can serve as a marker for biliary tract cancer.
- the 27th target gene is the hsa-miR-7777 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 28th target gene is the hsa-miR-1231 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 29th target gene is the hsa-miR-6799-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 30th target gene is the hsa-miR-615-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the thirty-first target gene is the hsa-miR-4450 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- this gene or its transcript can serve as a marker for biliary tract cancer.
- the thirty-second target gene is the hsa-miR-6726-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- this gene or its transcript can serve as a marker for biliary tract cancer.
- the 33rd target gene is the hsa-miR-6875-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 34th target gene is the hsa-miR-4734 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 35th target gene is an hsa-miR-16-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the thirty-sixth target gene is an hsa-miR-602 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- hsa-miR-602 a homologue thereof
- a transcription product thereof or a mutant or derivative thereof.
- the 37th target gene is the hsa-miR-4651 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 38th target gene is the hsa-miR-8069 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 39th target gene is an hsa-miR-1238-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 40th target gene is the hsa-miR-6880-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 41st target gene is the hsa-miR-8072 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the forty-second target gene is the hsa-miR-4723-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 43rd target gene is the hsa-miR-4732-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 44th target gene is the hsa-miR-6125 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 45th target gene is the hsa-miR-6090 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 46th target gene is the hsa-miR-7114-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 47th target gene is the hsa-miR-564 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 48th target gene is the hsa-miR-451a gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 49th target gene is the hsa-miR-3135b gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 50th target gene is the hsa-miR-4497 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 51st target gene is the hsa-miR-4665-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 52nd target gene is the hsa-miR-3622a-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 53rd target gene is the hsa-miR-6850-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 54th target gene is the hsa-miR-6821-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 55th target gene is the hsa-miR-5100 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 56th target gene is the hsa-miR-6872-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 57th target gene is the hsa-miR-4433-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 58th target gene is the hsa-miR-1227-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- this gene or its transcript can serve as a marker for biliary tract cancer.
- the 59th target gene is the hsa-miR-3188 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 60th target gene is the hsa-miR-7704 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 61st target gene is the hsa-miR-3185 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 62nd target gene is the hsa-miR-1908-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 63rd target gene is the hsa-miR-6781-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 64th target gene is the hsa-miR-6805-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 65th target gene is the hsa-miR-8089 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 66th target gene is an hsa-miR-665 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 67th target gene is the hsa-miR-4486 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 68th target gene is the hsa-miR-6722-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 69th target gene is the hsa-miR-1260a gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 70th target gene is the hsa-miR-4707-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 71st target gene is the hsa-miR-6741-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 72nd target gene is the hsa-miR-1260b gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 73rd target gene is the hsa-miR-1246 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 74th target gene is the hsa-miR-6845-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 75th target gene is the hsa-miR-4638-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 76th target gene is the hsa-miR-6085 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 77th target gene is the hsa-miR-1228-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 78th target gene is the hsa-miR-4534 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 79th target gene is the hsa-miR-5585-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 80th target gene is the hsa-miR-4741 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 81st target gene is the hsa-miR-4433b-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 82nd target gene is the hsa-miR-197-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 83rd target gene is the hsa-miR-718 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 84th target gene is the hsa-miR-4513 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 85th target gene is the hsa-miR-4446-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 86th target gene is the hsa-miR-619-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 87th target gene is the hsa-miR-6816-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 88th target gene is the hsa-miR-6778-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 89th target gene is an hsa-miR-24-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 90th target gene is the hsa-miR-1915-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 91st target gene is the hsa-miR-4665-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 92nd target gene is the hsa-miR-4449 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 93rd target gene is the hsa-miR-6889-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 94th target gene is the hsa-miR-486-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 95th target gene is the hsa-miR-7113-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 96th target gene is the hsa-miR-642a-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 97th target gene is the hsa-miR-7847-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 98th target gene is the hsa-miR-6768-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 99th target gene is an hsa-miR-1290 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 100th target gene is the hsa-miR-7108-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 101st target gene is the hsa-miR-92b-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 102nd target gene is the hsa-miR-663b gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 103rd target gene is the hsa-miR-3940-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 104th target gene is the hsa-miR-4467 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 105th target gene is the hsa-miR-6858-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 106th target gene is the hsa-miR-4417 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 107th target gene is the hsa-miR-3665 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- hsa-miR-3665 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 108th target gene is the hsa-miR-4736 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 109th target gene is the hsa-miR-4687-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 110th target gene is the hsa-miR-1908-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 111th target gene is the hsa-miR-5195-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 112th target gene is the hsa-miR-4286 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 113th target gene is the hsa-miR-3679-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 114th target gene is the hsa-miR-6791-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 115th target gene is the hsa-miR-1220 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 116th target gene is the hsa-miR-3656 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 117th target gene is the hsa-miR-4746-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- hsa-miR-4746-3p the gene or its transcript can serve as a marker for biliary tract cancer.
- the 118th target gene is the hsa-miR-3184-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 119th target gene is the hsa-miR-3937 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 120th target gene is the hsa-miR-6515-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 121st target gene is the hsa-miR-6132 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 122nd target gene is the hsa-miR-187-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 123rd target gene is the hsa-miR-7111-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 124th target gene is the hsa-miR-5787 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 125th target gene is an hsa-miR-6679-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- hsa-miR-6679-5p gene a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 126th target gene is the hsa-miR-6808-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 127th target gene is the hsa-miR-6774-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 128th target gene is the hsa-miR-4656 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 129th target gene is the hsa-miR-6806-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 130th target gene is the hsa-miR-1233-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 131st target gene is an hsa-miR-328-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 132nd target gene is the hsa-miR-4675 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 133rd target gene is the hsa-miR-2110 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 134th target gene is the hsa-miR-6076 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 135th target gene is the hsa-miR-3619-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 136th target gene is the hsa-miR-92a-2-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 137th target gene is the hsa-miR-128-1-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 138th target gene is the hsa-miR-638 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- a homologue thereof a transcription product thereof, or a mutant or derivative thereof.
- the 139th target gene is the hsa-miR-2861 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 140th target gene is the hsa-miR-371a-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 141st target gene is the hsa-miR-211-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 142nd target gene is the hsa-miR-1273g-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 143rd target gene is the hsa-miR-1203 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 144th target gene is the hsa-miR-122-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 145th target gene is the hsa-miR-4258 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 146th target gene is the hsa-miR-4484 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 147th target gene is the hsa-miR-4648 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 148th target gene is the hsa-miR-6780b-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof. Until now, there has been no report that changes in the expression of this gene or its transcript can serve as a marker for biliary tract cancer.
- the 149th target gene is the hsa-miR-4516 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 150th target gene is the hsa-miR-4649-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 151st target gene is an hsa-miR-760 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 152th target gene is the hsa-miR-3162-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 153rd target gene is the hsa-miR-3178 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 154th target gene is the hsa-miR-940 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 155th target gene is the hsa-miR-4271 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 156th target gene is the hsa-miR-6769b-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 157th target gene is the hsa-miR-4508 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 158th target gene is the hsa-miR-6826-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 159th target gene is the hsa-miR-6757-5p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 160th target gene is the hsa-miR-3131 gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- the 161st target gene is the hsa-miR-1343-3p gene, a homologue thereof, a transcription product thereof, or a mutant or derivative thereof.
- nucleic acid probe or primer for detection of biliary tract cancer a nucleic acid capable of specifically binding to the target nucleic acid as the above biliary tract cancer marker, a nucleic acid for detecting or diagnosing biliary tract cancer, for example, a nucleic acid It can be used as a probe or primer.
- the nucleic acid probe or primer that can be used for detecting or diagnosing biliary tract cancer is a target nucleic acid as the above-mentioned biliary tract cancer marker, for example, human-derived hsa-miR.
- the above target nucleic acids may increase or decrease in the expression level of the target nucleic acid depending on the type of the target nucleic acid in a subject suffering from biliary tract cancer compared to a healthy subject (hereinafter referred to as “the target nucleic acid”). , Referred to as “increase / decrease”). Therefore, the nucleic acid of the present invention measures the expression level of the target nucleic acid in a body fluid derived from a subject (eg, human) suspected of having biliary tract cancer and a body fluid derived from a healthy body, and compares them to determine the biliary tract. It can be used effectively to detect cancer.
- a subject eg, human
- the nucleic acid of the present invention comprises a body fluid derived from a subject (eg, human) suspected of having biliary tract cancer, a colorectal cancer patient, a stomach cancer patient, an esophageal cancer patient, a liver cancer patient, and a pancreaticobiliary benign disease patient. It can be used effectively to measure biliary tract cancer specifically from other cancers or benign diseases by measuring the expression level of the target nucleic acid from the derived body fluid and comparing them.
- a subject eg, human
- the nucleic acid probe or primer that can be used in the present invention is specific to a polynucleotide comprising a base sequence represented by at least one of SEQ ID NOs: 1-125 (preferably SEQ ID NOs: 1, 2, 4-125) and 466-478. Or a primer for amplifying a polynucleotide comprising a nucleotide sequence represented by at least one of SEQ ID NOs: 1-125, 466-478.
- the nucleic acid probe or primer that can be used in the present invention is further a nucleic acid probe that can specifically bind to a polynucleotide consisting of the base sequence represented by at least one of SEQ ID NOs: 126 to 148, or of SEQ ID NOs: 126 to 148 Primers for amplifying a polynucleotide comprising the base sequence represented by at least one can be included.
- the nucleic acid probe or primer includes a polynucleotide group including a base sequence represented by any of SEQ ID NOs: 1 to 509, or a base sequence in which u is t in the base sequence, and a complementary sequence thereof
- nucleic acid probes or primers that can be used in the present invention are one or more polynucleotides selected from the group consisting of the following polynucleotides (a) to (e).
- A a polynucleotide comprising the base sequence represented by any of SEQ ID NOs: 1-125, 466-478, or a base sequence in which u is t in the base sequence, a variant thereof, a derivative thereof, or 15 or more A fragment thereof containing a continuous base of
- B a polynucleotide comprising the base sequence represented by any of SEQ ID NOs: 1-125, 466-478
- C a polynucleotide comprising a base sequence complementary to the base sequence represented by any of SEQ ID NOs: 1-125, 466-478, or a base sequence in which u is t in the base sequence, variants thereof, A derivative thereof, or a fragment thereof comprising 15 or more consecutive bases
- D a polynucleotide
- the nucleic acid probe or primer that can be used in the present invention further includes the following (f) to (j And a polynucleotide selected from the group consisting of the polynucleotides shown in FIG.
- the fragment containing 15 or more consecutive bases refers to, for example, 15 to less than the total number of bases in the sequence, 17 to less than the total number of bases in the sequence, 19 To less than the total number of bases in the sequence, etc., but not limited thereto.
- Any of the above polynucleotides or fragments thereof used in the present invention may be DNA or RNA.
- the above-mentioned polynucleotide that can be used in the present invention can be prepared using a general technique such as a DNA recombination technique, a PCR method, or a method using a DNA / RNA automatic synthesizer.
- DNA recombination techniques and PCR methods include, for example, Ausubel et al., Current Protocols in Molecular Biology, John Willy & Sons, US (1993); Sambrook et al., Molecular Cloning A Laboratory A Laboratory. The techniques described can be used.
- Such a nucleic acid probe or primer can be chemically synthesized using an automatic DNA synthesizer.
- the phosphoramidite method is used for this synthesis, and single-stranded DNA of up to about 100 bases can be automatically synthesized by this method.
- Automatic DNA synthesizers are commercially available from, for example, Polygen, ABI, Applied BioSystems, and the like.
- the polynucleotide of the present invention can also be prepared by a cDNA cloning method.
- a cDNA cloning method for example, microRNA Cloning Kit Wako can be used as the cDNA cloning technique.
- the nucleic acid probe and primer sequences for detecting a polynucleotide comprising the nucleotide sequence represented by any of SEQ ID NOs: 1 to 148 and 466 to 478 are present in vivo as miRNA or a precursor thereof.
- the base sequences represented by SEQ ID NO: 51 and SEQ ID NO: 91 are generated from the precursor represented by SEQ ID NO: 201, and this precursor has a hairpin-like structure as shown in FIG.
- the base sequences represented by SEQ ID NO: 51 and SEQ ID NO: 91 have mismatched sequences. For this reason, a completely complementary base sequence to the base sequence represented by SEQ ID NO: 51 or SEQ ID NO: 91 is not naturally generated in vivo.
- the nucleic acid probe and primer for detecting the base sequence represented by any of SEQ ID NOs: 1 to 148 and 466 to 478 have an artificial base sequence that does not exist in the living body.
- Biliary tract cancer detection kit or device The present invention also provides a polynucleotide that can be used as a nucleic acid probe or primer in the present invention for measuring a target nucleic acid that is a biliary tract cancer marker (including variants, fragments, A kit or device for detecting biliary tract cancer is provided that includes one or more of the following.
- the target nucleic acid that is a biliary tract cancer marker in the present invention is preferably selected from the following group 1: miR-125a-3p, miR-6893-5p, miR-204-3p, miR-4476, miR-4294, miR-150-3p, miR-6729-5p, miR-7641, miR-6765-3p, miR- 6820-5p, miR-575, miR-6683-3p, miR-1469, miR-663a, miR-6075, miR-4634, miR-423-5p, miR-4454, miR-7109-5p, miR-6789- 5p, miR-6877-5p, miR-4792, miR-4530, miR-7975, miR-6724-5p, miR-8073, miR-7777, miR-1231, miR-6799-5p, miR-615-5p, miR-4450, miR-6726-5p, mi -6875-5p, miR-47
- the additional target nucleic acid that can optionally be used for the measurement is preferably selected from the following group 2: miR-6808-5p, miR-6774-5p, miR-4656, miR-6806-5p, miR-1233 -5p, miR-328-5p, miR-4675, miR-2110, miR-6076, miR-3619-3p, miR-92a-2-5p, miR-128-1-5p, miR-638, miR-2861 MiR-371a-5p, miR-211-3p, miR-1273g-3p, miR-1203, miR-122-5p, miR-4258, miR-4484, miR-4680 and miR-6780b-5p.
- the kit or device of the present invention is a nucleic acid that can specifically bind to a target nucleic acid that is the above-mentioned biliary tract cancer marker, preferably the nucleic acid probe or primer described in 2 above, specifically the polynuclear acid described in 2 above. It includes one or more polynucleotides selected from nucleotides or variants thereof.
- the kit or device of the present invention includes a base sequence represented by any of SEQ ID NOs: 1-125 and 466-478, or a base sequence in which u is t in the base sequence (or A polynucleotide comprising (or consisting of) a complementary sequence thereof, a polynucleotide that hybridizes with these polynucleotides under stringent conditions, or 15 or more consecutive bases of those polynucleotide sequences. At least one or more mutants or fragments.
- the kit or device of the present invention further comprises a polynucleotide comprising (or consisting of) the nucleotide sequence represented by any of SEQ ID NOs: 126 to 148, or a nucleotide sequence in which u is t in the nucleotide sequence, and its complement
- a polynucleotide comprising (or consisting of) a target sequence, a polynucleotide that hybridizes with these polynucleotides under stringent conditions, or a variant or fragment comprising 15 or more consecutive bases of those polynucleotide sequences, One or more can be included.
- the fragment that can be included in the kit or device of the present invention is, for example, one or more, preferably two or more polynucleotides selected from the group consisting of the following (1) to (2).
- (1) A polynucleotide comprising 15 or more consecutive bases in a base sequence represented by any one of SEQ ID NOS: 1-125, 466-478 or a complementary sequence thereof wherein u is t.
- (2) A polynucleotide comprising 15 or more consecutive bases in the base sequence represented by any one of SEQ ID NOs: 126 to 148 wherein u is t or a complementary sequence thereof.
- the polynucleotide is a polynucleotide comprising a base sequence represented by any one of SEQ ID NOs: 1-125, 466-478, or a base sequence in which u is t in the base sequence, its complementary A polynucleotide comprising a sequence, a polynucleotide that hybridizes with these polynucleotides under stringent conditions, or a variant containing 15 or more, preferably 17 or more, more preferably 19 or more consecutive bases thereof.
- the polynucleotide is a polynucleotide comprising the nucleotide sequence represented by any of SEQ ID NOs: 126 to 148, or a nucleotide sequence in which u is t in the nucleotide sequence, or a complementary sequence thereof. Or a polynucleotide that hybridizes with these polynucleotides under stringent conditions, or a variant containing 15 or more, preferably 17 or more, more preferably 19 or more consecutive bases thereof.
- the fragment may be a polynucleotide comprising 15 or more, preferably 17 or more, more preferably 19 or more consecutive bases.
- the size of a polynucleotide fragment is, for example, 15 to less than the total number of bases in the sequence, 17 to less than the total number of bases in the sequence, and 19 to less than the total number of bases in the sequence.
- the number of bases in the range is, for example, 15 to less than the total number of bases in the sequence, 17 to less than the total number of bases in the sequence, and 19 to less than the total number of bases in the sequence. The number of bases in the range.
- kits or device of the present invention include SEQ ID NOs: 1 to 148 and 466 corresponding to miRNA markers in Table 1 described later in Table 1.
- any combination of the above-mentioned polynucleotides comprising the base sequence represented by or a complementary sequence thereof can be mentioned, but these are merely examples, and all other various possible combinations are included in the present invention. Shall be included.
- the above-mentioned combination constituting the kit or device for discriminating biliary tract cancers from healthy bodies includes two of the above-mentioned polynucleotides comprising the base sequences represented by the sequence numbers shown in Table 1. It is desirable to combine them as described above. Usually, sufficient performance can be obtained with two combinations.
- a combination of two polynucleotides comprising a base sequence for distinguishing biliary tract cancer from a healthy body or a complementary sequence thereof, the above-described base sequences comprising the base sequences represented by SEQ ID NOs: 1 to 148 and 466 to 478 Of the two combinations selected from these polynucleotides, a combination comprising at least one or more of polynucleotides consisting of the nucleotide sequences represented by SEQ ID NOs: 1-125 and 466-478 that are newly found is preferable.
- examples of combinations of polynucleotides having cancer type specificity that can distinguish biliary tract cancer from not only healthy bodies but also other cancers include, for example, SEQ ID NOs: 1, 4, 5, 11, 12, 15, 23, 29. , 39, 40, 54, 76, 79, 91, 103, 115, 121, 134, 143, 466, 469, 472, 473, and 474, or a group consisting of polynucleotides comprising a complementary sequence thereof (Hereinafter, this group is referred to as “cancer type-specific polynucleotide group 1”) and a plurality of combinations of at least one polynucleotide selected from the other polynucleotides of SEQ ID NO. Are preferable.
- a combination of polynucleotides having cancer type specificity that can distinguish biliary tract cancer from not only healthy bodies but also other cancers a plurality of polynucleotides selected from cancer type specific polynucleotide group 1 are used. A combination is more preferred.
- a plurality of polynucleotides selected from cancer type specific polynucleotide group 1 are used.
- a group consisting of a nucleotide sequence represented by SEQ ID NOs: 4, 5, 12, 15 and 40 or a polynucleotide comprising a complementary sequence thereof, which is included in the cancer type-specific polynucleotide group 1 hereinafter referred to as this book
- a combination comprising at least one polynucleotide selected from “group of cancer type-specific polynucleotide group 2”) is more preferable.
- the number of combinations of the above-mentioned cancer type-specific polynucleotides is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. However, it is more preferable that the number of combinations is four or more. Usually, a sufficient performance can be obtained with four combinations.
- a polynucleotide comprising the nucleotide sequence represented by SEQ ID NO: 4 or a complementary sequence thereof and three polynucleotides selected from the cancer-species specific polynucleotide group 1 are represented by SEQ ID NO: Examples of the combination with a polynucleotide comprising a base sequence or a complementary sequence thereof are given below.
- a polynucleotide comprising the nucleotide sequence represented by SEQ ID NO: 5 or its complementary sequence, and three polynucleotides selected from the cancer-type specific polynucleotide group 1 are represented by SEQ ID NO: Examples of the combination with a polynucleotide comprising a base sequence or a complementary sequence thereof are given below.
- a polynucleotide comprising the nucleotide sequence represented by SEQ ID NO: 12 or a complementary sequence thereof and three polynucleotides selected from the cancer species-specific polynucleotide group 1 are represented by SEQ ID NOs: Examples of the combination with a polynucleotide comprising a base sequence or a complementary sequence thereof are given below.
- a polynucleotide comprising the nucleotide sequence represented by SEQ ID NO: 15 or its complementary sequence and three polynucleotides selected from the cancer type-specific polynucleotide group 1 are represented by SEQ ID NOs: Examples of the combination with a polynucleotide comprising a base sequence or a complementary sequence thereof are given below.
- polynucleotide selected from, but not limited to, the polynucleotide consisting of the base sequence represented by SEQ ID NO: 40 or its complementary sequence and the cancer type-specific polynucleotide group 1.
- SEQ ID NO: 40 the polynucleotide consisting of the base sequence represented by SEQ ID NO: 40 or its complementary sequence
- cancer type-specific polynucleotide group 1 the cancer type-specific polynucleotide group 1. Examples of the combination with a polynucleotide comprising a base sequence or a complementary sequence thereof are given below.
- the kit or device of the present invention includes known polynucleotides that enable detection of biliary tract cancer in addition to the polynucleotides of the present invention described above (which may include mutants, fragments or derivatives). Nucleotides or polynucleotides that may be found in the future can also be included.
- the kit of the present invention includes CEA, CA19-9, Span-1, DUPAN-2, CA50, CA195, IL-6, CA242, TAG-72, urine in addition to the polynucleotide of the present invention described above.
- Antibodies for measuring known biliary tract cancer test markers such as fucose, POA, TPS and the like can also be included.
- the polynucleotides contained in the kit of the present invention can be individually or arbitrarily combined and packaged in different containers.
- the kit of the present invention can include a kit for extracting nucleic acid (for example, total RNA) from body fluids, cells or tissues, a fluorescent substance for labeling, an enzyme and medium for nucleic acid amplification, instructions for use, and the like.
- nucleic acid for example, total RNA
- the device of the present invention is a device for measuring a cancer marker in which a nucleic acid such as a polynucleotide according to the present invention described above is bound or attached to a solid phase, for example.
- a nucleic acid such as a polynucleotide according to the present invention described above is bound or attached to a solid phase
- the material of the solid phase are plastic, paper, glass, silicon, and the like. From the viewpoint of ease of processing, a preferable material of the solid phase is plastic.
- the shape of the solid phase is arbitrary, for example, a square shape, a round shape, a strip shape, a film shape and the like.
- Examples of the device of the present invention include a device for measurement by a hybridization technique, and specific examples include a blotting device, a nucleic acid array (for example, a microarray, a DNA chip, an RNA chip, etc.).
- the nucleic acid array technology uses a high-density dispenser called a spotter or arrayer on the surface of a solid phase that has been subjected to surface treatment such as introduction of functional groups such as L-lysine coat, amino group, and carboxyl group as necessary.
- the method of spotting nucleic acids the method of spraying nucleic acids onto a solid phase using an inkjet that ejects fine droplets from a nozzle with a piezoelectric element, the method of sequentially synthesizing nucleotides on a solid phase, etc.
- an array such as a chip is produced by binding or attaching the nucleic acids one by one, and the target nucleic acid is measured using hybridization using this array.
- the kit or device of the present invention comprises at least one, preferably at least two, more preferably at least three, most preferably at least five or all of miRNAs that are the above-mentioned group 1 biliary tract cancer markers.
- the kit or device of the present invention may further optionally include at least one, preferably at least two, more preferably at least three, and most preferably five of the above-mentioned group 2 biliary tract cancer markers miRNA. Nucleic acids that can specifically bind to each of all polynucleotides can be included.
- the kit or device of the present invention can be used for the detection of the following biliary tract cancers.
- Biliary tract cancer detection method The present invention further includes the above-mentioned 3.
- the above method of the present invention enables early diagnosis of cancer with minimal invasiveness, high sensitivity and specificity, thereby leading to early treatment and improvement of prognosis, as well as monitoring disease aversion and surgical Enables monitoring of the effectiveness of radiotherapeutic and chemotherapeutic treatments.
- RNA extraction reagent liquid sample kit (Toray Industries, Inc.)
- a general acidic phenol method (Acid Guanidinium-Phenol-Chloroform (AGPC) method) or Trizol (registered trademark) (Life Technologies)
- AGPC Acid Guanidinium-Phenol-Chloroform
- Trizol registered trademark
- kits such as miRNeasy (registered trademark) Mini Kit (Qiagen) can be used, but are not limited to these methods.
- the present invention also provides the use of the kit or device of the present invention for in vitro detection of an expression product of a miRNA gene derived from biliary tract cancer in a specimen derived from a subject.
- the kit or device is used as described above, and includes a single or any possible combination of polynucleotides that can be used in the present invention.
- the polynucleotide contained in the kit or device of the present invention can be used as a probe or primer.
- a primer Life Technologies' TaqMan (registered trademark) MicroRNA Assays, Qiagen's miScript PCR System, and the like can be used, but are not limited thereto.
- Polynucleotides contained in the kit or device of the present invention are quantitatively determined by hybridization techniques such as Northern blotting, Southern blotting, in situ hybridization, Northern hybridization, Southern hybridization, and quantitative RT-PCR.
- a known method for specifically detecting a specific gene such as an amplification technique, it can be used as a primer or a probe according to a conventional method.
- body fluid such as blood, serum, plasma, urine, etc. of the subject is collected according to the type of detection method used.
- total RNA prepared by the above-described method may be used, or various polynucleotides containing cDNA prepared based on the RNA may be used.
- the kit or device of the present invention is useful for diagnosis of biliary tract cancer or detection of the presence or absence of morbidity.
- the detection of biliary tract cancer using the kit or device is carried out by using a specimen such as blood, serum, plasma, urine or the like from a subject suspected of having biliary tract cancer.
- the detection can be performed in vitro by detecting the expression level of the gene detected by the nucleic acid probe or primer contained therein.
- the expression level of the target miRNA marker measured by a polynucleotide comprising a nucleotide sequence represented by one or more of SEQ ID NOs: 126 to 148 or a complementary sequence thereof (including variants, fragments or derivatives thereof)
- the subject is evaluated as having biliary tract cancer. can do.
- the method of the present invention can be combined with diagnostic imaging methods such as abdominal ultrasonography, CT scan, endoscopic retrograde cholangiopancreatography, and ultrasonography.
- the method of the present invention can specifically detect biliary tract cancer and can be substantially distinguished from cancers other than biliary tract cancer. Especially in the case of pancreatic cancer, it is possible to use a miRNA marker that is partly the same as that in biliary tract cancer. These cancers can be identified by combining with other diagnostic methods such as diagnostic imaging methods as described above.
- a method of detecting that a sample using a kit or device of the present invention does not contain an expression product of a gene derived from biliary tract cancer or an expression product of a gene derived from biliary tract cancer is contained in a subject.
- a body fluid such as blood, serum, plasma, urine is collected, and the expression level of the target gene contained therein is determined according to one or a plurality of polynucleotides (mutants, fragments or derivatives) selected from the polynucleotide group of the present invention. Evaluation of the presence or absence of biliary tract cancer, or detection of biliary tract cancer.
- a therapeutic agent when administered for the improvement of the disease, the presence or absence of improvement of the disease or the degree of improvement is evaluated or diagnosed. You can also.
- the method of the present invention includes, for example, the following steps (a), (b) and (c): (A) contacting a specimen from a subject with a polynucleotide in a kit or device of the present invention in vitro; (B) measuring the expression level of the target nucleic acid in the specimen using the polynucleotide as a nucleic acid probe or primer; (C) based on the result of (b), evaluating the presence or absence of biliary tract cancer (cells) in the subject, Can be included.
- the present invention relates to miR-125a-3p, miR-6893-5p, miR-204-3p, miR-4476, miR-4294, miR-150-3p, miR-6729-5p, miR-7641.
- evaluation is not an evaluation by a doctor but an evaluation support based on a result of an in vitro examination.
- miR-125a-3p is hsa-miR-125a-3p and miR-6893-5p is hsa-miR-6893-5p.
- MiR-204-3p is hsa-miR-204-3p
- miR-4476 is hsa-miR-4476
- miR-4294 is hsa-miR-4294
- miR-150-3p is hsa-miR.
- miR-6729-5p is hsa-miR-6729-5p
- miR-7641 is hsa-miR-7641
- miR-6765-3p is hsa-miR-6765-3p
- MiR-6820-5p is hsa-miR-6820-5p and miR-575 is hsa miR-575
- miR-6636-3p is hsa-miR-6636-3p
- miR-1469 is hsa-miR-1469
- miR-663a is hsa-miR-663a
- miR-6075 is hsa-miR-6075
- miR-4634 is hsa-miR-4634
- miR-423-5p is hsa-miR-423-5p
- miR-4454 is hsa-miR-4454
- miR- 7109-5p is hsa-miR-7109-5
- the nucleic acid is a polynucleotide shown in the following (a) to (e): (A) a polynucleotide comprising the base sequence represented by any of SEQ ID NOs: 1-125, 466-478, or a base sequence in which u is t in the base sequence, a variant thereof, a derivative thereof, or 15 or more A fragment thereof containing a continuous base of (B) a polynucleotide comprising the base sequence represented by any of SEQ ID NOs: 1-125, 466-478, (C) a polynucleotide comprising a base sequence complementary to the base sequence represented by any of SEQ ID NOs: 1-125, 466-478, or a base sequence in which u is t in the base sequence, variants thereof, A derivative thereof, or a fragment thereof comprising 15 or more consecutive bases, (D) a polynucleotide comprising a base sequence
- such a nucleic acid is specifically miR-6808-5p is hsa-miR-6808-5p, miR-6774-5p is hsa-miR-6774-5p, and miR- 4656 is hsa-miR-4656, miR-6806-5p is hsa-miR-6806-5p, miR-1233-5p is hsa-miR-1233-5p, and miR-328-5p is hsa- miR-328-5p, miR-4675 is hsa-miR-4675, miR-2110 is hsa-miR-2110, miR-6076 is hsa-miR-6076, and miR-3619-3p is hsa-miR-3619-3p, and miR-92a-2-5p is hsa-miR-9 a-2-5p, miR-128-1-5p is hsa-miR-128-1
- such a nucleic acid is a polynucleotide shown in the following (f) to (j): (F) a polynucleotide comprising the nucleotide sequence represented by any of SEQ ID NOs: 126 to 148, or a nucleotide sequence in which u is t in the nucleotide sequence, a variant thereof, a derivative thereof, or 15 or more consecutive bases Its fragments, including (G) a polynucleotide comprising the base sequence represented by any of SEQ ID NOs: 126 to 148, (H) a polynucleotide comprising a nucleotide sequence complementary to the nucleotide sequence represented by any of SEQ ID NOs: 126 to 148, or a nucleotide sequence in which u is t in the nucleotide sequence, a variant thereof, a derivative thereof, or Its fragments containing 15 or more consecutive bases, (I)
- Specimens used in the method of the present invention include specimens prepared from a subject's biological tissue (preferably, biliary tract tissue), body fluid such as blood, serum, plasma, urine, and the like. Specifically, an RNA-containing sample prepared from the tissue, a sample containing a polynucleotide further prepared therefrom, a body fluid such as blood, serum, plasma, urine, a part or all of a biological tissue of a subject, a biopsy, etc. Or a biological tissue extracted by surgery, etc., from which a specimen for measurement can be prepared.
- a subject refers to mammals such as, but not limited to, humans, monkeys, mice, rats, and the like, preferably humans.
- the steps can be changed according to the type of specimen used as a measurement target.
- RNA When RNA is used as a measurement object, detection of biliary tract cancer (cells) is performed, for example, by the following steps (a), (b) and (c): (A) binding RNA prepared from a specimen of a subject or a complementary polynucleotide (cDNA) transcribed therefrom to a polynucleotide in the kit or device of the present invention; (B) Measure RNA derived from a sample bound to the polynucleotide or cDNA synthesized from the RNA by hybridization using the polynucleotide as a nucleic acid probe or by quantitative RT-PCR using the polynucleotide as a primer. Step to do, (C) evaluating the presence or absence of biliary tract cancer (derived gene expression) based on the measurement result of (b) above, Can be included.
- steps (a), (b) and (c) (A) binding RNA prepared from a specimen of a subject or a complementary polynucleot
- hybridization methods for example, Northern blot method, Southern blot method, RT-PCR method, DNA chip analysis method, in situ hybridization method, Northern hybridization method, Southern hybridization method and the like can be used. .
- the presence or absence of each gene expression in RNA and the expression level thereof can be detected and measured by using the nucleic acid probe that can be used in the present invention.
- radioisotope nucleic acid probe complementary strand
- a signal derived from the formed DNA / RNA double-stranded label is detected with a radiation detector (BAS-1800II (Fuji Photo Film Co., Ltd.). ), Etc.) or a method of detecting and measuring with a fluorescence detector (STORM 865 (GE Healthcare) etc. can be exemplified).
- RNA and the expression level thereof can be detected and measured by using the above-described primers that can be used in the present invention.
- a pair of primers of the present invention (the cDNA described above) is prepared so that a region of each target gene can be amplified using a template prepared from RNA derived from biological tissue of a subject according to a conventional method.
- An example is a method of detecting a double-stranded DNA obtained by hybridizing with a cDNA with a normal strand and a reverse strand that bind to the DNA and performing a PCR method by a conventional method.
- the above PCR is performed using a primer previously labeled with a radioisotope or a fluorescent substance, the PCR product is electrophoresed on an agarose gel, and then is detected with ethidium bromide or the like.
- a method of detecting by staining the double-stranded DNA and a method of detecting the double-stranded DNA produced by transferring it to a nylon membrane or the like according to a conventional method and hybridizing with a labeled nucleic acid probe can be included.
- RNA chip or DNA chip in which the nucleic acid probe (single strand or double strand) of the present invention is attached to a substrate (solid phase) is used.
- the region where the nucleic acid probe is attached is called a probe spot, and the region where the nucleic acid probe is not attached is called a blank spot.
- a gene group immobilized on a substrate generally has a name such as a nucleic acid chip, a nucleic acid array, or a microarray, and a DNA or RNA array includes a DNA or RNA macroarray and a DNA or RNA microarray.
- the term “chip” includes all of them.
- 3D-Gene registered trademark
- Human miRNA Oligo chip Toray Industries, Inc.
- the measurement of the DNA chip is not limited.
- an image detector Teyphoon 9410 (GE Healthcare), 3D-Gene (registered trademark) scanner (Toray Industries, Inc.) or the like is used as a signal derived from the label of the nucleic acid probe.
- the method of detecting and measuring can be illustrated.
- stringent conditions refers to the extent to which a nucleic acid probe is larger than other sequences as described above (eg, average of background measurements + standard error of background measurements ⁇ 2 or more). In the measurement value) of the target sequence.
- Stringent conditions are determined by hybridization and subsequent washing conditions.
- the hybridization conditions are not limited, but for example, 30 to 60 ° C. and 1 to 24 hours in a solution containing SSC, surfactant, formamide, dextran sulfate, blocking agent and the like.
- 1 ⁇ SSC is an aqueous solution (pH 7.0) containing 150 mM sodium chloride and 15 mM sodium citrate, and the surfactant includes SDS (sodium dodecyl sulfate), Triton, or Tween.
- More preferable hybridization conditions include 3 to 10 ⁇ SSC and 0.1 to 1% SDS.
- Washing conditions after hybridization include, for example, a solution containing 0.5 ⁇ SSC at 30 ° C. and 0.1% SDS, and 0.2 at 30 ° C. There may be mentioned conditions such as continuous washing with a solution containing x SSC and 0.1% SDS and a 0.05 x SSC solution at 30 ° C. It is desirable that the complementary strand maintain a hybridized state with the target positive strand even when washed under such conditions.
- a complementary strand a strand consisting of a base sequence that is completely complementary to the target positive strand base sequence, and at least 80%, preferably at least 85%, more preferably, the strand. Examples include strands consisting of a base sequence having at least 90% or at least 95% identity, such as at least 98% or at least 99%.
- Examples of conditions for performing PCR using the polynucleotide fragment in the kit of the present invention as a primer include, for example, a PCR buffer having a composition such as 10 mM Tris-HCL (pH 8.3), 50 mM KCL, 1 to 2 mM MgCl 2.
- the Tm value calculated from the primer sequence +5 to 10 ° C. may be treated for 15 seconds to 1 minute.
- Calculation of gene expression level is not limited, but, for example, Statistical analysis of gene expression microarray data (Speed T., Chapman and HalliCensusChemistryChemistryChemistryChemistryChemistryChemistryChemistryChemistryChemistryChemistryChemistryChemistryChemistryChemistryCensus Et al., Blackwell publishing) can be used in the present invention.
- a spot can be regarded as a detection spot.
- the average value of the measured value of the blank spot can be regarded as the background, and can be subtracted from the measured value of the probe spot to obtain the gene expression level.
- the missing value of the gene expression level is excluded from the analysis target, preferably replaced with the minimum value of the gene expression level in each DNA chip, or more preferably 0.1 from the logarithmic value of the minimum value of the gene expression level. Can be replaced with the subtracted value.
- 20% or more preferably 50%, more preferably 80% or more of the number of measurement samples is 2 to the 6th power, preferably 2 to the 8th power, more preferably 2 to 10 Only genes having gene expression levels greater than or equal to the power can be selected for analysis. Examples of normalization of gene expression levels include, but are not limited to, global normalization and quantile normalization (Bolstad, B. M. et al., 2003, Bioinformatics, Vol. 19, p185-193).
- the present invention also measures the expression level of a target gene or gene in a specimen derived from a subject using the detection polynucleotide, kit, device (eg, chip), or a combination thereof of the present invention, and biliary tract cancer.
- a discriminant discriminant function
- the present invention further uses the detection polynucleotide, kit, device (for example, chip), or a combination thereof of the present invention to include a sample containing a gene derived from biliary tract cancer or a gene derived from biliary tract cancer.
- a first step of measuring in vitro the expression level of a target gene (target nucleic acid) in a plurality of specimens that are known to be determined or evaluated to be excluded, the target gene obtained in the first step The second step of creating a discriminant using the measured value of the expression level as a teacher sample, and the third step of measuring the expression level of the target gene in the specimen derived from the subject in vitro as in the first step Substituting the measured value of the expression level of the target gene obtained in the third step into the discriminant obtained in the second step, and based on the result obtained from the discriminant.
- determining or evaluating that the specimen contains a gene derived from biliary tract cancer or does not contain a gene derived from biliary tract cancer, wherein the target gene is the polynucleotide, kit or device A method is provided that is detectable by a detection polynucleotide contained in (eg, a chip).
- a discriminant can be created using Fisher's discriminant analysis, nonlinear discriminant analysis based on Mahalanobis distance, neural network, Support Vector Machine (SVM), etc., but is not limited to these.
- Linear discriminant analysis is a method for discriminating group membership using Equation 1 as a discriminant when the boundary of grouping is a straight line or a hyperplane.
- x is an explanatory variable
- w is a coefficient of the explanatory variable
- w 0 is a constant term.
- the value obtained by the discriminant is called a discriminant score, and the measured value of a newly given data set is substituted into the discriminant as an explanatory variable, and the grouping can be discriminated by the code of the discriminant score.
- Fisher's discriminant analysis which is a type of linear discriminant analysis, is a dimension reduction method for selecting a suitable dimension for class discrimination. Focusing on the variance of synthetic variables, the variance of data with the same label is minimized. To construct a highly discriminating synthetic variable (Venables, WN et al., Modern Applied Statistics with S. Fourth edition. Springer., 2002). In Fisher's discriminant analysis, a projection direction w that maximizes Equation 2 is obtained.
- ⁇ is the average of inputs
- ng is the number of data belonging to class g
- ⁇ g is the average of inputs of data belonging to class g.
- the numerator and denominator have inter-class variance and intra-class variance when the data is projected in the direction of the vector w, and the discriminant coefficient w i is obtained by maximizing this ratio.
- the Mahalanobis distance is calculated by Equation 3 in consideration of data correlation, and can be used as a nonlinear discriminant analysis for discriminating a group having a close Mahalanobis distance from each group as a belonging group.
- ⁇ is the center vector of each group
- S ⁇ 1 is the inverse matrix of the variance-covariance matrix of that group.
- the center vector is calculated from the explanatory variable x, and an average vector or a median vector can be used.
- a boundary surface called a hyperplane is used to correctly classify the data set into a known grouping, with specific data items in the data set with a known grouping as explanatory variables and the grouping to be classified as an objective variable. And determine a discriminant for classifying data using the boundary surface.
- the discriminant can determine the grouping by substituting the measured value of the newly given data set into the discriminant as an explanatory variable. Further, the discrimination result at this time may be a group to be classified, may be a probability of being classified into a group to be classified, or may be a distance from a hyperplane.
- a method for dealing with a non-linear problem a method is known in which a feature vector is non-linearly transformed into a higher dimension and linear identification is performed in the space.
- An expression in which the inner product of two elements in a non-linearly mapped space is expressed only by the input in the original space is called a kernel.
- a kernel a linear kernel, RBF (Radial Basis Function) Kernel and Gaussian kernel. It is possible to construct an optimum discriminant, that is, a discriminant by only calculating the kernel while actually calculating the feature in the mapped space while mapping in a high dimension by the kernel.
- C-support vector classification (C-SVC), a kind of SVM method, creates a hyperplane by learning with two explanatory variables to determine which group an unknown data set falls into (C. Cortes et al., 1995, Machine Learning, 20, p273-297).
- biliary tract cancer patients are divided into two groups: biliary tract cancer patients and healthy subjects.
- biliary histology can be used.
- a data set (hereinafter referred to as “learning sample group”) composed of comprehensive gene expression levels of the two groups of serum-derived specimens is prepared, and there is a clear difference in gene expression levels between the two groups.
- the discriminant by C-SVC is determined with the gene as the explanatory variable and the grouping as the target variable (eg, -1 and +1).
- Equation 4 is an objective function to be optimized, where e is all input vectors, y is an objective variable, a is a Lagrange undetermined multiplier vector, Q is a positive definite matrix, and C is a parameter for adjusting the constraint condition.
- Equation 5 is the discriminant finally obtained, and the group to which it belongs can be determined by the sign of the value obtained by the discriminant.
- x is a support vector
- y is a label indicating group membership
- a is a corresponding coefficient
- b is a constant term
- K is a kernel function.
- Equation 6 the RBF kernel defined by Equation 6 can be used.
- x represents a support vector
- ⁇ represents a kernel parameter that adjusts the complexity of the hyperplane.
- the subject-derived specimen contains and / or does not contain the expression of a target gene derived from biliary tract cancer, or the expression level thereof is compared with a control derived from a healthy subject.
- a neural network a k-neighbor method, a decision tree, a logistic regression analysis, or the like can be selected.
- the method of the present invention comprises, for example, the following steps (a), (b) and (c):
- B creating a discriminant of the above formulas 1 to 3, 5 and 6 from the measured value of the expression level measured in (a)
- (C) The expression level of the target gene in the specimen derived from the subject is measured using the detection polynucleotide, kit or device (for example, DNA chip) according to the present invention, and measured according to the discriminant created in (b).
- x in the formulas 1 to 3, 5 and 6 is an explanatory variable, and a value obtained by measuring a polynucleotide selected from the polynucleotides described in Section 2 above or a fragment thereof, and the like.
- the explanatory variable for discriminating between a biliary tract cancer patient and a healthy body of the present invention is, for example, a gene expression level selected from the following (1) to (2).
- a biliary tract cancer patient or a healthy subject measured by any one of DNAs containing 15 or more consecutive bases in the base sequence represented by any of SEQ ID NOs: 1-125, 466-478 or its complementary sequence Gene expression level in body serum.
- a biliary tract cancer patient or a healthy subject measured by any one of DNAs containing 15 or more consecutive bases in the base sequence represented by any of SEQ ID NOs: 1-125, 466-478 or its complementary sequence Gene expression level in body serum.
- the discriminant is created from a learning specimen group.
- the gene used as the explanatory variable of the discriminant is determined as follows. First, the comprehensive gene expression level of the biliary tract cancer patient group, which is the learning sample group, and the exhaustive gene expression level of the healthy body group are used as a data set. Using the P value of the Whitney U test or the P value of the Wilcoxon test, the magnitude of the difference in the expression level of each gene between the two groups is determined.
- Bonferroni correction for example, by multiplying the P value obtained by the test by the number of test repetitions, that is, the number of genes used in the analysis, and comparing it with the desired significance level, the first type of error in the entire test is obtained. Probability of occurrence can be suppressed.
- the absolute value of the median expression ratio (Fold change) of each gene expression level is calculated between the gene expression level of the biliary tract cancer patient group and the gene expression level of the healthy body group instead of the test, and the discriminant You may select the gene used for the explanatory variable.
- the ROC curve may be created using the gene expression levels of the biliary tract cancer patient group and the healthy body group, and the gene used for the explanatory variable of the discriminant may be selected based on the AUROC value.
- a discriminant that can be calculated by the above-described various methods is created using an arbitrary number of genes having a large difference in gene expression level obtained here.
- a method of constructing a discriminant that obtains the maximum discriminating accuracy for example, a method of constructing a discriminant with any combination of genes satisfying the significance level of the P value, or a gene used to create a discriminant, gene expression There is a method in which evaluation is repeated while increasing one by one in descending order of quantity difference (Furey TS. Et al., 2000, Bioinformatics, Vol. 16, p906-14).
- the gene expression level of another independent biliary tract cancer patient or healthy body is substituted into the explanatory variable, and the discrimination result of the group belonging to this independent biliary tract cancer patient or healthy body is calculated. That is, the diagnostic gene set that can detect more universal biliary tract cancer by evaluating the discriminant constructed using the found diagnostic gene set and the diagnostic gene set in an independent sample group, and Find ways to identify biliary tract cancer.
- the Split-sample method for evaluating the discriminating performance (generalization) of the discriminant is preferable to use the Split-sample method for evaluating the discriminating performance (generalization) of the discriminant. That is, the data set is divided into a learning sample group and a test sample group, the gene selection and the discriminant formula are made by statistical test in the learning sample group, and the test sample group is discriminated by the discriminant formula and the test sample group The accuracy, sensitivity, and specificity are calculated using the true group to which the belongs, and the discrimination performance is evaluated. On the other hand, without dividing the data set, the gene selection and discriminant formula are created by statistical test using all the samples, and the newly prepared sample is discriminated by the discriminant to improve accuracy, sensitivity, and specificity. It is also possible to calculate and evaluate the discrimination performance.
- the present invention relates to a polynucleotide for detection or disease diagnosis useful for diagnosis and treatment of biliary tract cancer, a method for detecting biliary tract cancer using the polynucleotide, and a biliary tract cancer detection kit and device containing the polynucleotide. I will provide a.
- the gene expressed in serum from patients whose biliary tract cancer was finally revealed by close examination such as computed tomography using a contrast medium, and biliary tract By comparing expressed genes in serum from patients who do not have cancer, a diagnostic gene set and discriminant that show an accuracy exceeding CEA and CA19-9 can be constructed.
- An arbitrary combination from one or more of the above-mentioned polynucleotides based on the base sequence represented by any of 126 to 148 or a complementary sequence thereof is taken as a diagnostic gene set.
- a discriminant is constructed by using the expression level of the diagnostic gene set in a specimen derived from a patient with a class I biliary tract cancer and a specimen derived from a class II healthy subject.
- RNA extraction from liquid sample kit was obtained from 300 ⁇ L of serum obtained from a total of 250 healthy subjects and 100 biliary tract cancer patients, including the above-mentioned learning sample group and test sample group. Using the RNA extraction reagent in (Toray Industries, Inc.), total RNA was obtained according to the protocol defined by the company.
- 3D-Gene (registered trademark) miRNA Labeling kit was obtained from total RNA obtained from the serum of a total of 250 healthy subjects and 100 biliary tract cancer patients. MiRNA was fluorescently labeled based on the protocol (ver 2.20) defined by the same company.
- 3D-Gene (registered trademark) Human miRNA Oligo chip equipped with a probe having a sequence complementary to 2,555 miRNAs among miRNAs registered in miRBBase release 20 as an oligo DNA chip ), Hybridization of miRNA in total RNA with the probe on the DNA chip and washing after hybridization were performed under stringent conditions based on the protocol established by the company.
- the DNA chip was scanned using a 3D-Gene (registered trademark) scanner (Toray Industries, Inc.), an image was acquired, and the fluorescence intensity was digitized with 3D-Gene (registered trademark) Extraction (Toray Industries, Inc.).
- the digitized fluorescence intensity is converted into a logarithmic value with a base of 2 to obtain the gene expression level, and the blank value is subtracted.
- the missing value is 0.1 from the logarithmic value of the minimum value of the gene expression level in each DNA chip. Replaced with the value obtained by subtracting.
- comprehensive miRNA gene expression levels were obtained for the serum of 100 biliary tract cancer patients and the serum of 150 healthy subjects.
- Example 1 ⁇ Selection of genetic markers using samples from the learning sample group and evaluation method for biliary tract cancer discrimination performance of single genetic markers using samples from the test sample group>
- a genetic marker for distinguishing biliary tract cancer from a healthy sample is selected from the learning sample group, and a single biliary tract is present for each selected genetic marker in the test sample group sample independent of the learning sample group. A method to evaluate the discrimination performance was examined.
- the miRNA expression levels of the learning sample group and the test sample group obtained in Reference Example 1 above were combined and normalized by quantile normalization.
- a diagnostic gene was selected using a group of learning samples.
- 2 6 6 or more genes in 50% or more of the samples Only genes with expression levels were selected.
- Bonferroni correction was performed on the P value obtained by the two-sided t-test assuming equal variance for each gene expression level, Genes satisfying p ⁇ 0.01 were obtained as genetic markers used as explanatory variables in the discriminant equation, and are listed in Table 2.
- a discriminant for discriminating the presence or absence of biliary tract cancer was created by Fisher's discriminant analysis. That is, among 125 genes selected in the learning sample group, a polynucleotide comprising a nucleotide sequence represented by any one of SEQ ID NOs: 1-125 newly found is input into Equation 2 to discriminate Table 3 shows the calculated accuracy, sensitivity, and specificity. Table 4 shows the discriminant coefficients and constant terms at that time.
- the accuracy, sensitivity, and specificity in the test sample group were calculated using the discriminant created above, and the discriminating performance of the selected polynucleotide was verified with an independent sample (Table 3).
- Table 3 For example, when the measured expression level of the nucleotide sequence represented by SEQ ID NO: 1 is compared between healthy subjects (100 people) and biliary tract cancer patients (67 people) in the learning sample group, the biliary tract cancer patient group is compared with the healthy body group. It was shown that the measured gene expression level was significantly lower (see the left side of FIG. 2), and this result was also reproducible in healthy subjects (50) and biliary tract cancer patients (33) in the test sample group (FIG. 2). 2 right).
- the measured value of gene expression in the biliary tract cancer patient group is significantly lower ( ⁇ ) or higher (+) than the healthy group (Table 2). These results were verified in the test sample group.
- the target of biliary tract cancer detection was calculated using a threshold value (5.69) for discriminating both groups set in the learning sample group, and as a result, true positive 33
- the detection performance of all the polynucleotides shown in SEQ ID NOs: 1-125 was calculated and listed in Table 3.
- SEQ ID NO: 1 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 34, 35, 36, 39, 40, 41, 42, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 60, 62 polynucleotides consisting of the base sequences represented by 62, 64, 65, 67, 68, 70, 74, 75, 76, 83, 84, 105, 107 are independently existing tumor markers CA19- Proof of distinguishing biliary tract cancer with sensitivity exceeding 9 It could be.
- polynucleotides having the base sequences represented by SEQ ID NOs: 1, 2, 3, 4, 10, 11, 12, 23, 64 are included in stages 0 and 1 ( All six biliary tract cancer specimens (including IA and IB) were correctly identified as biliary tract cancer. Therefore, these polynucleotides can also detect early biliary tract cancer and contribute to early diagnosis of biliary tract cancer.
- these polynucleotides were able to correctly discriminate any of the tumors that occupied the extrahepatic bile duct, intrahepatic bile duct, gallbladder, and papilla of the biliary tract in the test sample group.
- cancer of the lower bile duct, the nipple, and the intrahepatic bile duct, which are easily symptomatic with no symptoms, could be detected.
- Example 2 ⁇ Evaluation method of biliary tract cancer discrimination performance by combination of multiple genetic markers using test sample group samples>
- a method for evaluating biliary tract cancer discrimination performance by combining the genetic markers selected in Example 1 was examined. Specifically, Fisher's discriminant analysis is performed on 7,750 combinations of any two of the measured expression levels of the polynucleotide comprising the nucleotide sequence represented by SEQ ID NOs: 1-125 selected in Example 1. And constructed a discriminant to determine the presence or absence of biliary tract cancer.
- the discriminant created above the accuracy, sensitivity, and specificity in the test sample group were calculated, and the discriminating performance of the selected polynucleotide was verified with an independent sample.
- the biliary tract cancer discrimination of the test sample group was performed using a combination of the above 7,750 expression level measurement values of the polynucleotide, for example, a polynucleotide comprising the nucleotide sequences represented by SEQ ID NO: 2 and SEQ ID NO: 4
- the healthy specimen (50) in the test sample and the biliary tract cancer patient (33) are compared, and in the learning specimen group, the expression level of the healthy body group and the biliary tract cancer patient group is measured.
- a scatter plot in which the values were significantly separated was obtained (see FIG. 3 left), and this result was also reproducible in the test sample group (see FIG. 3 right).
- the healthy body group and the biliary tract cancer patient group Scatter plots were obtained that significantly separated the measured expression levels of these, and these results were verified in the test sample group.
- any two combinations of the measured expression levels of the polynucleotides consisting of the nucleotide sequences represented by SEQ ID NOs: 1-125 are capable of discriminating biliary tract cancers with a sensitivity exceeding CA19-9. Was proved.
- the measured expression level of the polynucleotide comprising the nucleotide sequence represented by SEQ ID NOs: 1-125 is 3, 4, 5, 6, 7, 8, 9, 10, or Even when more than that number is combined, a marker for detecting biliary tract cancer can be obtained with excellent sensitivity.
- the polynucleotides consisting of the nucleotide sequences represented by SEQ ID NOs: 1-125 selected in Example 1 were ranked in descending order of P values indicating statistical significance, and one was added from the top miRNA. Detection performance was calculated using a combination of one or more miRNAs.
- the sensitivity in the test sample group is 100% for one miRNA, 100% for 2 miRNAs, 100% for 3 miRNAs, 100% for 5 miRNAs, 100% for 10 miRNAs, It was 100% with 20 miRNAs, 100% with 50 miRNAs, and 100% with 100 miRNAs. Since these sensitivities are higher than those of existing blood tumor markers, it was shown that even when a plurality of miRNAs are combined, they can be excellent markers for detecting biliary tract cancer.
- the combination of a plurality of miRNAs is not limited to the case of adding in the order of statistical significance as described above, and any combination of a plurality of miRNAs can be used for detection of biliary tract cancer.
- CEA was defined as “ ⁇ ” when 5 ng / ml or less was “ ⁇ ”
- CA19-9 was defined as “ ⁇ ” when 37 U / ml or less, and “+” when exceeding those values.
- Example 3 Selection of genetic markers when all samples are used and evaluation method of biliary tract cancer discrimination performance of the captured genetic markers>
- the samples of the learning sample group and the test sample group used in Example 1 and Example 2 were integrated, and all the samples were used to select gene markers and evaluate their biliary tract cancer discrimination performance.
- the miRNA expression levels for the serum of 100 biliary tract cancer patients obtained in Reference Example 1 and the serum of 150 healthy subjects were normalized by quantile normalization.
- genes having a gene expression level of 2 6 or more in 50% or more of specimens in either the biliary tract cancer patient group or the healthy body group should be selected. Selected.
- Bonferroni correction was performed on the P value obtained by the two-sided t-test assuming equal variance for each gene expression level, A gene satisfying p ⁇ 0.01 was selected as a gene marker to be used as an explanatory variable of the discriminant and listed in Table 7.
- the measured expression level of the biliary tract cancer patient group is significantly lower than the healthy body group ( ⁇ ) or High (+) results (Table 7) were obtained and these results could be verified in the test sample group.
- ⁇ healthy body group
- Table 7 High (+) results
- Example 4 ⁇ Evaluation method of biliary tract cancer specific discrimination performance by combination of multiple genetic markers using test sample group samples>
- targeting the learning sample group of the sample group described in Reference Example 2 biliary tract cancer patients and healthy persons, colon cancer patients, stomach cancer patients, in the same manner as the method described in Example 1,
- the gene expression level of miRNA in serum was compared with the control group consisting of patients with esophageal cancer, liver cancer, and pancreaticobiliary benign disease, and additional diagnostic genetic markers were selected.
- one or more markers selected from the group consisting of the combination of the additional genetic marker for diagnosis (SEQ ID NOs: 466 to 478; see Table 1) and the genetic marker selected in Example 1 were used.
- the biliary tract cancer-specific discrimination performance was evaluated.
- the miRNA expression levels of the learning sample group and the test sample group obtained in Reference Example 2 above were combined and normalized by quantile normalization.
- Fisher's discriminant analysis is performed on 1 to 4 combinations including at least one expression level measurement value of any one of the polynucleotides consisting of the nucleotide sequences represented by SEQ ID NOs: 1 to 148 and 466 to 478, A discriminant was constructed to determine the presence or absence of biliary tract cancer.
- biliary tract cancer patient group as positive sample group, healthy body group, colon cancer patient group, stomach cancer patient group, esophageal cancer patient group, liver cancer patient group and pancreaticobiliary benign disease patient group as negative sample group
- the accuracy, sensitivity, and specificity in the test sample group were calculated using the discriminant created above, and the discriminating performance of the selected polynucleotide was verified with an independent sample.
- the biliary tract cancer could be specifically identified from other cancers.
- the polynucleotide that can specifically bind to the target marker include, for example, SEQ ID NOs: 1, 4, 5, 11, 12, 15, 23, 29, 39, 40, 54, 76, 79, 91, 103, 115.
- cancer type-specific polynucleotide group 1 A group consisting of a polynucleotide consisting of a base sequence represented by SEQ ID NOs: 4, 5, 12, 15 and 40 or a complementary sequence thereof among a plurality of polynucleotide combinations (cancer type-specific polynucleotide group 2)
- a combination comprising at least one polynucleotide selected from can specifically differentiate biliary tract cancer from other cancers with high accuracy Was capacity.
- the number of combinations of the above-mentioned cancer species-specific polynucleotides is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more.
- the discrimination accuracy of 80% or more could be shown in the combination of 4 or more.
- the discrimination accuracy when measured using a polynucleotide consisting of the base sequence represented by SEQ ID NO: 4 or its complementary sequence is shown below.
- the accuracy is 81.9% in the learning sample group and the accuracy is 76.9% in the test sample group. (Table 8).
- the maximum accuracy is 86.
- the discrimination accuracy when measured using a polynucleotide consisting of the base sequence represented by SEQ ID NO: 5 or its complementary sequence is shown below.
- the accuracy is 79.0% in the learning sample group, and the accuracy is 80.8% in the test sample group.
- Table 8 the maximum accuracy of 81.
- the accuracy was 86.5% in the test sample group of 9% (Table 9).
- the discrimination accuracy when measured using a polynucleotide consisting of the base sequence represented by SEQ ID NO: 12 or its complementary sequence is shown below.
- the accuracy is 80.6% in the learning sample group and the accuracy is 76.9% in the test sample group.
- Table 8 the maximum accuracy of 86.
- the accuracy was 35.9% in the test sample group (Table 9).
- the maximum accuracy in the learning sample group is 90.
- the accuracy was 91.7% in the test sample group of 2% (Table 10).
- the maximum accuracy when measurement is performed using a combination of four polynucleotides including at least one polynucleotide comprising the base sequence represented by SEQ ID NO: 12 or a complementary sequence thereof, the maximum accuracy of 93.
- the accuracy was 94.2% in the test sample group of 0% (Table 11).
- the discrimination accuracy when measured using a polynucleotide consisting of the base sequence represented by SEQ ID NO: 15 or its complementary sequence is shown below.
- the accuracy is 83.8% in the learning sample group, and the accuracy is 84.0% in the test sample group. (Table 8).
- the maximum accuracy of 89 was 89.1% in the test sample group with 5% (Table 9).
- the maximum accuracy in the learning sample group is 90.
- the accuracy was 92.3% in the test sample group with 5% (Table 10).
- the maximum accuracy when measurement is performed using a combination of four polynucleotides including at least one polynucleotide consisting of the base sequence represented by SEQ ID NO: 15 or a complementary sequence thereof, the maximum accuracy of 93.
- the accuracy was 94.2% in the test sample group of 0% (Table 11).
- the discrimination accuracy when measured using a polynucleotide consisting of the base sequence represented by SEQ ID NO: 40 or its complementary sequence is shown below.
- the accuracy was 80.0% in the learning sample group, and the accuracy was 76.9% in the test sample group. (Table 8).
- the maximum accuracy of 81 when measured using a combination of two polynucleotides including at least one polynucleotide consisting of the base sequence represented by SEQ ID NO: 40 or a complementary sequence thereof, the maximum accuracy of 81. The accuracy was 86.5% in the test sample group of 9% (Table 9).
- the maximum accuracy of 86 when measurement is performed using a combination of three polynucleotides including at least one polynucleotide comprising the nucleotide sequence represented by SEQ ID NO: 40 or a complementary sequence thereof, the maximum accuracy of 86. The accuracy was 89.7% in 7% and the test sample group (Table 10). For example, when measurement is performed using a combination of four polynucleotides including at least one polynucleotide consisting of the base sequence represented by SEQ ID NO: 40 or a complementary sequence thereof, the maximum accuracy of 91. The accuracy was 91.7% in the test sample group of 4% (Table 11).
- the sensitivity of CEA is 31.3%
- the sensitivity of CA19-9 is 68.2%
- the sensitivity of CEA is 33.3%
- the sensitivity of CA19-9 is only 59.4%. None of the markers were found to be useful for the detection of biliary tract cancer (Table 5).
- biliary tract cancer can be detected with higher sensitivity than existing tumor markers, so that early removal of cancerous part by surgery is performed. As a result, it becomes possible to improve the 5-year survival rate and reduce the recurrence rate.
- biliary tract cancer can be effectively detected by a simple and inexpensive method, early detection, diagnosis and treatment of biliary tract cancer are possible.
- biliary tract cancer can be detected in a minimally invasive manner using patient blood, so that biliary tract cancer can be detected easily and quickly.
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Abstract
Description
すなわち、本発明は、以下の特徴を有する。
(1)胆道がんマーカーである、miR-125a-3p、miR-6893-5p、miR-204-3p、miR-4476、miR-4294、miR-150-3p、miR-6729-5p、miR-7641、miR-6765-3p、miR-6820-5p、miR-575、miR-6836-3p、miR-1469、miR-663a、miR-6075、miR-4634、miR-423-5p、miR-4454、miR-7109-5p、miR-6789-5p、miR-6877-5p、miR-4792、miR-4530、miR-7975、miR-6724-5p、miR-8073、miR-7977、miR-1231、miR-6799-5p、miR-615-5p、miR-4450、miR-6726-5p、miR-6875-5p、miR-4734、miR-16-5p、miR-602、miR-4651、miR-8069、miR-1238-5p、miR-6880-5p、miR-8072、miR-4723-5p、miR-4732-5p、miR-6125、miR-6090、miR-7114-5p、miR-564、miR-451a、miR-3135b、miR-4497、miR-4665-5p、miR-3622a-5p、miR-6850-5p、miR-6821-5p、miR-5100、miR-6872-3p、miR-4433-3p、miR-1227-5p、miR-3188、miR-7704、miR-3185、miR-1908-3p、miR-6781-5p、miR-6805-5p、miR-8089、miR-665、miR-4486、miR-6722-3p、miR-1260a、miR-4707-5p、miR-6741-5p、miR-1260b、miR-1246、miR-6845-5p、miR-4638-5p、miR-6085、miR-1228-3p、miR-4534、miR-5585-3p、miR-4741、miR-4433b-3p、miR-197-5p、miR-718、miR-4513、miR-4446-3p、miR-619-5p、miR-6816-5p、miR-6778-5p、miR-24-3p、miR-1915-3p、miR-4665-3p、miR-4449、miR-6889-5p、miR-486-3p、miR-7113-3p、miR-642a-3p、miR-7847-3p、miR-6768-5p、miR-1290、miR-7108-5p、miR-92b-5p、miR-663b、miR-3940-5p、miR-4467、miR-6858-5p、miR-4417、miR-3665、miR-4736、miR-4687-3p、miR-1908-5p、miR-5195-3p、miR-4286、miR-3679-3p、miR-6791-5p、miR-1202、miR-3656、miR-4746-3p、miR-3184-5p、miR-3937、miR-6515-3p、miR-6132、miR-187-5p、miR-7111-5p、miR-5787、miR-6779-5p、miR-4516、miR-4649-5p、miR-760、miR-3162-5p、miR-3178、miR-940、miR-4271、miR-6769b-5p、miR-4508、miR-6826-5p、miR-6757-5p、miR-3131、及び、miR-1343-3pからなる群から選択される少なくとも1つ以上のポリヌクレオチドと特異的に結合可能な核酸を含む、胆道がんの検出用キット。
(a)配列番号1~125、466~478のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(b)配列番号1~125、466~478のいずれかで表される塩基配列を含むポリヌクレオチド、
(c)配列番号1~125、466~478のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(d)配列番号1~125、466~478のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列に相補的な塩基配列を含むポリヌクレオチド、及び
(e)前記(a)~(d)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド、
からなる群から選択されるポリヌクレオチドである、(1)又は(2)に記載のキット。
(f)配列番号126~148のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(g)配列番号126~148のいずれかで表される塩基配列を含むポリヌクレオチド、
(h)配列番号126~148のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(i)配列番号126~148のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列に相補的な塩基配列を含むポリヌクレオチド、及び
(j)前記(f)~(i)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド、
からなる群から選択されるポリヌクレオチドである、(4)又は(5)に記載のキット。
(a)配列番号1~125、466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(b)配列番号1~125、466~478のいずれかで表される塩基配列を含むポリヌクレオチド、
(c)配列番号1~125、466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(d)配列番号1~125、466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列を含むポリヌクレオチド、及び
(e)前記(a)~(d)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド、
からなる群から選択されるポリヌクレオチドである、(8)又は(9)に記載のデバイス。
(f)配列番号126~148のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(g)配列番号126~148のいずれかで表される塩基配列を含むポリヌクレオチド、
(h)配列番号126~148のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(i)配列番号126~148のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列を含むポリヌクレオチド、及び
(j)前記(f)~(i)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド、
からなる群から選択されるポリヌクレオチドである、(11)又は(12)に記載のデバイス。
(15)前記ハイブリダイゼーション技術が、核酸アレイ技術である、(14)に記載のデバイス。
(16)前記デバイスが、(8)又は(9)に記載のすべての胆道がんマーカーから選択される少なくとも2つ以上のポリヌクレオチドのそれぞれと特異的に結合可能な少なくとも2つ以上の核酸を含む、(8)~(15)のいずれかに記載のデバイス。
(18)前記被験体が、ヒトである、(17)に記載の方法。
(19)前記検体が、血液、血清又は血漿である、(17)又は(18)に記載の方法。
本明細書中で使用する用語は、以下の定義を有する。
1.胆道がんの標的核酸
本発明の上記定義の胆道がん検出用の核酸プローブ又はプライマーを使用して、胆道がん又は胆道がん細胞の存在及び/又は不存在を検出するための、胆道がんマーカーとしての主要な標的核酸には、hsa-miR-125a-3p、hsa-miR-6893-5p、hsa-miR-204-3p、hsa-miR-4476、hsa-miR-4294、hsa-miR-150-3p、hsa-miR-6729-5p、hsa-miR-7641、hsa-miR-6765-3p、hsa-miR-6820-5p、hsa-miR-575、hsa-miR-6836-3p、hsa-miR-1469、hsa-miR-663a、hsa-miR-6075、hsa-miR-4634、hsa-miR-423-5p、hsa-miR-4454、hsa-miR-7109-5p、hsa-miR-6789-5p、hsa-miR-6877-5p、hsa-miR-4792、hsa-miR-4530、hsa-miR-7975、hsa-miR-6724-5p、hsa-miR-8073、hsa-miR-7977、hsa-miR-1231、hsa-miR-6799-5p、hsa-miR-615-5p、hsa-miR-4450、hsa-miR-6726-5p、hsa-miR-6875-5p、hsa-miR-4734、hsa-miR-16-5p、hsa-miR-602、hsa-miR-4651、hsa-miR-8069、hsa-miR-1238-5p、hsa-miR-6880-5p、hsa-miR-8072、hsa-miR-4723-5p、hsa-miR-4732-5p、hsa-miR-6125、hsa-miR-6090、hsa-miR-7114-5p、hsa-miR-564、hsa-miR-451a、hsa-miR-3135b、hsa-miR-4497、hsa-miR-4665-5p、hsa-miR-3622a-5p、hsa-miR-6850-5p、hsa-miR-6821-5p、hsa-miR-5100、hsa-miR-6872-3p、hsa-miR-4433-3p、hsa-miR-1227-5p、hsa-miR-3188、hsa-miR-7704、hsa-miR-3185、hsa-miR-1908-3p、hsa-miR-6781-5p、hsa-miR-6805-5p、hsa-miR-8089、hsa-miR-665、hsa-miR-4486、hsa-miR-6722-3p、hsa-miR-1260a、hsa-miR-4707-5p、hsa-miR-6741-5p、hsa-miR-1260b、hsa-miR-1246、hsa-miR-6845-5p、hsa-miR-4638-5p、hsa-miR-6085、hsa-miR-1228-3p、hsa-miR-4534、hsa-miR-5585-3p、hsa-miR-4741、hsa-miR-4433b-3p、hsa-miR-197-5p、hsa-miR-718、hsa-miR-4513、hsa-miR-4446-3p、hsa-miR-619-5p、hsa-miR-6816-5p、hsa-miR-6778-5p、hsa-miR-24-3p、hsa-miR-1915-3p、hsa-miR-4665-3p、hsa-miR-4449、hsa-miR-6889-5p、hsa-miR-486-3p、hsa-miR-7113-3p、hsa-miR-642a-3p、hsa-miR-7847-3p、hsa-miR-6768-5p、hsa-miR-1290、hsa-miR-7108-5p、hsa-miR-92b-5p、hsa-miR-663b、hsa-miR-3940-5p、hsa-miR-4467、hsa-miR-6858-5p、hsa-miR-4417、hsa-miR-3665、hsa-miR-4736、hsa-miR-4687-3p、hsa-miR-1908-5p、hsa-miR-5195-3p、hsa-miR-4286、hsa-miR-3679-3p、hsa-miR-6791-5p、hsa-miR-1202、hsa-miR-3656、hsa-miR-4746-3p、hsa-miR-3184-5p、hsa-miR-3937、hsa-miR-6515-3p、hsa-miR-6132、hsa-miR-187-5p、hsa-miR-7111-5p、hsa-miR-5787、hsa-miR-6779-5p、hsa-miR-4516、hsa-miR-4649-5p、hsa-miR-760、hsa-miR-3162-5p、hsa-miR-3178、hsa-miR-940、hsa-miR-4271、hsa-miR-6769b-5p、hsa-miR-4508、hsa-miR-6826-5p、hsa-miR-6757-5p、hsa-miR-3131、及び、hsa-miR-1343-3pからなる群から選択される少なくとも1つ以上のmiRNAを用いることができる。さらにこれらのmiRNAと組み合わせることができる他の胆道がんマーカー、すなわち、hsa-miR-6808-5p、hsa-miR-6774-5p、hsa-miR-4656、hsa-miR-6806-5p、hsa-miR-1233-5p、hsa-miR-328-5p、hsa-miR-4674、hsa-miR-2110、hsa-miR-6076、hsa-miR-3619-3p、hsa-miR-92a-2-5p、hsa-miR-128-1-5p、hsa-miR-638、hsa-miR-2861、hsa-miR-371a-5p、hsa-miR-211-3p、hsa-miR-1273g-3p、hsa-miR-1203、hsa-miR-122-5p、hsa-miR-4258、hsa-miR-4484、hsa-miR-4648及びhsa-miR-6780b-5pからなる群から選択される少なくとも1つ以上のmiRNAも標的核酸として好ましく用いることができる。
本発明においては、上記の胆道がんマーカーとしての標的核酸に特異的に結合可能な核酸を、胆道がんを検出又は診断するための核酸、例えば核酸プローブ又はプライマーとして用いることができる。
(a)配列番号1~125、466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(b)配列番号1~125、466~478のいずれかで表される塩基配列を含むポリヌクレオチド、
(c)配列番号1~125、466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(d)配列番号1~125、466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列を含むポリヌクレオチド、並びに、
(e)前記(a)~(d)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド。
(f)配列番号126~148のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(g)配列番号126~148のいずれかで表される塩基配列を含むポリヌクレオチド、
(h)配列番号126~148のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(i)配列番号126~148のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列を含むポリヌクレオチド、並びに、
(j)前記(f)~(i)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド。
本発明はまた、胆道がんマーカーである標的核酸を測定するための、本発明において核酸プローブ又はプライマーとして使用可能なポリヌクレオチド(これには、変異体、断片、又は誘導体を含みうる。;以下、検出用ポリヌクレオチドと称することがある)の1つ又は複数を含む胆道がん検出用キット又はデバイスを提供する。
miR-125a-3p、miR-6893-5p、miR-204-3p、miR-4476、miR-4294、miR-150-3p、miR-6729-5p、miR-7641、miR-6765-3p、miR-6820-5p、miR-575、miR-6836-3p、miR-1469、miR-663a、miR-6075、miR-4634、miR-423-5p、miR-4454、miR-7109-5p、miR-6789-5p、miR-6877-5p、miR-4792、miR-4530、miR-7975、miR-6724-5p、miR-8073、miR-7977、miR-1231、miR-6799-5p、miR-615-5p、miR-4450、miR-6726-5p、miR-6875-5p、miR-4734、miR-16-5p、miR-602、miR-4651、miR-8069、miR-1238-5p、miR-6880-5p、miR-8072、miR-4723-5p、miR-4732-5p、miR-6125、miR-6090、miR-7114-5p、miR-564、miR-451a、miR-3135b、miR-4497、miR-4665-5p、miR-3622a-5p、miR-6850-5p、miR-6821-5p、miR-5100、miR-6872-3p、miR-4433-3p、miR-1227-5p、miR-3188、miR-7704、miR-3185、miR-1908-3p、miR-6781-5p、miR-6805-5p、miR-8089、miR-665、miR-4486、miR-6722-3p、miR-1260a、miR-4707-5p、miR-6741-5p、miR-1260b、miR-1246、miR-6845-5p、miR-4638-5p、miR-6085、miR-1228-3p、miR-4534、miR-5585-3p、miR-4741、miR-4433b-3p、miR-197-5p、miR-718、miR-4513、miR-4446-3p、miR-619-5p、miR-6816-5p、miR-6778-5p、miR-24-3p、miR-1915-3p、miR-4665-3p、miR-4449、miR-6889-5p、miR-486-3p、miR-7113-3p、miR-642a-3p、miR-7847-3p、miR-6768-5p、miR-1290、miR-7108-5p、miR-92b-5p、miR-663b、miR-3940-5p、miR-4467、miR-6858-5p、miR-4417、miR-3665、miR-4736、miR-4687-3p、miR-1908-5p、miR-5195-3p、miR-4286、miR-3679-3p、miR-6791-5p、miR-1202、miR-3656、miR-4746-3p、miR-3184-5p、miR-3937、miR-6515-3p、miR-6132、miR-187-5p、miR-7111-5p、miR-5787、miR-6779-5p、miR-4516、miR-4649-5p、miR-760、miR-3162-5p、miR-3178、miR-940、miR-4271、miR-6769b-5p、miR-4508、miR-6826-5p、miR-6757-5p、miR-3131、及び、miR-1343-3p。
(1)配列番号1~125、466~478のいずれかで表される塩基配列においてuがtである塩基配列又はその相補的配列において、15以上の連続した塩基を含むポリヌクレオチド。
(2)配列番号126~148のいずれかで表される塩基配列においてuがtである塩基配列又はその相補的配列において、15以上の連続した塩基を含むポリヌクレオチド。
(2)配列番号4、5、12、76(マーカー:miR-4476、miR-4294、miR-6836-3p、miR-6085)の組み合わせ
(3)配列番号4、5、12、115(マーカー:miR-4476、miR-4294、miR-6836-3p、miR-1202)の組み合わせ
(4)配列番号4、12、15、474(マーカー:miR-4476、miR-6836-3p、miR-6075、miR-4508)の組み合わせ
(5)配列番号4、15、29、115(マーカー:miR-4476、miR-6075、miR-6799-5p、miR-1202)の組み合わせ
(2)配列番号5、76、54、115(マーカー:hsa-miR-4294、hsa-miR-6085、hsa-miR-6821-5p、hsa-miR-1202)の組み合わせ
(3)配列番号5、23、12、115(マーカー:hsa-miR-4294、hsa-miR-4530、hsa-miR-6836-3p、hsa-miR-1202)の組み合わせ
(4)配列番号5、12、115、91(マーカー:hsa-miR-4294、hsa-miR-6836-3p、hsa-miR-1202、hsa-miR-4665-3p)の組み合わせ
(5)配列番号5、1、23、4(マーカー:hsa-miR-4294、hsa-miR-125a-3p、hsa-miR-4530、hsa-miR-4476)の組み合わせ
(2)配列番号12、15、23、115(マーカー:miR-6836-3p、miR-6075、miR-4530、miR-1202)の組み合わせ
(3)配列番号5、12、115、469(マーカー:miR-4294、miR-6836-3p、miR-3162-5p、miR-1202)の組み合わせ
(4)配列番号5、12、115、472(マーカー:miR-4294、miR-6836-3p、miR-1202、miR-4271)の組み合わせ
(5)配列番号5、12、76、115(マーカー:miR-4294、miR-6085、miR-1202、miR-6836-3p)の組み合わせ
(2)配列番号15、12、11、143(マーカー:hsa-miR-6075、hsa-miR-6836-3p、hsa-miR-575、hsa-miR-1203)の組み合わせ
(3)配列番号15、76、121、39(マーカー:hsa-miR-6075、hsa-miR-6085、hsa-miR-6132、hsa-miR-1238-5p)の組み合わせ
(4)配列番号15、76、54、121(マーカー:hsa-miR-6075、hsa-miR-6085、hsa-miR-6821-5p、hsa-miR-6132)の組み合わせ
(5)配列番号15、40、1、23(マーカー:hsa-miR-6075、hsa-miR-6880-5p、hsa-miR-125a-3p、hsa-miR-4530)の組み合わせ
(2)配列番号12、23、40、466(マーカー:miR-6836-3p、miR-4530、miR-6880-5p、miR-4516)の組み合わせ
(3)配列番号12、23、40、134(マーカー:miR-6836-3p、miR-4530、miR-6880-5p、miR-6076)の組み合わせ
(4)配列番号15、40、121、134(マーカー:miR-6075、miR-6880-5p、miR-6132、miR-6076)の組み合わせ
(5)配列番号15、40、54、76(マーカー:miR-6075、miR-6880-5p、miR-6821-5p、miR-6085)の組み合わせ
本発明はさらに、上記3.で説明した本発明のキット又はデバイス(本発明で使用可能な上記の核酸を含む。)を用いて、検体中の、以下の群:miR-125a-3p、miR-6893-5p、miR-204-3p、miR-4476、miR-4294、miR-150-3p、miR-6729-5p、miR-7641、miR-6765-3p、miR-6820-5p、miR-575、miR-6836-3p、miR-1469、miR-663a、miR-6075、miR-4634、miR-423-5p、miR-4454、miR-7109-5p、miR-6789-5p、miR-6877-5p、miR-4792、miR-4530、miR-7975、miR-6724-5p、miR-8073、miR-7977、miR-1231、miR-6799-5p、miR-615-5p、miR-4450、miR-6726-5p、miR-6875-5p、miR-4734、miR-16-5p、miR-602、miR-4651、miR-8069、miR-1238-5p、miR-6880-5p、miR-8072、miR-4723-5p、miR-4732-5p、miR-6125、miR-6090、miR-7114-5p、miR-564、miR-451a、miR-3135b、miR-4497、miR-4665-5p、miR-3622a-5p、miR-6850-5p、miR-6821-5p、miR-5100、miR-6872-3p、miR-4433-3p、miR-1227-5p、miR-3188、miR-7704、miR-3185、miR-1908-3p、miR-6781-5p、miR-6805-5p、miR-8089、miR-665、miR-4486、miR-6722-3p、miR-1260a、miR-4707-5p、miR-6741-5p、miR-1260b、miR-1246、miR-6845-5p、miR-4638-5p、miR-6085、miR-1228-3p、miR-4534、miR-5585-3p、miR-4741、miR-4433b-3p、miR-197-5p、miR-718、miR-4513、miR-4446-3p、miR-619-5p、miR-6816-5p、miR-6778-5p、miR-24-3p、miR-1915-3p、miR-4665-3p、miR-4449、miR-6889-5p、miR-486-3p、miR-7113-3p、miR-642a-3p、miR-7847-3p、miR-6768-5p、miR-1290、miR-7108-5p、miR-92b-5p、miR-663b、miR-3940-5p、miR-4467、miR-6858-5p、miR-4417、miR-3665、miR-4736、miR-4687-3p、miR-1908-5p、miR-5195-3p、miR-4286、miR-3679-3p、miR-6791-5p、miR-1202、miR-3656、miR-4746-3p、miR-3184-5p、miR-3937、miR-6515-3p、miR-6132、miR-187-5p、miR-7111-5p、miR-5787及びmiR-6779-5pから選択される胆道がん由来の遺伝子の発現量、並びに場合により、以下の群:miR-6808-5p、miR-6774-5p、miR-4656、miR-6806-5p、miR-1233-5p、miR-328-5p、miR-4674、miR-2110、miR-6076、miR-3619-3p、miR-92a-2-5p、miR-128-1-5p、miR-638、miR-2861、miR-371a-5p、miR-211-3p、miR-1273g-3p、miR-1203、miR-122-5p、miR-4258、miR-4484、miR-4648、miR-6780b-5p、miR-4516、miR-4649-5p、miR-760、miR-3162-5p、miR-3178、miR-940、miR-4271、miR-6769b-5p、miR-4508、miR-6826-5p、miR-6757-5p、miR-3131、及び、miR-1343-3pから選択される胆道がん由来の遺伝子の発現量、の1つ以上で表される胆道がん由来の遺伝子の発現量、をin vitroで測定し、さらに、胆道がんの罹患が疑われる被験体と、健常体(非胆道がん患者を含む)とから採取した血液、血清、血漿等の検体について、検体中の上記遺伝子の発現量と、健常体の対照発現量とを用いて、例えば両発現量を比較して、当該検体中の標的核酸の発現量に統計学的に有意に差がある場合、被験体が、胆道がんに罹患していると評価することを含む、胆道がんの検出方法を提供する。
(a)被験体由来の検体を、in vitroで、本発明のキット又はデバイス中のポリヌクレオチドと接触させるステップ、
(b)検体中の標的核酸の発現量を、上記ポリヌクレオチドを核酸プローブ又はプライマーとして用いて測定するステップ、
(c)(b)の結果をもとに、当該被験体中の胆道がん(細胞)の存在又は不存在を評価するステップ、
を含むことができる。
(a)配列番号1~125、466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(b)配列番号1~125、466~478のいずれかで表される塩基配列を含むポリヌクレオチド、
(c)配列番号1~125、466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(d)配列番号1~125、466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列を含むポリヌクレオチド、及び
(e)前記(a)~(d)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド、
からなる群から選択される。
(f)配列番号126~148のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(g)配列番号126~148のいずれかで表される塩基配列を含むポリヌクレオチド、
(h)配列番号126~148のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(i)配列番号126~148のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列を含むポリヌクレオチド、及び
(j)前記(f)~(i)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド、
からなる群から選択される。
(a)被験体の検体から調製されたRNA又はそれから転写された相補的ポリヌクレオチド(cDNA)を、本発明のキット又はデバイス中のポリヌクレオチドと結合させるステップ、
(b)当該ポリヌクレオチドに結合した検体由来のRNA又は当該RNAから合成されたcDNAを、上記ポリヌクレオチドを核酸プローブとして用いるハイブリダイゼーションによって、あるいは、上記ポリヌクレオチドをプライマーとして用いる定量RT-PCRによって測定するステップ、
(c)上記(b)の測定結果に基づいて、胆道がん(由来の遺伝子の発現)の存在又は不存在を評価するステップ、
を含むことができる。
(a)胆道がん患者由来の胆道がん由来遺伝子を含む組織及び/又は健常体由来の胆道がん由来遺伝子を含まない組織であることが既に知られている検体中の標的遺伝子の発現量を、本発明による検出用ポリヌクレオチド、キット又はデバイス(例えばDNAチップ)を用いて測定するステップ、
(b)(a)で測定された発現量の測定値から、上記の式1~3、5及び6の判別式を作成するステップ、
(c)被験体由来の検体中の当該標的遺伝子の発現量を、本発明による検出用ポリヌクレオチド、キット又はデバイス(例えばDNAチップ)を用いて測定し、(b)で作成した判別式に測定値を代入して、得られた結果に基づいて検体が胆道がん由来の標的遺伝子を含むこと及び/又は含まないことを決定又は評価する、あるいはその発現量を健常体由来の対照と比較し評価する、ステップ、
を含むことができる。ここで、式1~3、5及び6の式中のxは説明変数であり、上記2節に記載したポリヌクレオチド類から選択されるポリヌクレオチド又はその断片等を測定することによって得られる値を含み、具体的には本発明の胆道がん患者と健常体を判別するための説明変数は、例えば下記の(1)~(2)より選択される遺伝子発現量である。
(1)配列番号1~125、466~478のいずれかで表される塩基配列又はその相補的配列において、15以上の連続した塩基を含むDNAのいずれかによって測定される胆道がん患者もしくは健常体の血清における遺伝子発現量。
(2)配列番号126~148のいずれかで表される塩基配列又はその相補的配列において、15以上の連続した塩基を含むDNAのいずれかによって測定される胆道がん患者もしくは健常体の血清における遺伝子発現量。
<胆道がん患者と健常体の検体の採取>
インフォームドコンセントを得た健常体100人と胆道以外に原発がんが認められていない胆道がん患者67人(ステージIAが1例、ステージIBが8例、ステージIIが8例、ステージIIAが3例、ステージIIBが5例、ステージIIIが14例、ステージIIIBが2例、ステージIVaが1例、ステージIVbが25例)からベノジェクトII真空採血管VP-AS109K60(テルモ株式会社)を用いてそれぞれ血清を採取し、学習検体群とした。同様に、インフォームドコンセントを得た健常体50人と胆道以外に原発がんが認められていない胆道がん患者33人(ステージ0が1例、ステージIが2例、ステージIAが1例、ステージIBが2例、ステージIIが2例、ステージIIAが5例、ステージIIBが4例、ステージIIIが5例、ステージIVが1例、ステージIVaが1例、ステージIVbが9例)からベノジェクトII真空採血管VP-AS109K60(テルモ株式会社)を用いてそれぞれ血清を採取し、テスト検体群とした。
検体として上記学習検体群、テスト検体群合わせて健常体150人と胆道がん患者100人の合計250人からそれぞれ得られた血清300μLから、3D-Gene(登録商標)RNA extraction reagent from liquid sample kit(東レ株式会社)中のRNA抽出用試薬を用いて、同社の定めるプロトコールに従ってtotal RNAを得た。
検体として上記学習検体群、テスト検体群合わせて健常体150人と胆道がん患者100人の合計250人の血清から得たtotal RNAに対して、3D-Gene(登録商標) miRNA Labeling kit(東レ株式会社)を用いて同社が定めるプロトコール(ver2.20)に基づいてmiRNAを蛍光標識した。オリゴDNAチップとして、miRBase release 20に登録されているmiRNAの中で、2,555種のmiRNAと相補的な配列を有するプローブを搭載した3D-Gene(登録商標) Human miRNA Oligo chip(東レ株式会社)を用い、同社が定めるプロトコールに基づいてストリンジェントな条件で、total RNA中のmiRNAとDNAチップ上のプローブとのハイブリダイゼーション及びハイブリダイゼーション後の洗浄を行った。DNAチップを3D-Gene(登録商標)スキャナー(東レ株式会社)を用いてスキャンし、画像を取得して3D-Gene(登録商標)Extraction(東レ株式会社)にて蛍光強度を数値化した。数値化された蛍光強度を、底が2の対数値に変換して遺伝子発現量とし、ブランク値の減算を行い、欠損値は各DNAチップにおける遺伝子発現量の最小値の対数値から0.1を減算した値で置換した。その結果、胆道がん患者100人の血清及び健常体150人の血清に対する、網羅的なmiRNAの遺伝子発現量を得た。数値化されたmiRNAの遺伝子発現量を用いた計算及び統計解析は、R言語3.0.2(R Development Core Team (2013). R: A language and environment for statistical computing. R Foundation for Statistical Computing, URL http://www.R-project.org/.)及びMASSパッケージ7.3-30(Venables, W. N. & Ripley, B. D. (2002) Modern Applied Statistics with S. Fourth Edition. Springer, New York. ISBN 0-387-95457-0)を用いて実施した。
<他のがんと良性疾患の検体の採取>
インフォームドコンセントを得た他の臓器にがんが認められていない大腸がん患者35人、胃がん患者37人、食道がん患者32人、肝がん患者38人及び膵胆道良性疾患患者13人からベノジェクトII真空採血管VP-AS109K60(テルモ株式会社)を用いてそれぞれ血清を採取し、参考例1の胆道がん患者67人(ステージ0が1例、ステージIが2例、ステージIAが1例、ステージIBが4例、ステージIIが8例、ステージIIAが4例、ステージIIBが6例、ステージIIIが14例、ステージIIIBが1例、ステージIVが25例、ステージIVaが1例)と健常体93人とを合わせて学習検体群とした。同様に、インフォームドコンセントを得た他の臓器にがんが認められていない大腸がん患者15人、胃がん患者13人、食道がん患者18人、肝がん患者12人及び膵胆道良性疾患患者8人からベノジェクトII真空採血管VP-AS109K60(テルモ株式会社)を用いてそれぞれ血清を採取し、参考例1の胆道がん患者33人(ステージIAが1例、ステージIBが6例、ステージIIが2例、ステージIIAが4例、ステージIIBが3例、ステージIIIが5例、ステージIIIBが1例、ステージIVが11例)、健常体57人と合わせてテスト検体群とした。以降のtotalRNAの抽出並びに遺伝子発現量の測定及び解析は参考例1と同様に行った。
<学習検体群の検体を用いた遺伝子マーカーの選定とテスト検体群の検体を用いた単独の遺伝子マーカーの胆道がん判別性能の評価方法>
本実施例では、学習検体群から胆道がんを健常体と判別するための遺伝子マーカーを選定し、学習検体群とは独立したテスト検体群の検体において選定した遺伝子マーカーについてそれぞれ単独での胆道がん判別性能を評価する方法を検討した。
<テスト検体群検体を用いた複数の遺伝子マーカーの組合せによる胆道がん判別性能の評価方法>
本実施例では、実施例1で選定された遺伝子マーカーを組合せて胆道がん判別性能を評価する方法を検討した。具体的には、実施例1において選択された配列番号1~125で表される塩基配列からなるポリヌクレオチドの発現量測定値のうちいずれか2個の組み合わせ7,750通りについてフィッシャーの判別分析を行い、胆道がんの存在の有無を判別する判別式を構築した。次に、上記で作成した判別式を用いてテスト検体群における精度・感度・特異度を算出し、選定されたポリヌクレオチドの判別性能を独立した検体で検証した。上記7,750通りのポリヌクレオチドの発現量測定値の組合せを用いてテスト検体群の胆道がん判別を実施したところ、例えば、配列番号2と配列番号4で表される塩基配列からなるポリヌクレオチドの発現量測定値を用いて、テスト検体中の健常体(50人)と胆道がん患者(33人)で比較した場合、学習検体群では健常体群と胆道がん患者群の発現量測定値が有意に分離する散布図が得られ(図3左参照)、更にこの結果はテスト検体群でも再現ができた(図3右参照)。同様に、新規に見出された配列番号1~125で表される塩基配列からなるポリヌクレオチドの発現量測定値のいずれか2個の他の組み合わせにおいても、健常体群と胆道がん患者群の発現量測定値を有意に分離する散布図が得られ、これらの結果はテスト検体群で検証ができた。また、例えばこの配列番号2と配列番号4で示される塩基配列に関し、学習検体群で設定した両群を判別する関数(0=5.16x+y+48.11)を用いて胆道がん検出の的中率を算出したところ、真陽性33例、真陰性48例、偽陽性2例、偽陰性0例であり、これらの値から検出性能として精度98%、感度100%、特異度96%が得られた。このようにして、新規に見出された配列番号1~125で表される塩基配列からなるポリヌクレオチドの発現量測定値のいずれか2個の組み合わせ全通りの検出性能を算出した。このうち例として、配列番号1で表される塩基配列からなるポリヌクレオチドと他の配列番号で表される塩基配列からなるポリヌクレオチドとの組み合わせ124通りとその検出性能について表6に記載した。例えば配列番号1及び配列番号7、配列番号1及び配列番号9、配列番号1及び配列番号25、配列番号1及び配列番号66、で表される塩基配列からなるポリヌクレオチドの発現量測定値の組み合わせについても全て、テスト検体群において感度100%を示した。このように既存マーカーであるCA19-9の感度(表5より75.8%)を上回るポリヌクレオチドの発現量測定値の組み合わせはテスト検体群で6,316通り得られ、この組み合わせには実施例1で得られた表2に記載の塩基配列1~125の全てが少なくとも1回は使用された。すなわち、テスト検体群において、配列番号1~125で表される塩基配列からなるポリヌクレオチドの発現量測定値のいずれか2個の組み合わせは、CA19-9を上回る感度で胆道がんを判別することが証明できた。
<全検体を用いた場合の遺伝子マーカーの選定と獲られた遺伝子マーカーの胆道がん判別性能の評価方法>
本実施例では、上記実施例1及び実施例2で用いた学習検体群及びテスト検体群の検体を統合し全検体を用いて、遺伝子マーカーの選定及びその胆道がん判別性能評価を行った。
<テスト検体群検体を用いた複数の遺伝子マーカーの組合せによる胆道がん特異的な判別性能の評価方法>
本実施例では、参考例2に記載した検体群の学習検体群を対象として、実施例1に記載の方法と同様の方法で、胆道がん患者と健康人、大腸がん患者、胃がん患者、食道がん患者、肝がん患者及び膵胆道良性疾患患者からなる対照群との血清中のmiRNAの遺伝子発現量の比較を行い、追加の診断用遺伝子マーカーを選択した。その結果選択された、追加の診断用遺伝子マーカー(配列番号466~478;表1参照)と実施例1で選定された遺伝子マーカーを組み合わせた群から選ばれる1個又は2個以上のマーカーを用いて胆道がん特異的な判別性能を評価した。
<既存血中腫瘍マーカーの胆道がん判別性能>
上記の参考例1で得た学習検体群とテスト検体群について、既存の腫瘍マーカーCEA及びCA19-9の血中濃度を測定した。これらの腫瘍マーカーは、原則、非特許文献2に記載される基準値(CEAは5ng/mL、CA19-9は37U/mL)よりも血中濃度が高いとがんの疑いがあるとされる。従って、各検体毎にCEA及びCA19-9の血中濃度が基準値を超えているか否かを確認し、その結果が胆道がん患者をがんと判定しているか見定め、学習検体群及びテスト検体群における各既存マーカーの感度を算出した。この結果を表5に示した。学習検体群においてはCEAの感度は31.3%、CA19-9の感度は68.2%、テスト検体群においてはCEAの感度は33.3%、CA19-9の感度は59.4%しかなく、いずれのマーカーも胆道がんの検出には有用でないことが分かった(表5)。
Claims (19)
- 胆道がんマーカーである、miR-125a-3p、miR-6893-5p、miR-4476、miR-4294、miR-150-3p、miR-6729-5p、miR-7641、miR-6765-3p、miR-6820-5p、miR-575、miR-6836-3p、miR-1469、miR-663a、miR-6075、miR-4634、miR-423-5p、miR-4454、miR-7109-5p、miR-6789-5p、miR-6877-5p、miR-4792、miR-4530、miR-7975、miR-6724-5p、miR-8073、miR-7977、miR-1231、miR-6799-5p、miR-615-5p、miR-4450、miR-6726-5p、miR-6875-5p、miR-4734、miR-16-5p、miR-602、miR-4651、miR-8069、miR-1238-5p、miR-6880-5p、miR-8072、miR-4723-5p、miR-4732-5p、miR-6125、miR-6090、miR-7114-5p、miR-564、miR-451a、miR-3135b、miR-4497、miR-4665-5p、miR-3622a-5p、miR-6850-5p、miR-6821-5p、miR-5100、miR-6872-3p、miR-4433-3p、miR-1227-5p、miR-3188、miR-7704、miR-3185、miR-1908-3p、miR-6781-5p、miR-6805-5p、miR-8089、miR-665、miR-4486、miR-6722-3p、miR-1260a、miR-4707-5p、miR-6741-5p、miR-1260b、miR-1246、miR-6845-5p、miR-4638-5p、miR-6085、miR-1228-3p、miR-4534、miR-5585-3p、miR-4741、miR-4433b-3p、miR-197-5p、miR-718、miR-4513、miR-4446-3p、miR-619-5p、miR-6816-5p、miR-6778-5p、miR-24-3p、miR-1915-3p、miR-4665-3p、miR-4449、miR-6889-5p、miR-486-3p、miR-7113-3p、miR-642a-3p、miR-7847-3p、miR-6768-5p、miR-1290、miR-7108-5p、miR-92b-5p、miR-663b、miR-3940-5p、miR-4467、miR-6858-5p、miR-4417、miR-3665、miR-4736、miR-4687-3p、miR-1908-5p、miR-5195-3p、miR-4286、miR-3679-3p、miR-6791-5p、miR-1202、miR-3656、miR-4746-3p、miR-3184-5p、miR-3937、miR-6515-3p、miR-6132、miR-187-5p、miR-7111-5p、miR-5787、miR-6779-5p、miR-4516、miR-4649-5p、miR-760、miR-3162-5p、miR-3178、miR-940、miR-4271、miR-6769b-5p、miR-4508、miR-6826-5p、miR-6757-5p、miR-3131、及び、miR-1343-3pからなる群から選択される少なくとも1つ以上のポリヌクレオチドと特異的に結合可能な核酸を含む、胆道がんの検出用キット。
- miR-125a-3pがhsa-miR-125a-3pであり、miR-6893-5pがhsa-miR-6893-5pであり、miR-4476がhsa-miR-4476であり、miR-4294がhsa-miR-4294であり、miR-150-3pがhsa-miR-150-3pであり、miR-6729-5pがhsa-miR-6729-5pであり、miR-7641がhsa-miR-7641であり、miR-6765-3pがhsa-miR-6765-3pであり、miR-6820-5pがhsa-miR-6820-5pであり、miR-575がhsa-miR-575であり、miR-6836-3pがhsa-miR-6836-3pであり、miR-1469がhsa-miR-1469であり、miR-663aがhsa-miR-663aであり、miR-6075がhsa-miR-6075であり、miR-4634がhsa-miR-4634であり、miR-423-5pがhsa-miR-423-5pであり、miR-4454がhsa-miR-4454であり、miR-7109-5pがhsa-miR-7109-5pであり、miR-6789-5pがhsa-miR-6789-5pであり、miR-6877-5pがhsa-miR-6877-5pであり、miR-4792がhsa-miR-4792であり、miR-4530がhsa-miR-4530であり、miR-7975がhsa-miR-7975であり、miR-6724-5pがhsa-miR-6724-5pであり、miR-8073がhsa-miR-8073であり、miR-7977がhsa-miR-7977であり、miR-1231がhsa-miR-1231であり、miR-6799-5pがhsa-miR-6799-5pであり、miR-615-5pがhsa-miR-615-5pであり、miR-4450がhsa-miR-4450であり、miR-6726-5pがhsa-miR-6726-5pであり、miR-6875-5pがhsa-miR-6875-5pであり、miR-4734がhsa-miR-4734であり、miR-16-5pがhsa-miR-16-5pであり、miR-602がhsa-miR-602であり、miR-4651がhsa-miR-4651であり、miR-8069がhsa-miR-8069であり、miR-1238-5pがhsa-miR-1238-5pであり、miR-6880-5pがhsa-miR-6880-5pであり、miR-8072がhsa-miR-8072であり、miR-4723-5pがhsa-miR-4723-5pであり、miR-4732-5pがhsa-miR-4732-5pであり、miR-6125がhsa-miR-6125であり、miR-6090がhsa-miR-6090であり、miR-7114-5pがhsa-miR-7114-5pであり、miR-564がhsa-miR-564であり、miR-451aがhsa-miR-451aであり、miR-3135bがhsa-miR-3135bであり、miR-4497がhsa-miR-4497であり、miR-4665-5pがhsa-miR-4665-5pであり、miR-3622a-5pがhsa-miR-3622a-5pであり、miR-6850-5pがhsa-miR-6850-5pであり、miR-6821-5pがhsa-miR-6821-5pであり、miR-5100がhsa-miR-5100であり、miR-6872-3pがhsa-miR-6872-3pであり、miR-4433-3pがhsa-miR-4433-3pであり、miR-1227-5pがhsa-miR-1227-5pであり、miR-3188がhsa-miR-3188であり、miR-7704がhsa-miR-7704であり、miR-3185がhsa-miR-3185であり、miR-1908-3pがhsa-miR-1908-3pであり、miR-6781-5pがhsa-miR-6781-5pであり、miR-6805-5pがhsa-miR-6805-5pであり、miR-8089がhsa-miR-8089であり、miR-665がhsa-miR-665であり、miR-4486がhsa-miR-4486であり、miR-6722-3pがhsa-miR-6722-3pであり、miR-1260aがhsa-miR-1260aであり、miR-4707-5pがhsa-miR-4707-5pであり、miR-6741-5pがhsa-miR-6741-5pであり、miR-1260bがhsa-miR-1260bであり、miR-1246がhsa-miR-1246であり、miR-6845-5pがhsa-miR-6845-5pであり、miR-4638-5pがhsa-miR-4638-5pであり、miR-6085がhsa-miR-6085であり、miR-1228-3pがhsa-miR-1228-3pであり、miR-4534がhsa-miR-4534であり、miR-5585-3pがhsa-miR-5585-3pであり、miR-4741がhsa-miR-4741であり、miR-4433b-3pがhsa-miR-4433b-3pであり、miR-197-5pがhsa-miR-197-5pであり、miR-718がhsa-miR-718であり、miR-4513がhsa-miR-4513であり、miR-4446-3pがhsa-miR-4446-3pであり、miR-619-5pがhsa-miR-619-5pであり、miR-6816-5pがhsa-miR-6816-5pであり、miR-6778-5pがhsa-miR-6778-5pであり、miR-24-3pがhsa-miR-24-3pであり、miR-1915-3pがhsa-miR-1915-3pであり、miR-4665-3pがhsa-miR-4665-3pであり、miR-4449がhsa-miR-4449であり、miR-6889-5pがhsa-miR-6889-5pであり、miR-486-3pがhsa-miR-486-3pであり、miR-7113-3pがhsa-miR-7113-3pであり、miR-642a-3pがhsa-miR-642a-3pであり、miR-7847-3pがhsa-miR-7847-3pであり、miR-6768-5pがhsa-miR-6768-5pであり、miR-1290がhsa-miR-1290であり、miR-7108-5pがhsa-miR-7108-5pであり、miR-92b-5pがhsa-miR-92b-5pであり、miR-663bがhsa-miR-663bであり、miR-3940-5pがhsa-miR-3940-5pであり、miR-4467がhsa-miR-4467であり、miR-6858-5pがhsa-miR-6858-5pであり、miR-4417がhsa-miR-4417であり、miR-3665がhsa-miR-3665であり、miR-4736がhsa-miR-4736であり、miR-4687-3pがhsa-miR-4687-3pであり、miR-1908-5pがhsa-miR-1908-5pであり、miR-5195-3pがhsa-miR-5195-3pであり、miR-4286がhsa-miR-4286であり、miR-3679-3pがhsa-miR-3679-3pであり、miR-6791-5pがhsa-miR-6791-5pであり、miR-1202がhsa-miR-1202であり、miR-3656がhsa-miR-3656であり、miR-4746-3pがhsa-miR-4746-3pであり、miR-3184-5pがhsa-miR-3184-5pであり、miR-3937がhsa-miR-3937であり、miR-6515-3pがhsa-miR-6515-3pであり、miR-6132がhsa-miR-6132であり、miR-187-5pがhsa-miR-187-5pであり、miR-7111-5pがhsa-miR-7111-5pであり、miR-5787がhsa-miR-5787であり、miR-6779-5pがhsa-miR-6779-5pであり、miR-4516がhsa-miR-4516であり、miR-4649-5pがhsa-miR-4649-5pであり、miR-760がhsa-miR-760であり、miR-3162-5pがhsa-miR-3162-5pであり、miR-3178がhsa-miR-3178であり、miR-940がhsa-miR-940であり、miR-4271がhsa-miR-4271であり、miR-6769b-5pがhsa-miR-6769b-5pであり、miR-4508がhsa-miR-4508であり、miR-6826-5pがhsa-miR-6826-5pであり、miR-6757-5pがhsa-miR-6757-5pであり、miR-3131がhsa-miR-3131であり、及び、miR-1343-3pがhsa-miR-1343-3pである、請求項1に記載のキット。
- 前記核酸が、下記の(a)~(e)に示すポリヌクレオチド:
(a)配列番号1、2、4~125、及び466~478のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(b)配列番号1、2、4~125、及び466~478のいずれかで表される塩基配列を含むポリヌクレオチド、
(c)配列番号1、2、4~125、及び466~478のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(d)配列番号1、2、4~125、及び466~478のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列に相補的な塩基配列を含むポリヌクレオチド、及び
(e)前記(a)~(d)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド、
からなる群から選択されるポリヌクレオチドである、請求項1又は2に記載のキット。 - 前記キットが、別の胆道がんマーカーである、miR-6808-5p、miR-6774-5p、miR-4656、miR-6806-5p、miR-1233-5p、miR-328-5p、miR-4674、miR-2110、miR-6076、miR-3619-3p、miR-92a-2-5p、miR-128-1-5p、miR-638、miR-2861、miR-371a-5p、miR-211-3p、miR-1273g-3p、miR-1203、miR-122-5p、miR-4258、miR-4484、miR-4648及びmiR-6780b-5pからなる群から選択される少なくとも1つ以上のポリヌクレオチドと特異的に結合可能な核酸をさらに含む、請求項1~3のいずれかに記載のキット。
- miR-6808-5pがhsa-miR-6808-5pであり、miR-6774-5pがhsa-miR-6774-5pであり、miR-4656がhsa-miR-4656であり、miR-6806-5pがhsa-miR-6806-5pであり、miR-1233-5pがhsa-miR-1233-5pであり、miR-328-5pがhsa-miR-328-5pであり、miR-4674がhsa-miR-4674であり、miR-2110がhsa-miR-2110であり、miR-6076がhsa-miR-6076であり、miR-3619-3pがhsa-miR-3619-3pであり、miR-92a-2-5pがhsa-miR-92a-2-5pであり、miR-128-1-5pがhsa-miR-128-1-5pであり、miR-638がhsa-miR-638であり、miR-2861がhsa-miR-2861であり、miR-371a-5pがhsa-miR-371a-5pであり、miR-211-3pがhsa-miR-211-3pであり、miR-1273g-3pがhsa-miR-1273g-3pであり、miR-1203がhsa-miR-1203であり、miR-122-5pがhsa-miR-122-5pであり、miR-4258がhsa-miR-4258であり、miR-4484がhsa-miR-4484であり、miR-4648がhsa-miR-4648であり、及び、miR-6780b-5pがhsa-miR-6780b-5pである、請求項4に記載のキット。
- 前記核酸が、下記の(f)~(j)に示すポリヌクレオチド:
(f)配列番号126~148のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(g)配列番号126~148のいずれかで表される塩基配列を含むポリヌクレオチド、
(h)配列番号126~148のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(i)配列番号126~148のいずれかで表される塩基配列もしくは当該塩基配列においてuがtである塩基配列に相補的な塩基配列を含むポリヌクレオチド、及び
(j)前記(f)~(i)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド、
からなる群から選択されるポリヌクレオチドである、請求項4又は5に記載のキット。 - 前記キットが、請求項1又は2に記載のすべての胆道がんマーカーから選択される少なくとも2つ以上のポリヌクレオチドのそれぞれと特異的に結合可能な少なくとも2つ以上の核酸を含む、請求項1~6のいずれかに記載のキット。
- 胆道がんマーカーである、miR-125a-3p、miR-6893-5p、miR-4476、miR-4294、miR-150-3p、miR-6729-5p、miR-7641、miR-6765-3p、miR-6820-5p、miR-575、miR-6836-3p、miR-1469、miR-663a、miR-6075、miR-4634、miR-423-5p、miR-4454、miR-7109-5p、miR-6789-5p、miR-6877-5p、miR-4792、miR-4530、miR-7975、miR-6724-5p、miR-8073、miR-7977、miR-1231、miR-6799-5p、miR-615-5p、miR-4450、miR-6726-5p、miR-6875-5p、miR-4734、miR-16-5p、miR-602、miR-4651、miR-8069、miR-1238-5p、miR-6880-5p、miR-8072、miR-4723-5p、miR-4732-5p、miR-6125、miR-6090、miR-7114-5p、miR-564、miR-451a、miR-3135b、miR-4497、miR-4665-5p、miR-3622a-5p、miR-6850-5p、miR-6821-5p、miR-5100、miR-6872-3p、miR-4433-3p、miR-1227-5p、miR-3188、miR-7704、miR-3185、miR-1908-3p、miR-6781-5p、miR-6805-5p、miR-8089、miR-665、miR-4486、miR-6722-3p、miR-1260a、miR-4707-5p、miR-6741-5p、miR-1260b、miR-1246、miR-6845-5p、miR-4638-5p、miR-6085、miR-1228-3p、miR-4534、miR-5585-3p、miR-4741、miR-4433b-3p、miR-197-5p、miR-718、miR-4513、miR-4446-3p、miR-619-5p、miR-6816-5p、miR-6778-5p、miR-24-3p、miR-1915-3p、miR-4665-3p、miR-4449、miR-6889-5p、miR-486-3p、miR-7113-3p、miR-642a-3p、miR-7847-3p、miR-6768-5p、miR-1290、miR-7108-5p、miR-92b-5p、miR-663b、miR-3940-5p、miR-4467、miR-6858-5p、miR-4417、miR-3665、miR-4736、miR-4687-3p、miR-1908-5p、miR-5195-3p、miR-4286、miR-3679-3p、miR-6791-5p、miR-1202、miR-3656、miR-4746-3p、miR-3184-5p、miR-3937、miR-6515-3p、miR-6132、miR-187-5p、miR-7111-5p、miR-5787、miR-6779-5p、miR-4516、miR-4649-5p、miR-760、miR-3162-5p、miR-3178、miR-940、miR-4271、miR-6769b-5p、miR-4508、miR-6826-5p、miR-6757-5p、miR-3131、及び、miR-1343-3pからなる群から選択される少なくとも1つ以上のポリヌクレオチドと特異的に結合可能な核酸を含む、胆道がんの検出用デバイス。
- miR-125a-3pがhsa-miR-125a-3pであり、miR-6893-5pがhsa-miR-6893-5pであり、miR-4476がhsa-miR-4476であり、miR-4294がhsa-miR-4294であり、miR-150-3pがhsa-miR-150-3pであり、miR-6729-5pがhsa-miR-6729-5pであり、miR-7641がhsa-miR-7641であり、miR-6765-3pがhsa-miR-6765-3pであり、miR-6820-5pがhsa-miR-6820-5pであり、miR-575がhsa-miR-575であり、miR-6836-3pがhsa-miR-6836-3pであり、miR-1469がhsa-miR-1469であり、miR-663aがhsa-miR-663aであり、miR-6075がhsa-miR-6075であり、miR-4634がhsa-miR-4634であり、miR-423-5pがhsa-miR-423-5pであり、miR-4454がhsa-miR-4454であり、miR-7109-5pがhsa-miR-7109-5pであり、miR-6789-5pがhsa-miR-6789-5pであり、miR-6877-5pがhsa-miR-6877-5pであり、miR-4792がhsa-miR-4792であり、miR-4530がhsa-miR-4530であり、miR-7975がhsa-miR-7975であり、miR-6724-5pがhsa-miR-6724-5pであり、miR-8073がhsa-miR-8073であり、miR-7977がhsa-miR-7977であり、miR-1231がhsa-miR-1231であり、miR-6799-5pがhsa-miR-6799-5pであり、miR-615-5pがhsa-miR-615-5pであり、miR-4450がhsa-miR-4450であり、miR-6726-5pがhsa-miR-6726-5pであり、miR-6875-5pがhsa-miR-6875-5pであり、miR-4734がhsa-miR-4734であり、miR-16-5pがhsa-miR-16-5pであり、miR-602がhsa-miR-602であり、miR-4651がhsa-miR-4651であり、miR-8069がhsa-miR-8069であり、miR-1238-5pがhsa-miR-1238-5pであり、miR-6880-5pがhsa-miR-6880-5pであり、miR-8072がhsa-miR-8072であり、miR-4723-5pがhsa-miR-4723-5pであり、miR-4732-5pがhsa-miR-4732-5pであり、miR-6125がhsa-miR-6125であり、miR-6090がhsa-miR-6090であり、miR-7114-5pがhsa-miR-7114-5pであり、miR-564がhsa-miR-564であり、miR-451aがhsa-miR-451aであり、miR-3135bがhsa-miR-3135bであり、miR-4497がhsa-miR-4497であり、miR-4665-5pがhsa-miR-4665-5pであり、miR-3622a-5pがhsa-miR-3622a-5pであり、miR-6850-5pがhsa-miR-6850-5pであり、miR-6821-5pがhsa-miR-6821-5pであり、miR-5100がhsa-miR-5100であり、miR-6872-3pがhsa-miR-6872-3pであり、miR-4433-3pがhsa-miR-4433-3pであり、miR-1227-5pがhsa-miR-1227-5pであり、miR-3188がhsa-miR-3188であり、miR-7704がhsa-miR-7704であり、miR-3185がhsa-miR-3185であり、miR-1908-3pがhsa-miR-1908-3pであり、miR-6781-5pがhsa-miR-6781-5pであり、miR-6805-5pがhsa-miR-6805-5pであり、miR-8089がhsa-miR-8089であり、miR-665がhsa-miR-665であり、miR-4486がhsa-miR-4486であり、miR-6722-3pがhsa-miR-6722-3pであり、miR-1260aがhsa-miR-1260aであり、miR-4707-5pがhsa-miR-4707-5pであり、miR-6741-5pがhsa-miR-6741-5pであり、miR-1260bがhsa-miR-1260bであり、miR-1246がhsa-miR-1246であり、miR-6845-5pがhsa-miR-6845-5pであり、miR-4638-5pがhsa-miR-4638-5pであり、miR-6085がhsa-miR-6085であり、miR-1228-3pがhsa-miR-1228-3pであり、miR-4534がhsa-miR-4534であり、miR-5585-3pがhsa-miR-5585-3pであり、miR-4741がhsa-miR-4741であり、miR-4433b-3pがhsa-miR-4433b-3pであり、miR-197-5pがhsa-miR-197-5pであり、miR-718がhsa-miR-718であり、miR-4513がhsa-miR-4513であり、miR-4446-3pがhsa-miR-4446-3pであり、miR-619-5pがhsa-miR-619-5pであり、miR-6816-5pがhsa-miR-6816-5pであり、miR-6778-5pがhsa-miR-6778-5pであり、miR-24-3pがhsa-miR-24-3pであり、miR-1915-3pがhsa-miR-1915-3pであり、miR-4665-3pがhsa-miR-4665-3pであり、miR-4449がhsa-miR-4449であり、miR-6889-5pがhsa-miR-6889-5pであり、miR-486-3pがhsa-miR-486-3pであり、miR-7113-3pがhsa-miR-7113-3pであり、miR-642a-3pがhsa-miR-642a-3pであり、miR-7847-3pがhsa-miR-7847-3pであり、miR-6768-5pがhsa-miR-6768-5pであり、miR-1290がhsa-miR-1290であり、miR-7108-5pがhsa-miR-7108-5pであり、miR-92b-5pがhsa-miR-92b-5pであり、miR-663bがhsa-miR-663bであり、miR-3940-5pがhsa-miR-3940-5pであり、miR-4467がhsa-miR-4467であり、miR-6858-5pがhsa-miR-6858-5pであり、miR-4417がhsa-miR-4417であり、miR-3665がhsa-miR-3665であり、miR-4736がhsa-miR-4736であり、miR-4687-3pがhsa-miR-4687-3pであり、miR-1908-5pがhsa-miR-1908-5pであり、miR-5195-3pがhsa-miR-5195-3pであり、miR-4286がhsa-miR-4286であり、miR-3679-3pがhsa-miR-3679-3pであり、miR-6791-5pがhsa-miR-6791-5pであり、miR-1202がhsa-miR-1202であり、miR-3656がhsa-miR-3656であり、miR-4746-3pがhsa-miR-4746-3pであり、miR-3184-5pがhsa-miR-3184-5pであり、miR-3937がhsa-miR-3937であり、miR-6515-3pがhsa-miR-6515-3pであり、miR-6132がhsa-miR-6132であり、miR-187-5pがhsa-miR-187-5pであり、miR-7111-5pがhsa-miR-7111-5pであり、miR-5787がhsa-miR-5787であり、及び、miR-6779-5pがhsa-miR-6779-5pであり、miR-4516がhsa-miR-4516であり、miR-4649-5pがhsa-miR-4649-5pであり、miR-760がhsa-miR-760であり、miR-3162-5pがhsa-miR-3162-5pであり、miR-3178がhsa-miR-3178であり、miR-940がhsa-miR-940であり、miR-4271がhsa-miR-4271であり、miR-6769b-5pがhsa-miR-6769b-5pであり、miR-4508がhsa-miR-4508であり、miR-6826-5pがhsa-miR-6826-5pであり、miR-6757-5pがhsa-miR-6757-5pであり、miR-3131がhsa-miR-3131であり、及び、miR-1343-3pがhsa-miR-1343-3pである、請求項8に記載のデバイス。
- 前記核酸が、下記の(a)~(e)に示すポリヌクレオチド:
(a)配列番号1、2、4~125、及び466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(b)配列番号1、2、4~125、及び466~478のいずれかで表される塩基配列を含むポリヌクレオチド、
(c)配列番号1、2、4~125、及び466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(d)配列番号1、2、4~125、及び466~478のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列を含むポリヌクレオチド、及び
(e)前記(a)~(d)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド、
からなる群から選択されるポリヌクレオチドである、請求項8又は9に記載のデバイス。 - 前記デバイスが、別の胆道がんマーカーである、miR-6808-5p、miR-6774-5p、miR-4656、miR-6806-5p、miR-1233-5p、miR-328-5p、miR-4674、miR-2110、miR-6076、miR-3619-3p、miR-92a-2-5p、miR-128-1-5p、miR-638、miR-2861、miR-371a-5p、miR-211-3p、miR-1273g-3p、miR-1203、miR-122-5p、miR-4258、miR-4484、miR-4648及びmiR-6780b-5pからなる群から選択される少なくとも1つ以上のポリヌクレオチドと特異的に結合可能な核酸をさらに含む、請求項8~10のいずれかに記載のデバイス。
- miR-6808-5pがhsa-miR-6808-5pであり、miR-6774-5pがhsa-miR-6774-5pであり、miR-4656がhsa-miR-4656であり、miR-6806-5pがhsa-miR-6806-5pであり、miR-1233-5pがhsa-miR-1233-5pであり、miR-328-5pがhsa-miR-328-5pであり、miR-4674がhsa-miR-4674であり、miR-2110がhsa-miR-2110であり、miR-6076がhsa-miR-6076であり、miR-3619-3pがhsa-miR-3619-3pであり、miR-92a-2-5pがhsa-miR-92a-2-5pであり、miR-128-1-5pがhsa-miR-128-1-5pであり、miR-638がhsa-miR-638であり、miR-2861がhsa-miR-2861であり、miR-371a-5pがhsa-miR-371a-5pであり、miR-211-3pがhsa-miR-211-3pであり、miR-1273g-3pがhsa-miR-1273g-3pであり、miR-1203がhsa-miR-1203であり、miR-122-5pがhsa-miR-122-5pであり、miR-4258がhsa-miR-4258であり、miR-4484がhsa-miR-4484であり、miR-4648がhsa-miR-4648であり、及び、miR-6780b-5pがhsa-miR-6780b-5pである、請求項11に記載のデバイス。
- 前記核酸が、下記の(f)~(j)に示すポリヌクレオチド:
(f)配列番号126~148のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(g)配列番号126~148のいずれかで表される塩基配列を含むポリヌクレオチド、
(h)配列番号126~148のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列からなるポリヌクレオチド、その変異体、その誘導体、又は15以上の連続した塩基を含むその断片、
(i)配列番号126~148のいずれかで表される塩基配列、もしくは当該塩基配列においてuがtである塩基配列、に相補的な塩基配列を含むポリヌクレオチド、及び
(j)前記(f)~(i)のいずれかのポリヌクレオチドとストリンジェントな条件でハイブリダイズするポリヌクレオチド、
からなる群から選択されるポリヌクレオチドである、請求項11又は12に記載のデバイス。 - 前記デバイスが、ハイブリダイゼーション技術による測定のためのデバイスである、請求項8~13のいずれかに記載のデバイス。
- 前記ハイブリダイゼーション技術が、核酸アレイ技術である、請求項14に記載のデバイス。
- 前記デバイスが、請求項8又は9に記載のすべての胆道がんマーカーから選択される少なくとも2つ以上のポリヌクレオチドのそれぞれと特異的に結合可能な少なくとも2つ以上の核酸を含む、請求項8~15のいずれかに記載のデバイス。
- 請求項1~7のいずれかに記載のキット又は請求項8~16のいずれかに記載のデバイスを用いて、被験体の検体における標的核酸の発現量を測定し、該測定された発現量と、同様に測定された健常体の対照発現量とを用いて、被験体が胆道がんに罹患していること、又は胆道がんに罹患していないことをin vitroで評価することを含む、胆道がんの検出方法。
- 前記被験体が、ヒトである、請求項17に記載の方法。
- 前記検体が、血液、血清又は血漿である、請求項17又は18に記載の方法。
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KR20230014872A (ko) | 2023-01-30 |
CA2951008A1 (en) | 2015-12-17 |
CN112029863A (zh) | 2020-12-04 |
KR102560983B1 (ko) | 2023-07-28 |
US20170107581A1 (en) | 2017-04-20 |
JPWO2015190542A1 (ja) | 2017-04-20 |
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US20230091956A1 (en) | 2023-03-23 |
KR102490804B1 (ko) | 2023-01-20 |
EP3156500A1 (en) | 2017-04-19 |
CN106414774B (zh) | 2020-10-09 |
RU2017100015A (ru) | 2018-07-12 |
JP2024063166A (ja) | 2024-05-10 |
JP6927701B2 (ja) | 2021-09-01 |
US10633708B2 (en) | 2020-04-28 |
CN106414774A (zh) | 2017-02-15 |
EP3156500A4 (en) | 2018-04-04 |
JP2021191270A (ja) | 2021-12-16 |
JP2023052321A (ja) | 2023-04-11 |
KR20170015509A (ko) | 2017-02-08 |
KR20230113855A (ko) | 2023-08-01 |
KR20220013463A (ko) | 2022-02-04 |
US20200216914A1 (en) | 2020-07-09 |
KR102355758B1 (ko) | 2022-01-26 |
JP7215694B2 (ja) | 2023-01-31 |
BR112016028944A2 (ja) | 2018-01-09 |
JP7454823B2 (ja) | 2024-03-25 |
US11499198B2 (en) | 2022-11-15 |
US11761046B2 (en) | 2023-09-19 |
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