KR20150140597A - Pharmaceutical composition for preventing and treating cancer comprising double-stranded microRNAs as active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating cancer comprising at least one selected from the group consisting of miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608, and a mimetic thereof as an active ingredient. The miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608, or a mimetic thereof of the present invention having effects in promoting apoptosis and inhibiting cell growth and, particularly, having cancer-specific effects, and having properties without effects on normal cells, can be broadly used for effective anticancer treatment without side effects.

Description

A composition for preventing or treating cancer comprising double-stranded microRNAs as an active ingredient {Pharmaceutical composition for preventing and treating cancer comprising double-stranded microRNAs as active ingredient}

The present invention comprises as an active ingredient at least one selected from the group consisting of miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 and mimics thereof, or It relates to a therapeutic pharmaceutical composition.

Technology that suppresses the expression of a specific gene becomes an important tool for the development of therapeutic agents and target verification for disease treatment. Conventionally, a technology for introducing a transgene for a specific gene has been used to suppress a specific gene, and this technology is a method of introducing a transgene in the antisense direction based on a promoter (see: Sheehy, et al. ., Proc. Natl. Acad. Sci., USA, 85:8805-8808, 1988; Smith, et al., Nature, 334:724-726, 1988) and the transgene in the forward direction (sense) based on the promoter. By applying the method of introduction, the scope of its application is expanding (see Napoli, et al., Plant Cell, 2:279-289, 1990; van der Krol, et al., Plant Cell, 2:291). -299, 1990; U.S. Patent No. 5,034,323; U.S. Patent No. 5,231,020; and, U.S. Patent No. 5,283,184).

On the other hand, microRNA (miRNA) is a short single-stranded ribonucleic acid (RNA) having a length of 19 to 25 nucleotides, and is known to be expressed in cells to regulate the expression of many genes (see: Bartel DP, Cell , 116(2):281-297, 2004; He L. and Hannon GJ, Nat. Rev. Genet., 5(7):522-531, 2004). Up to now, about 2,000 species have been discovered in humans.Since one miRNA can regulate several types of mRNA, it is predicted that about 30% of human genes will be regulated by miRNA (Bentwich, et al. ., Nat. Rev. Genet., 37(7):766-770, 2005; Berezikov, et al., Cell, 120(1):21-24, 2005; Lewis, et al., Cell, 120(1 ):15-20, 2005; Krek, et al., Nat. Genet., 37:495-500, 2005; Friedman, et al., Genome Res., 19:92-105, 2009).

miRNAs are naturally made in cells, and most of them are transcribed into primary miRNA (primary miRNA) by RNA polymerase II, and then RNase III enzymes, drosha and pasha, in the nucleus. DGCR8) or the like is cleaved into precursor miRNA (precursor miRNA). Thereafter, the precursor miRNA is transferred from the nucleus to the cytoplasm by exportin-5, and is prepared as a short and incomplete double-stranded miRNA by a dicer, an RNase III enzyme, in the cytoplasm. The prepared miRNA is separated into a single strand by binding to a RISC (RNA-induced silencing complex) protein complex, and the isolated single-stranded miRNA is complementary to the nucleotide sequence of the 3'-UTR (untranslated region) of the target mRNA. It is known to inhibit the expression of genes by binding to the ribosome to inhibit the process of translating mRNA into a protein by the ribosome or by causing the target mRNA to be degraded (Lee, et al., Nature, 425(6956)). :415-419, 2003; Gregory, et al., Nature, 432(7014):235-240, 2004; Yi, et al., Genes & Dev., 17:3011-3016, 2003; Bohnsack, et al. , RNA, 10:185-191, 2004; Hutvagner, et al., Science, 293:834-838, 2001; Ketting, et al., Genes & Dev., 15:2654-2659, 2001; Pillai et al. , Trends Cell Biol., 17:118-126, 2007).

Recently, the expression of specific miRNAs has been increased or decreased in many types of cancer, so miRNAs are expected to be strongly associated with cancer pathogenesis, and by continuous research, various types of miRNAs are cancer suppressors. Or it has been reported that it acts as an oncogene (see: Iorio, et al., Cancer Res., 65(16):7065-7070, 2005; Yu, et al., DNA Cell Biol., 26( 5):283-292, 2007; Soifer, et al., Mol. Ther., 15(12):2070-2079, 2007). For example, miRNAs that act as cancer suppressors include miR-15, miR-16, let-7 miRNA, and miR-34 family, among which miR-15 and miR-16 are Bcl-2 oncogenes. It regulates its expression and decreases its expression in chronic lymphocytic leukemia (Cimmino, et al., Proc. Natl. Acad. Sci., USA., 102(39):13944-13949, 2005), let-7 miRNA inhibits the activities of oncogenes such as RAS, HGMA2, myc, CDK6, and CDC25 (see Johnson, et al., Cell, 120(5):635-647, 2005; Lee YS). , Dutta A., Genes Dev., 21:1025-1030, 2007; Sampson et al., Cancer Res., 67:9762-9770, 2007; Johnson et al., 67:7713-7722, 2007), miR- The 34 family (miR-34a, miR-34b, miR-34c) is regulated by p53 to regulate the expression of target mRNAs such as E2F3, CDK4, CDK6, CCNE2, BIRC5, DCR3, BCL-2, etc. It is known to play a role as an inhibitor (Tarasov et al., Cell Cycle, 6:1586-1593, 2007; He et al., Nature, 447:1130-1134, 2007; Corney et al., Cancer) Res., 67:8433-8438, 2007; Chang et al., Mol. Cell, 26:745-752, 2007).

In contrast, miRNAs that act as carcinogens have been reported as miRNA-17-92-1 cluster, miR-372 and miR-373, miR-21, and miR-155 (He et al., Nature). , 435:828-833, 2005; O'Donnell et al., Nature, 435:839-843, 2005; Voorhoeve, et al., Cell, 124:1169-1181, 2006; Si et al., Oncogene, 26 :2799-2803, 2007; Meng et al., Gastroenterology, 133:647-658, 2007; Costinean et al., Proc. Natl. Acad. Sci. USA, 103:7024-7029, 2006).

As such, cancer diseases and specific miRNAs are related to each other, and as it has been found that various types of miRNAs act as tumor suppressors or oncogenes, studies to perform anticancer treatment using these miRNAs As a result of actively progressing, it has been reported that miRNA can be used to suppress the expression of a target gene (Cimmino, et al., Proc. Natl. Acad. Sci., USA., 102(39):13944-13949 , 2005; Takamizawa, et al., Cancer Res., 64(11):3753-3756, 2004; Akao, et al., Biol. Pharm. Bull., 29(5):903-906, 2006; Scott, et al., J. Biol. Chem., 282(2):1479-1486, 2007; Chan, et al., Cancer Res., 65(14):6029-6033, 2005; Si, et al., Oncogene , 26(19):2799-803, 2007; Krutzfeldt, et al., Nature, 438(7068):685-689, 2005; Esquela-Kerscher, et al., Cell Cycle, 7:759-764, 2008; Kumar, et al., Proc. Natl. Acad. Sci., USA., 105:3903-3908, 2008; Elmen, et al., Nature, 452:896-899, 2008). In the above-described anticancer treatment method, miRNA acting as a cancer suppressor is introduced into cancer cells by introducing a miRNA mimetic or a miRNA expression vector to increase the expression of the corresponding miRNA, or antisense in the case of miRNA acting as a carcinogen. A method of directly inhibiting the proliferation of cancer cells is known by introducing an oligonucleotide (antisense oligonucleotide) into cancer cells to reduce the expression of the corresponding miRNA.

For example, U.S. Patent Publication No. 2006/0189557 discloses a method of treating cancer cells by introducing let-7 miRNA into cancer cells and inhibiting the activity of RAS oncogenes. International Patent Publication No. WO 06133022 discloses miRNA- A method of treating cancer cells by inhibiting miRNAs in the 17-92-1 cluster is disclosed. Patent Publication No. 2009-0081125 binds to mRNA transcribed from the Met gene, inhibiting the expression of the met gene, and A double-stranded miRNA capable of inhibiting growth and an anticancer agent comprising the same as an active ingredient are disclosed.

Since cancer exhibits a very heterogeneous character that exhibits abnormalities in one or more of several cell signaling pathways, as described above, a therapeutic agent that acts only on one target or one pathway has a very low effect of chemotherapy. In order to overcome, a combination therapy in which several inhibitors acting on different targets are administered together or continuously is required, but the combination therapy has a side effect of killing or damaging not only the patient's cancer cells but also normal cells. , As an active ingredient, a therapeutic agent containing miRNAs acting on only one target or one pathway alone or in combination cannot be expected to have a large anticancer effect. However, since miRNA naturally exists in cells, side effects are expected to be relatively low compared to chemically synthesized therapeutic agents, so there is a constant need to develop anticancer drugs that contain miRNA as an active ingredient and have a combination treatment effect. Emerged.

Accordingly, the present inventors made intensive research efforts to develop an anticancer agent having a complex therapeutic effect using miRNA having an excellent anticancer effect. As a result, miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p And miR-608 was developed to confirm that the mimic has a cell death effect and a cell growth inhibitory effect by complexly inhibiting target genes of various pathways, and the present invention was completed.

One object of the present invention is to include as an active ingredient at least one selected from the group consisting of miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 and mimics thereof To provide a pharmaceutical composition for preventing or treating cancer.

As an aspect for achieving the above object, the present invention provides a miR-28-5p mimic, miR-323-5p mimic, miR-510-5p mimic, miR-552-3p mimic, and It provides one or more miRNA mimics selected from the group consisting of miR-608 mimics.

Specifically, the present invention miR-28-5p miR-28-5p in which RNA having the nucleotide sequence of SEQ ID NO: 33 and RNA having the nucleotide sequence of SEQ ID NO: 34 are bound, RNA having the nucleotide sequence of SEQ ID NO: 237, and SEQ ID NO: 238 MiR-323-5p mimic, sequence of miR-323-5p, in which RNA having the base sequence of SEQ ID NO: 365 and RNA having the base sequence of SEQ ID NO: 366 are combined MiR-552-3p miR-552-3p in which RNA having the nucleotide sequence of SEQ ID NO: 413 and RNA having the nucleotide sequence of SEQ ID NO: 414 are combined, and the RNA having the nucleotide sequence of SEQ ID NO: 467 and the RNA having the nucleotide sequence of SEQ ID NO: 468 Provides a mimic of bound miR-608.

In the present invention, most miRNAs or microRNAs are intervened in introns on chromosomes, which are processed by Drosha or the like after transcription and are converted into a precursor form (precursor-miRNA, precursor form). After this, the nucleus is exited by exportin 5 and converted into a mature miRNA of about 18 to 22 bp in length by Dicer. These mature miRNAs exhibit functions by binding to RISC (RNA induced silencing complex) (Nat Rev Mol Cell Biol 6, 376-785; 2005).

The miRNA of the present invention may exist in a single-stranded or double-stranded form. Mature miRNAs that bind RISCs are primarily single-stranded, but precursor miRNAs can have a partially self-complementary (eg stem- and loop structure) structure that can form primarily a double-stranded portion. Meanwhile, the miRNA mimic of the present invention may also exist in a single-stranded or double-stranded form, and in particular, for the purposes of the present invention, it may be a double-stranded miRNA.

As used herein, the term “miRNA mimic” or “mimetic” refers to a substance used to increase the expression and/or function of miRNA, so that the miRNA mimic may increase, supplement or replace the function of natural miRNA. In particular, the miRNA mimic may be a polynucleotide comprising a mature miRNA sequence, a pre-miRNA or a pri-miRNA sequence. For the purposes of the present invention, the miRNA mimic may be a double-stranded miRNA in consideration of the intracellular stability of the miRNA, and may be separated into a single strand when acting as a mature miRNA as described above.

In addition, miRNA mimics include locked nucleic acids, chemical modifications such as peptide nucleic acids, 2'-O-alkyl (e.g., 2'-O-methyl, 2'-O-methoxy ethyl), 2 Fructose mutations such as'-fluoro and 4'thio mutations and backbone mutations such as one or more thiophosphoric acid, morphonoline or carboxylic acid phosphonone linkages. Certain miRNA mimics are also commercially available.

In the present invention, the structure of the mimic can be any one capable of inhibiting the expression of the target gene by the RNAi effect, and both the blunt end or the cohesive end can be used. As for the adhesive end structure, a structure in which the 3'end side protrudes and a structure in which the 5'end side overhangs are possible. The number of protruding nucleotides is not limited, but may be, for example, 1 to 8 nucleotides. In addition, as described above, in order to prevent rapid degradation by nucleic acid degrading enzymes in vivo and increase in vivo stability, chemical mutations may be introduced by a general method known in the art. For example, the hydroxyl group at the 2'-position of the sugar (ribose ring) is H, OR, R, R'OR, SH, SR, NH ₂ , NHR NR ₂ , N ₃ , CN, F, Cl, Br, I Or the like (where R is alkyl or aryl, preferably an alkyl group having 1 to 6 carbon atoms, and R′ may be alkylene, preferably alkylene having 1 to 6 carbon atoms), or the phosphoric acid backbone is phosphorothioate. , Phosphorodithioate, alkylphosphonate, phosphoroamidate, or boranophosphate.

In the present invention, miRNA mimics include variants having one or more substitutions, insertions, deletions, and combinations thereof, which are functional equivalents having changes that do not reduce miRNA activity.

In the present invention, the miRNA mimic is basically a complete form in which two strands of RNA are paired to form a double strand, that is, a form that is directly synthesized in vitro and then introduced into a cell through transfection, or a plasmid It may be a modified form having a short hairpin structure so that it can be used for transfection using a pre-miRNA vector and a PCR-induced miRNA expression cassette.

The miRNA mimic provided in the present invention is a method of direct chemical synthesis, a method of synthesis using in vitro transcription, a method of cutting long double-stranded RNA synthesized by in vitro transcription using an RNaseIII family enzyme, etc. It can be synthesized by various methods known in the art.

In a specific embodiment of the present invention, miRNA sequences from miRNA sequences registered in miRBase (http://www.mirbase.org), a miRNA sequence database constructed at the University of Manchester, are A double-stranded RNA was prepared to be produced. That is, a double-strand designed to bind to the target nucleotide sequence using a method of linking the phosphodiester bonds forming the skeleton of the DNA structure using β-cyanoethyl phosphoramidite. RNA was synthesized (Sinha, et al., Nucleic Acids Research, 12:4539-4557, 1984). Following the above synthesis, a miRNA library containing a total of 300 miRNAs was prepared by combining the sense and antisense RNA strands to prepare the desired double-stranded miRNAs (SEQ ID NOs: 1 to 600; Table 1).

On the other hand, miRNA mimics can be expressed in cells through methods such as transformation or vector. "Vector" is used to deliver a nucleic acid of interest to the inside of a cell, and includes polynucleotides, polynucleotides, plasmids and viruses bound to ionic or amphoteric compounds, but in the present invention, miRNA mimics are delivered into cells. As long as the purpose can be achieved, it is not limited thereto.

In another aspect, the present invention uses at least one selected from the group consisting of miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608, and mimics thereof as an active ingredient. It provides a pharmaceutical composition for preventing or treating cancer comprising as.

The present invention provides a pharmaceutical composition for preventing or treating lung cancer, liver cancer or colon cancer comprising miR-552-3p or a mimic thereof as an active ingredient. In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer comprising miR-323-5p or a mimic thereof as an active ingredient, and in particular, cancer comprising the miR-323-5p or mimic thereof as an active ingredient The pharmaceutical composition for preventing or treating may be a pharmaceutical composition for preventing or treating uterine cancer, breast cancer, or prostate cancer. In addition, the present invention provides a pharmaceutical composition for preventing or treating lung cancer, prostate cancer, liver cancer, colon cancer or breast cancer, comprising miR-608 or a mimic thereof as an active ingredient.

Specifically, the present invention miR-28-5p miR-28-5p in which RNA having the nucleotide sequence of SEQ ID NO: 33 and RNA having the nucleotide sequence of SEQ ID NO: 34 are bound, RNA having the nucleotide sequence of SEQ ID NO: 237, and SEQ ID NO: 238 MiR-323-5p mimic, sequence of miR-323-5p, in which RNA having the base sequence of SEQ ID NO: 365 and RNA having the base sequence of SEQ ID NO: 366 are combined MiR-552-3p miR-552-3p in which RNA having the nucleotide sequence of SEQ ID NO: 413 and RNA having the nucleotide sequence of SEQ ID NO: 414 are combined, and the RNA having the nucleotide sequence of SEQ ID NO: 467 and the RNA having the nucleotide sequence of SEQ ID NO: 468 It provides a pharmaceutical composition for preventing or treating cancer comprising as an active ingredient at least one mimic selected from the group consisting of miR-608 combined mimics.

The term "mimetic" of the present invention is as described above.

In the present invention, the miRNA or mimics thereof may be included in a complex form with various nucleic acid delivery systems (viral or non-viral delivery systems) known in the art in order to increase delivery efficiency in vivo. For example, it may be included as a recombinant plasmid or viral vector expressing the miRNAs or miRNA mimetics of the present invention. Plasmids that can be used for this are, for example, pSilencer (Ambion), pSiEx (Novagen), siXpress (Takara Bio), pBLOCK-iT ^TM , pcDNA3.1, pCEP4 (Invtrogen), SilenCircle ^TM (Allele), etc. It is not limited thereto. Retrovirus vectors, adenovirus vectors, adeno-associated virus vectors, vaccinia virus vectors, lentiviral vectors, herpes virus vectors, alpha virus vectors, EB virus vectors, papillomavirus vectors, and poomivirus vectors can be used as viral carriers. And is not limited thereto. In addition, as a non-viral carrier, as a delivery reagent, MirusTrasIT-TKO lipophilic reagent, lipofectin, lipofectamine, cellfectin, G-fectin, cationic phospholipid nanoparticles, cationic polymer, cationic micelle, Cationic emulsions or liposomes, ligand-DNA complexes, and gene guns may be used, but are not limited thereto. In the form of liposomes, it is combined with lipids present as amphipathic agents, for example micelles, insoluble monolayers, liquid crystals, or lamellar layers present in aqueous solutions. Lipids for liposome formulations include, but are not limited to, monoglycerides, diglycerides, sulfatides, lysolecithin, lecithin phospholipids, saponins, bile acids, lipofectins, and the like.

In addition, in order to increase the stability of the miRNA of the present invention or its mimic in the body, a biocompatible polymer such as polyethylene glycol is conjugated to increase intracellular absorption. I can. It also includes a method of suspending cells in a solution containing miRNAs or mimics thereof by electroporation and passing them through a high voltage pulse of direct current to be introduced into the cells. In a living body, a solution containing miRNA or mimic is administered to a desired site, and a DC voltage is pulsed through an electrode to be introduced into the cell.

The composition of the present invention can be used both in vitro and in vivo.

In the present invention, the term "cancer treatment or prevention" refers to an action of inhibiting or killing the proliferation of cancer cells and an action of inhibiting or blocking metastasis of cancer cells. In the present invention, "prevention" refers to any action that inhibits cancer formation or delays the onset of cancer by administration of the composition, and "treatment" refers to any action in which the symptoms of the disease are improved or beneficially changed by the administration of the composition. It means.

As used herein, "cancer" refers to gastric cancer, bladder cancer, brain tumor, breast cancer, bone marrow cancer, cervical cancer, chronic lymphocytic leukemia, colon cancer, esophageal cancer, lymphoblastic leukemia, follicular lymphoma, T-cell or B-cell origin. Sheep malignancies, myelogenous leukemia, myeloma, oral cancer, ovarian cancer, prostate cancer, spleen cancer, fibrosarcoma, myoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, lymphangioendopiacytoma , Synovioma, mesothelioma, leiomyosarcoma, rhabdomyosarcoma, colon tumor, rectal cancer, pancreatic cancer, uterine cancer, head and neck cancer, skin cancer, impression cell tumor, sebaceous gland tumor, papillary tumor, papillary adenoma, cystic adenocarcinoma, medullary tumor, organ support Tumor, renal cell tumor, liver cancer, biliary adenoma, chorionic cancer, normal hematoma, fetal tumor, Wilm's tumor, testicular cancer, lung tumor, small cell lung tumor, non-small cell lung tumor, bladder tumor, epithelial tumor, glioma , Astrocytoma, medulloblastoma, cranial cephaloma, ventricular cytoma, pineal gliomas, hemangioblastoma, auditory neuroma, glioblastoma, nasopharyngeal carcinoma, male jaw, melanoma, neuroblastoma, retinoblastoma, leukemia, lymphoma and Kaposi ( Kaposi) sarcoma, etc.

In a specific embodiment of the present invention, the miRNA of the present invention is mixed with Opti-MEM medium, and a solution for transformation is prepared using lipofectamine RNAiMAX, Invitrogen, USA, and then treated with a lung cancer cell line (A549 cell line). I did. As a result of checking the cancer cell growth inhibitory effect, 49 miRNAs inhibited the growth of cancer cells by 70% or less compared to the control group, and 5 miRNAs were 60% or less. In particular, miR-608 showed better cancer cell growth inhibitory effect than miR-16 and miR-542-3p, which were used as positive controls. Five miRNAs (miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608) were selected for cancer cell growth less than 60% with good cancer growth inhibitory effect (Fig. 1). .

In addition, the selected miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, and miR-608 mimics were verified to inhibit cell proliferation in other cancer cell lines other than the A549 lung cancer cell line. To do this, the mimics were transfected into uterine cancer cells HeLa, breast cancer cells MCF7, prostate cancer cells PC3, HeLa contaminated cells KB, lung cancer cells NCI-H460, liver cancer cells Hep3B, colon cancer cells HCT116, and normal cells, fibroblast WI-38. After conversion, the effect of inhibiting cell proliferation was confirmed. As a result, miR-323-5p and miR-608 showed good inhibitory effect on cell growth, and miR-608 and miR-552-3p showed excellent inhibitory effect in lung cancer cells, NCI-H460 and A549. It was confirmed that 552-3p and miR-608 can be usefully used in the treatment of lung cancer. miR-323-5p has a good inhibitory effect on uterine cancer cells HeLa, breast cancer cells MCF7 and prostate cancer cells PC3, so it has been confirmed that it can be usefully used in the treatment of uterine cancer, breast cancer and prostate cancer. It was confirmed that it did not inhibit growth, and it was confirmed that miR-323-5p can be usefully used because of its high specificity for cancer cells with little or no side effects on normal cells (FIG. 2).

By the examples of the present invention, it was confirmed that the miR-552-3p and miR-608 or mimics thereof of the present invention have a specific cancer prevention or treatment effect, and the miR-323-5p or mimics thereof have uterine cancer and breast cancer. Alternatively, the effect of preventing or treating cancer specific to prostate cancer was confirmed. Particularly, in the case of miR-323-5p, it was confirmed that the cancer cell growth inhibitory effect was superior to that of miR-608, but it did not affect the growth of normal cells, so that side effects were small or cancer cell specificity was high.

The composition of the present invention may be for promoting apoptosis or inhibiting cell growth. In a specific embodiment of the present invention, the miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p and miR-608 mimetics of the present invention significantly reduce the cell growth of the lung cancer cell line A549. It was confirmed that it promotes cell death (FIGS. 1 and 5).

In one embodiment of the present invention, as a result of confirming how the miRNA of the present invention affects proteins related to apoptosis and cell cycle, miR-323-5p is Bcl-xL and BIRC5, and miR-510-5p is Mcl-1 and BIRC5, miR-552-3p reduced the protein expression levels of Mcl-1 and BIRC5, and miR-608 reduced Bcl-xL, and miR-28-5p, miR-510-5p, and miR-608 were apoptosis-inducing proteins. By confirming that the protein expression level of BIK is increased, the effect of inducing complex cell death of the miRNAs of the present invention was confirmed. In addition, miR-28-5p is CCND3, miR-323-5p is CDK2, CDK4, CCND1, E2F1, miR-552-3p is CDK6 and CCND3, miR-608 is CDK6, CCND1, CCND3. By confirming this, it was confirmed that the expression of proteins required for cell cycle progression was suppressed by the miRNAs of the present invention. In addition, when the miRNA of the present invention was transformed, it was found that phosphorylation of Ser473 and Thr308, which means Akt activation, was significantly reduced, and by miR-323-5p and miR-608, PI3K, an enzyme required for Akt activation, It was confirmed that the expression level of the regulatory subunit PIK3R2 protein was significantly reduced. In addition, it was confirmed that Sp1, a transcription factor that broadly affects cell differentiation, growth, and apoptosis, also inhibits protein expression by miR-28-5p, miR-323-5p, miR-510-5p, and miR-608. I did. Therefore, when the miRNA of the present invention was treated, it was confirmed that proteins required for cell survival and growth were complexly regulated (Table 4). This confirms the superiority of the miRNAs of the present invention in consideration of the multifactorial and heterogeneous properties of cancer.

In addition, the pharmaceutical composition for preventing or treating cancer of the present invention may further include a pharmaceutically acceptable carrier, and may be formulated together with the carrier.

In the present invention, the term "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not stimulate an organism and does not inhibit the biological activity and properties of the administered compound. Acceptable pharmaceutical carriers for compositions formulated as liquid solutions are sterilized and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as needed. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injection formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

The pharmaceutical composition for preventing or treating cancer comprising the miRNA or mimic thereof of the present invention can be applied in any dosage form containing it as an active ingredient, and can be prepared in an oral or parenteral dosage form. The pharmaceutical formulation of the present invention is oral, rectal, nasal, topical (including cheek and sublingual), subcutaneous, vaginal or parenteral; intramuscular, subcutaneous And those suitable for administration, including intravenous), or in forms suitable for administration by inhalation or insufflation.

Formulations for oral administration comprising the composition of the present invention as an active ingredient include, for example, tablets, troches, lozenges, water-soluble or oily suspensions, powders or granules, emulsions, hard or soft capsules, syrup or elixirs. can do. For formulation into tablets and capsules, etc., binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin, excipients such as dicalcium phosphate, disintegrants such as corn starch or sweet potato starch, and masne stearate Lubricating oils such as calcium, calcium stearate, sodium stearyl fumarate, or polyethylene glycol wax may be included, and in the case of capsule formulation, a liquid carrier such as fatty oil may be further included in addition to the above-mentioned substances.

Formulations for parenteral administration containing the composition of the present invention as an active ingredient include injection forms such as subcutaneous injection, intravenous injection, or intramuscular injection, suppository injection, or spray such as an aerosol that enables inhalation through the respiratory tract. It can be formulated as.

In order to formulate a formulation for injection, the composition of the present invention may be prepared as a solution or suspension by mixing in water together with a stabilizer or buffer, and it may be formulated for unit administration in ampoules or vials. In order to inject as a suppository, it can be formulated into a composition for rectal administration such as a suppository containing a conventional suppository base such as cocoa butter or other glycerides or a body care enema. When formulated for spraying such as an aerosol, a propellant or the like may be blended together with an additive so that the water-dispersed concentrate or wet powder is dispersed.

The composition of the present invention is prepared in the form of a composition for injection so that it can be injected into an area where a tumor occurs or is at risk, or in the form of a gel composition or an adhesive composition for transdermal absorption, so that it can be directly applied or attached to the area where the tumor has occurred It is desirable to do it. At this time, the composition for injection is prepared in the form of an isotonic aqueous solution or suspension to contain an auxiliary agent (e.g., a preservative, a stabilizer, a wetting agent or an emulsifier solution accelerator, a salt for controlling osmotic pressure, a buffer and/or a liposome preparation) The gel composition may contain a gel formulation such as carboxymethyl cellulose, methyl cellulose, acrylic acid polymer, carbopol, and a pharmaceutically acceptable carrier and/or liposome formulation, and the pressure-sensitive adhesive for transdermal absorption The active ingredient layer includes a pressure-sensitive adhesive layer, an adsorption layer for sebum absorption, and a therapeutic drug layer, and the therapeutic drug layer contains a pharmaceutically acceptable carrier and/or a liposome preparation.

In addition, the composition of the present invention further comprises at least one active ingredient selected from the group consisting of known anticancer agents, chemotherapeutic agents, immunotherapy agents, antibacterial agents, radiotherapy agents and antiviral agents, and photodynamic therapy. It can increase the effect.

The amount of miRNA or miRNA thereof contained in the composition of the present invention may be adjusted according to the characteristics and severity of the disease, and characteristics that have been conventionally treated to the patient, and from about 10 micrograms per 1 kg of body or organ weight to It can be made to contain about 20mg. In addition, the dosage depends on the expression vector and the administration target, for example, in the case of a viral vector, the amount of the recombinant virus containing the viral vector is in the range of 10 ³ to 10 ¹² pfu/kg. In particular, miRNA or miRNAs thereof included in the composition of the present invention may be administered 1 to 5 mg per kg of body weight each day, and the dosage level for patients exhibiting specific symptoms is the weight, age, and sex of the patient. , Health condition, diet, administration time, administration method, excretion rate, severity, etc., one of ordinary skill in the art can change the dosage.

In another aspect, the present invention uses at least one selected from the group consisting of miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608, and mimics thereof as an active ingredient. It provides a method of treating cancer comprising administering to an individual a pharmaceutical composition for preventing or treating cancer, including as.

In the present invention, the term "administration" means introducing the pharmaceutical composition of the present invention to a patient by any suitable method, and transport or miRNA of the present miRNA or miRNA of the present invention by viral or non-viral technology Or transplantation of cells expressing their mimics. The route of administration of the composition of the present invention may be administered through various routes, either oral or parenteral, as long as it can reach the target tissue, and specifically, oral, rectal, topical, intravenous, intraperitoneal, intramuscular, intraarterial, It can be administered in a conventional manner via transdermal, intranasal, inhalation, intraocular or intradermal routes. Preferably, it is topically administered to cancer tissues.

The treatment method of the present invention includes administering the anticancer composition of the present invention in a pharmaceutically effective amount. It is obvious to those skilled in the art that the appropriate total daily use amount can be determined by the treating physician within the range of correct medical judgment. A specific therapeutically effective amount for a particular patient is the type and extent of the reaction to be achieved, the specific composition, including whether or not other agents are used in some cases, the patient's age, weight, general health status, sex and diet, administration time, It is preferable to apply differently according to various factors including the route of administration and the secretion rate of the composition, the treatment period, drugs used with or concurrently with the specific composition, and similar factors well known in the field of medicine. Therefore, it is preferable to determine the effective amount of the anticancer composition suitable for the purposes of the present invention in consideration of the above-described matters.

The miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 or their mimics of the present invention have the effect of promoting apoptosis and inhibiting cell growth. It has a cancer-specific effect and does not affect normal cells, so it has no side effects and can be widely used for effective anticancer treatment.

1 is a graph showing the results of comparing the change in the number of cells by transforming 300 miRNA mimics of the present invention into A549 lung cancer cells, treating the CCK-8 (Dojindo Laboratories, Japan) reagent, and measuring the absorbance.
Figure 2 is to verify the cell proliferation inhibitory effect by miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 mimics in various cancer cell lines, cervical cancer cells HeLa , Breast cancer cells MCF7, prostate cancer cells PC3, HeLa contaminated cells KB, lung cancer cells NCI-H460, liver cancer cells Hep3B, colon cancer cells HCT116 and normal cells, fibroblasts WI-38 72 hours after transformation of the mimic It is a diagram comparing the change in the number of cells in each cancer cell line.
3 shows miR-28-5p, miR-323-5p, miR-510-5p, miR-552- in lung cancer cells A549 in order to verify the effect of inhibiting colony formation by miRNA mimics of the present invention. This is a diagram comparing the results of counting the number of colonies formed after 3p, miR-608 mimic was transformed, cultured for 10 days, and stained with crystal violet. At this time, as a control, cancer cells transformed with an NC mimic (negative control mimic) were used. Specifically, Figure 3a compares the number of colonies of lung cancer cell lines treated with miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 mimic and NC mimic, respectively. It is a graph, and Figure 3b is a photograph of colony formation of lung cancer cell lines treated with miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 mimic and NC mimic, respectively to be.
Figure 4 is a cell cycle measured by flow cytometry for lung cancer cell lines treated with miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 mimic and NC mimic, respectively. It is a diagram showing the state of steps.
Figure 5 shows Annexin V staining (Annexin V) for the induction of apoptosis in A549 cells transformed with miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, and miR-608 mimics. -V-FLUOS, Roche Applied Science, Germany) is a picture comparing the results. Specifically, FIG. 5A is the result of the experiment on the 2nd day after transformation, and FIG. 5B is the result of the experiment on the 3rd day after transformation.
6 shows proteins related to cell death and cell cycle targeting A549 cells transformed with miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, and miR-608 mimics. It is a diagram showing the results of Western blotting using antibodies against them. Specifically, Figure 6a is a Western blot photograph showing the levels of proteins related to apoptosis expressed in cells transformed with the double-stranded miRNA of the present invention, Figure 6b is expressed in cells transformed with the double-stranded miRNA of the present invention It is a Western blot photograph showing the protein levels related to the cell cycle, and FIG. 6C is a Western blot photograph showing the protein levels related to the Akt pathway expressed in cells transformed with the double-stranded miRNA of the present invention.
7 is a cell after 72 hours to determine the degree of growth inhibition of lung cancer cells A549 transformed with siRNAs that inhibit the expression of Bcl-xL, BIRC5, CCND1, CCND3, CDK6, CYB5R3, Mcl-1, PIK3R2, Sp1 genes. This is the result of measuring the number.
8 is a human lung cancer cell NCI-H460 transformed with a miR-608 mimic in order to confirm the tumor proliferation inhibitory effect of nude mice by the human lung cancer cell NCI-H460 transformed with miR-608 mimic is transplanted into a mouse. It is a diagram showing the results of measuring the tumor size over time.

Hereinafter, the present invention will be described in more detail by the following examples. However, the following examples are for illustrative purposes only, and the scope of the present invention is not limited thereto.

Example 1: target sequence design and miRNA Mimic Produce

Double-stranded RNA was prepared so that miRNA sequences were generated in cells from miRNA sequences registered in miRBase (http://www.mirbase.org), a miRNA sequence database established at the University of Manchester. .

That is, a double-strand designed to bind to the target nucleotide sequence using a method of linking the phosphodiester bonds forming the skeleton of the DNA structure using β-cyanoethyl phosphoramidite. RNA was synthesized (Sinha, et al., Nucleic Acids Research, 12:4539-4557, 1984). Using an RNA synthesizer (Perseptive Biosystems 8909, PE Biosystems, USA), a series of processes consisting of deblocking, coupling, oxidation and capping are performed on a solid support to which nucleotides are attached. It was repeated to obtain a reaction containing RNA of the desired length. RNA was separated by HPLC LC918 (Japan Analytical Industry, Japan) using Daisogel C18 (Daiso, Japan), and RNA separated by MALDI-TOF mass spectrometry (Shimadzu, Japan) was synthesized with the nucleotide sequence to be synthesized. We checked if they match. Then, by combining the sense and antisense RNA strands, the desired double-stranded miRNAs (SEQ ID NOs: 1 to 600) were prepared.

Example 2: tumor cell culture and miRNA Mimic Transformation using

Human lung cancer cells (A549), uterine cancer cells (HeLa), breast cancer cells (MCF7), prostate cancer cells (PC3), HeLa contaminated cells (KB), lung cancer cells (NCI-) obtained from the Korean Cell Line Bank. H460) and colon cancer cells (HCT116) were cultured in RPMI culture medium (GIBCO/Invitrogen, USA) containing 10% (v/v) fetal calf serum, 100 units/ml penicillin and 100 μg/ml streptomycin, Liver cancer cells (Hep3B) were cultured in DMEM culture medium (GIBCO/Invitrogen, USA) containing 10% (v/v) fetal calf serum, 100 units/ml penicillin and 100 μg/ml streptomycin, and WI -38 (fibroblast) was purchased from the American Type Culture Collection (ATCC), and Eagle's Minimum containing 10% (v/v) fetal calf serum, 100 units/ml penicillin and 100 μg/ml streptomycin. It was cultured in Essential Medium (ATCC, USA). ^{For transformation, 1.5 X 10 5} cells per well of a 6-well plate were dispensed, cultured for 24 hours under conditions of 37° C., 5% (v/v) CO ₂ , and Opti-MEM medium (GIBCO/Invitrogen , USA) After washing once with 2 ml, 500 μl of Opti-MEM medium per well was dispensed.

Transfection reagent for transforming the cell line was mixed with 250 µl Opti-MEM medium so that the double-stranded miRNA prepared in Example 1 was at a final concentration of 20 nM (20 pmol/ml), and 3.5 µl Lipofectamine RNAiMAX (Invitrogen, USA) and 250 µl Opti-MEM medium were mixed and reacted for 5 minutes at room temperature, followed by reaction for 20 minutes at room temperature. Subsequently, 500 µl of each of the transformation solutions was dispensed into the cultured tumor cell lines, and cultured for 5 hours, and then the medium mixed with the transformation solution was removed. Thereafter, 2.0 ml of the culture medium was dispensed into each well of each tumor cell line, and then cultured for 48 to 72 hours _{under conditions of 37° C. and 5% (v/v) CO 2} to obtain transformed cells. In the case of using plates of different sizes, transformation was performed by adjusting the number of cells and the amount of solution in proportion to the area of the well.

Example 3: Used to screen cancer cell growth inhibition miRNA Library selection

A total of 300 miRNAs were included in the miRNA library, and among them, miRNAs whose expression levels were changed in cancer tissues compared to normal tissues were first selected. In several cancers, there were 52 miRNAs that were commonly changed (29 miRNAs were reduced in expression compared to normal cells in cancer and 23 miRNAs were elevated), and 61 miRNAs were specifically changed in each cancer (breast cancer). ; 7 miRNAs, colon cancer; 5 miRNAs, liver cancer; 29 miRNAs, lung cancer; 8 miRNAs, prostate cancer; 12 miRNAs) (Int J Cancer, 2011, 129: 1624-1634, Human tumor microRNA signatures derived from large-scale oligonucleotide microarray datasets). The rest include genes related to cell cycle and cell death (CDK2, CDK4, CDK6, CCND1, CCND2, CCND3, CCNE1, CCNE2, E2F1, E2F2, E2F3, Plk1, Bcl-2, Bcl-xL, Mcl-1, BIRC2, BIRC3, BIRC5, XIAP) were selected and included miRNAs with high potential to target these genes with the TargetScan program. Table 1 shows the guide and passenger sequences of 300 miRNA mimics used for screening.

Example 4: inhibit cancer cell growth miRNA Screening

After transforming 300 miRNA mimics into A549 lung cancer cells at a concentration of 20 nM by the method of Example 2, after 72 hours, CCK-8 (Dojindo Laboratories, Japan) reagent was added to 1/10 of the medium and 1.5 in a 37°C incubator. After the time reaction, the absorbance was measured at 450 nm to compare the change in the number of cells. The results are shown as a graph in FIG. 1 and CCK-8 values in Table 2 are shown for each miRNA. As a control, the CCK-8 value of cancer cells transformed with the NC mimic (negative control mimic) was used as 100%, and the CCK-8 value of the cancer cells transformed with each miRNA mimic was shown relatively. When looking at miRNA mimics with good cancer cell growth inhibition, 49 were less than 70% and 5 were less than 60% compared to the control group. In particular, miR-608 showed better cancer cell growth inhibitory effect than miR-16 and miR-542-3p, which were used as positive controls. Selecting 5 miRNAs (miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608) with good cancer growth inhibitory effect (miR-28-5p, miR-323-5p, miR-608) to control cancer cell growth Follow-up studies such as identification were conducted.

NC mimic:

Sense RNA strand 5'-UUCUCCGAACGUGUCACGUTT-3' (SEQ ID NO: 601)

Antisense RNA strand 5'-ACGUGACACGUUCGGAGAATT-3' (SEQ ID NO: 602)

has-miR-16-5p:

guide RNA strand 5'-UAGCAGCACGUAAAUAUUGGCG-3' (SEQ ID NO: 603)

passenger RNA strand 5'-CCAAUAUUUACGUGCUGCUAUU-3' (SEQ ID NO: 604)

has-miR-542-3p

guide RNA strand 5'-UGUGACAGAUUGAUAACUGAAA-3' (MIMAT0003389, SEQ ID NO: 605)

passenger RNA strand 5'-UCAGUUAUCAAUCUGUCACAUU-3' (SEQ ID NO: 606)

Example 5: In cancer cell lines miR -28-5p, miR -323-5p, miR -510-5p, miR-552-3p, miR-608 On mimics Cell proliferation inhibitory effect

To verify the inhibitory effect of miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, and miR-608 mimics in cancer cell lines other than the A549 lung cancer cell line. Uterine cancer cells HeLa transformed by the method of Example 2, breast cancer cells MCF7, prostate cancer cells PC3, HeLa contaminated cells KB, lung cancer cells NCI-H460, liver cancer cells Hep3B, colon cancer cells HCT116 and normal cells, fibroblasts WI-38. After culturing for 3 to 5 days, the number of cells was counted using a hemocytometer, and then the change in the number of cells in each cancer cell line was compared (see FIG. 2). At this time, as a control, cancer cells transformed with an NC mimic (negative control mimic) were used.

2 is a human uterine cancer cell HeLa transformed with the double-stranded miRNAs of the present invention, breast cancer cell MCF7, prostate cancer cell PC3, HeLa contaminated cell KB, lung cancer cell NCI-H460, liver cancer cell Hep3B, colon cancer cell HCT116 and normal cells. It is a graph showing the change in the number of cells of the phosphorus fibroblast WI-38. As can be seen in Figure 2, there was a difference for each cancer cell, but as a whole, miR-323-5p and miR-608 showed good cell growth inhibitory effect. In NCI-H460, a lung cancer cell, miR-608 and miR-552-3p had a good inhibitory effect, and A549, another lung cancer cell, had a good inhibitory effect, so miR-552-3p and miR-608 are highly likely to be used for lung cancer treatment. do. In addition, miR-552-3p and miR-608 showed good growth inhibitory effects on liver cancer (Hep3B) and colon cancer (HCT116) cells, so it is thought that they can be used for the treatment of liver and colon cancer. In addition, miR-608 was shown to have a good growth inhibitory effect on prostate cancer PC3 and breast cancer MCF7 cells. miR-323-5p has a good inhibitory effect on uterine cancer cells HeLa, breast cancer cells MCF7 and prostate cancer cells PC3, so it can be used to inhibit the growth of these cells. WI-38 cells were normal cells, but miR-608 inhibited the growth of WI-38 cells, and miR-323-5p did not. Therefore, miR-323-5p is highly likely to have no effect on normal cells and is considered to be highly specific for cancer cells.

Example 6: Lung cancer cell line In A549 miR -28-5p, miR -323-5p, miR -510-5p, miR-552-3p, miR -608 On the mimic by Colony Formation ability Inhibitory effect

To verify the effect of inhibiting colony formation by the miRNA mimics of the present invention, lung cancer cells A549 transformed by the method of Example 2 were cultured for 24 hours, and then 200 cells were dispensed into a 6-well plate. , 37° C., 5% (v/v) CO ₂ was cultured for 10 days, stained with crystal violet, and then the number of colonies was counted and compared (see FIGS. 3A and 3B ). At this time, as a control, cancer cells transformed with an NC mimic (negative control mimic) were used. Figure 3a is a graph showing the number of colonies of lung cancer cell lines treated with miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 mimic and NC mimic, respectively, 3B is a photograph showing colonies of lung cancer cell lines treated with miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 mimic and NC mimic, respectively. As can be seen in Figures 3a and 3b, the experimental group treated with the miRNA mimics of the present invention (SEQ ID NOs: 33 to 34, SEQ ID NOs: 237 to 238, SEQ ID NOs: 365 to 366, SEQ ID NOs: 413 to 414 or SEQ ID NOs: 467 to 468) In the case of, it was found that the number of colonies was smaller than that of the cells treated with the control (NC mimic), and in particular, the colony-forming ability of lung cancer cells was effectively suppressed by miR-552-3p and miR-608.

Summarizing the results of Examples 4, 5, and 6, the double-stranded miRNAs of the present invention are various types of cancers, in particular miR-552-3p and miR-608 are lung cancer cells, liver cancer cells and colon cancer cells, and miR- Since 323-5p has an effect of inhibiting the proliferation of uterine cancer cells, breast cancer cells and prostate cancer cells, it can be seen that it can be used as a general anticancer agent.

Example 7: in lung cancer cell line A549 miR -28-5p, miR -323-5p, miR -510-5p, miR-552-3p, miR -608 On the mimic Cell cycle inhibition effect analysis

Targeting the A549 cells transformed by the method of Example 2, the effect of miRNA mimics on the cell cycle was analyzed by flow cytometry (FACS Vantage SE, Becton Dickinson). 48 and 72 hours after transformation, the cells were trypsinized to separate them into single cells, washed twice with PBS, stained with propidium iodide, and analyzed by flow cytometry (see Fig. 4, Table 3). At this time, the percentage for each stage of the cell cycle was measured using cell cycle analysis software (Cell Quest, Becton Dickinson). The number of nuclei is calculated by measuring the fluorescence stained on the nucleus in the cell using a flow cytometer. If the number of nuclei is 2N, it is classified as G0/G1 step, and if 4N, it is classified as S step. In addition, sub-G1 whose number of nuclei is less than 2N is classified as a cell death stage in which the cells die.

Figure 4 is a cell cycle measured by flow cytometry for lung cancer cell lines treated with miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 mimic and NC mimic, respectively. It is a step and the values of the number of cells in each step are shown in Table 3.

As can be seen in FIG. 4, it can be seen that the number of G1 cells increases when miR-28-5p, miR-323-5p, and miR-510-5p mimics are treated, and thus, cell cycle progression is stopped at the G1 stage, and thus the cells It was confirmed that growth was inhibited. When apoptosis occurs, the number of cells belonging to sub-G1 increases, and as shown in FIG. 4, apoptosis occurred in the order of miR-608> miR-552-3p> miR-323-5p> miR-28-5p. Able to know. In particular, in the case of miR-608, more than 30% of cells belong to sub-G1 even on the second day, and more than 60% of the cells belong to sub-G1 on the third day, so a large number of cells die when treated with miR-608 mimics. I could confirm.

Therefore, as can be seen in Figure 4 and Table 3, it can be seen that treatment with miR-28-5p, miR-323-5p, and miR-510-5p mimics inhibits cell proliferation by stopping cell growth in the G1 stage. It was found that more than half of the cells were killed when the miR-608 mimic was treated.

Example 8: Lung cancer cell line In A549 miR -28-5p, miR -323-5p, miR -510-5p, miR-552-3p, miR -608 On the mimic Apoptosis induction experiment by Annexin V staining)

Cell death by miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 mimics targeting A549 cells transformed by the method of Example 2 above. Annexin V staining (Annexin-V-FLUOS, Roche Applied Science, Germany) was performed to determine the induction.

Cells undergoing apoptosis undergo various changes, and one of them is the phenomenon that phosphatidylserine in the cell membrane is inside the membrane and then flipped outward. Since Annexin V binds to phosphatidylserine, the amount of phosphatidylserine released outside the cell can be determined by the degree of binding of fluorescently labeled Annexin V. 48 and 72 hours after transformation of lung cancer cells A549, Annexin-V-FLUOS labeling solution was added at a time of 100 µl/well (based on 24-well plate), reacted at room temperature for 15 minutes, and observed under a microscope. I did. First, looking at Figures 5a and 5b, it can be seen that the number of cells on the 2nd and 3rd day decreases when compared to NC. Especially on the 3rd day, when the anticancer drug camptothecin was treated and the miR-608 mimic was transformed It was found that few cells remained. Next, the number of cells of miR-552-3p and miR-323-5p was significantly reduced. On the second day, the Annexin V photo shows that many cells were combined with Annexin V in the case of camptothecin treatment, miR-552-3p, and miR-608, followed by Annexin V in the case of miR-323-5p. There were many cells. On the third day, in the case of camptothecin treatment and in the case of miR-608, the cells were almost completely dead and there were almost no remaining cells, so the fluorescently labeled cells were rather reduced. It has shrunk to some extent. When looking at the results of this Annexin V assay, it was found that miR-323-5p, miR-552-3p, and miR-608 induce apoptosis in A549 cells.

Example 9: Western Blot ( western blot ) Through analysis target excavation

Western blotting was performed on A549 cells transformed by the method of Example 2, using antibodies against proteins related to cell death and cell cycle. Specifically, the entire intracellular protein was extracted from A549 cells 48 hours after transformation using a protein extraction kit (PRO-PREP protein extraction reagent, Intron Biotechnology, Korea). SDS sample buffer solution (Komabiotech, Korea) was added to 30 µg of the extracted total protein and treated at 95° C. for 5 minutes, followed by 4-12% SDS polyacrylamide gel electrophoresis (SDS-PAGE). Then, the separated proteins were transferred to a nitrocellulose membrane by electrophoresis, and then reacted for 1 hour in a Tris buffer solution containing 5% skim milk (Tris buffered saline + 0.05% Tween 20). Thereafter, monoclonal antibodies against proteins related to cell death and cell cycle (Cell Signaling Technology, Danvers, MA; Santa Cruz Biotechnology, Dallas, Texas) were used as primary antibodies, and horseradish peroxidase was used. Western blot was performed by sequentially binding the bound anti-rabbit or anti-mouse antibody (Cell Signaling Technology, Danvers, MA) as a secondary antibody and then inducing a luminescence reaction with an ECL detection kit (Amersham Biosciences, UK). (See: Figures 6a, 6b and 6c).

Figure 6a is a Western blot photograph showing the levels of proteins related to apoptosis expressed in cells transformed with the double-stranded miRNA of the present invention, Figure 6b is a cell cycle expressed in cells transformed with the double-stranded miRNA of the present invention It is a Western blot photograph showing the protein levels related to and FIG. 6C is a Western blot photograph showing the protein levels related to the Akt pathway expressed in cells transformed with the double-stranded miRNA of the present invention. As can be seen in Figure 6a, the expression of anti-apoptotic proteins was inhibited by the miRNAs of the present invention, specifically miR-323-5p is Bcl-xL and BIRC5, miR-510-5p is Mcl-1 and BIRC5, miR-552-3p decreased the protein expression levels of Mcl-1 and BIRC5, and miR-608 decreased the protein expression level of Bcl-xL. miR-28-5p, miR-510-5p, and miR-608 were the apoptosis-inducing protein BIK. The amount of protein expression was increased. In addition, as shown in Figure 6b, the expression of proteins required for cell cycle progression was suppressed by the miRNAs of the present invention, specifically miR-28-5p is CCND3, miR-323-5p is CDK2, CDK4, CCND1, E2F1, miR -552-3p decreased the protein expression levels of CDK6 and CCND3, and miR-608 decreased the protein expression levels of CDK6, CCND1, and CCND3. In Figure 6c, the activation of the Akt pathway required for cell survival was investigated, and it was found that the phosphorylation of Ser473 and Thr308, which activates Akt, was significantly reduced when the miRNAs of the present invention were transformed, and the regulation of PI3K, an enzyme required for the activation of Akt It was confirmed that the expression level of the subunit PIK3R2 protein was significantly reduced by miR-323-5p and miR-608. In addition, Sp1, a transcription factor that widely affects cell differentiation, growth, and death, was also inhibited by miR-28-5p, miR-323-5p, miR-510-5p, and miR-608. Therefore, as can be seen in Figures 6a, 6b and 6c, miR-28-5p, miR-323-5p, miR-510-5p, miR-552-3p, miR-608 mimetics (SEQ ID NOs: 33 to 34, sequence Numbers 237 to 238, SEQ ID NOs: 365 to 366, SEQ ID NOs: 413 to 414 or SEQ ID NOs: 467 to 468) were confirmed to simultaneously inhibit one or more proteins required for cell survival and growth, and this is shown in Table 4. Indicated.

Example 10: target For genes siRNA Cell growth inhibition experiment of lung cancer cell A549 using

72 hours after the siRNAs were transformed into lung cancer cells A549 by the method of Example 2, the number of cells was measured using a hemocytometer. The siRNAs used were siRNAs that inhibit the expression of CCND1, CCND3, CDK6, CYB5R3, Bcl-xL, Mcl-1, BIRC5, PIK3R2, and Sp1 genes. The base sequences are shown in Table 5.

As can be seen in Figure 7, it can be seen that cell growth is inhibited when siRNAs that suppress the expression of Bcl-xL, BIRC5, CCND3, CYB5R3, Mcl-1, and PIK3R2 genes are inhibited. It was confirmed that the suppressing miRNAs of the present invention can effectively inhibit cancer cell growth.

Example 11: miR -608 Mimic Transformed human lung cancer cells NCI - H460 Inhibitory Effect of Tumor Proliferation in Nude Mouse

The experimental animals were 10 female nude mice (Nudes-nu/nu mice, BALB/c/Bkl background, female) with an average weight of around 19.3 ± 1 g at 6 weeks of age (Nara Biotech, Korea) for a week of adaptation. It was used after roughing. Nude mice were bred in the SPF animal laboratory, and the temperature was 21.0 ~ 23.0℃ and the humidity was maintained at 45.0%.

Using the miR-608 mimic (SEQ ID NO: 467 to 468) prepared in Example 1, human lung cancer cells transformed with miR-608 mimic (SEQ ID NO: 467 to 468) or control NC double-stranded RNA (NCI -H460) was cultured for 24 hours, then xenograft was performed by subcutaneous injection of ^{5 X 10 6 cells into the neck of the nude mouse, 5 heads for each sample.} Tumor size was measured on the 5th, 8th, 12th, 15th, 19th, 22nd, and 26th after transplantation, and the long and short diameters were measured and'average tumor size = [(short diameter) x (short diameter)] It was calculated as x (longer diameter)/2 (mm ³ )', and the tumor sizes of each experimental group and control group were averaged and displayed as a graph, and the difference in tumor size between each group was confirmed (see Fig. 8). At this time, the statistical significance of the change in tumor size between the tumor cell inoculation group transformed with the miR-608 mimic or the tumor cell inoculation group transformed with the control NC double-stranded RNA was analyzed by Student's t-test, and the probability of significance (P When value) was less than 0.05, it was recognized as having statistical significance.

8 is a graph showing the change in tumor size over time in nude mice transplanted with tumor cells transformed with double-stranded miRNA of the present invention, (■) is a tumor transformed by miR-608 mimic It represents a nude mouse transplanted with cells, and (▲) represents a control group.

As can be seen in FIG. 8, a large difference could not be confirmed in each experimental group until 8 days after transplantation, but after 8 days, the tumor size of the tumor cell inoculation group transformed with the miR-608 mimic was compared with the tumor size of the control group. Remarkably small ones could be observed, and thus, when transformed with miR-608 mimics, remarkable inhibitory activity of tumor growth could be confirmed.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in other specific forms without changing the technical spirit or essential features thereof. In this regard, the embodiments described above are illustrative in all respects and should be understood as non-limiting. The scope of the present invention should be construed that all changes or modifications derived from the meaning and scope of the claims to be described later rather than the above detailed description and equivalent concepts are included in the scope of the present invention.

<110> University-Industry Cooperation Group of Kyung Hee University <120> Pharmaceutical composition for preventing and treating cancer comprising double-stranded miRNAs as active ingredient <130> PA131399-KR-P1-D1 <150> KR10-2014-0016782 <151> 2014-02-13 <160> 624 <170> KopatentIn 2.0 <210> 1 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-7-5p guide sequence <400> 1 uggaagacua gugauuuugu ugu 23 <210> 2 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-7-5p passenger sequence <400> 2 aacaaaauca cuagucuucc auu 23 <210> 3 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-9-5p guide sequence <400> 3 ucuuugguua ucuagcugua uga 23 <210> 4 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-9-5p passenger sequence <400> 4 auacagcuag auaaccaaag auu 23 <210> 5 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-9-3p guide sequence <400> 5 auaaagcuag auaaccgaaa gu 22 <210> 6 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-9-3p passenger sequence <400> 6 uuucgguuau cuagcuuuau uu 22 <210> 7 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-10b-5p guide sequence <400> 7 uacccuguag aaccgaauuu gug 23 <210> 8 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-10b-5p passenger sequence <400> 8 caaauucggu ucuacagggu auu 23 <210> 9 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-10b-3p guide sequence <400> 9 acagauucga uucuagggga au 22 <210> 10 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-10b-3p passenger sequence <400> 10 uccccuagaa ucgaaucugu uu 22 <210> 11 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-17-3p guide sequence <400> 11 acugcaguga aggcacuugu ag 22 <210> 12 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-17-3p passenger sequence <400> 12 acaagugccu ucacugcagu uu 22 <210> 13 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-19a-3p guide sequence <400> 13 ugugcaaauc uaugcaaaac uga 23 <210> 14 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-19a-3p passenger sequence <400> 14 aguuuugcau agauuugcac auu 23 <210> 15 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-20a-5p guide sequence <400> 15 uaaagugcuu auagugcagg uag 23 <210> 16 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-20a-5p passenger sequence <400> 16 accugcacua uaagcacuuu auu 23 <210> 17 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-21-5p guide sequence <400> 17 uagcuuauca gacugauguu ga 22 <210> 18 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-21-5p passenger sequence <400> 18 aacaucaguc ugauaagcua uu 22 <210> 19 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-22-3p guide sequence <400> 19 aagcugccag uugaagaacu gu 22 <210> 20 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-22-3p passenger sequence <400> 20 aguucuucaa cuggcagcuu uu 22 <210> 21 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-23a-3p guide sequence <400> 21 aucacauugc cagggauuuc c 21 <210> 22 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-23a-3p passenger sequence <400> 22 aaaucccugg caaugugauu u 21 <210> 23 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-24-3p guide sequence <400> 23 uggcucaguu cagcaggaac ag 22 <210> 24 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-24-3p passenger sequence <400> 24 guuccugcug aacugagcca uu 22 <210> 25 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-24-1-5p guide sequence <400> 25 ugccuacuga gcugauauca gu 22 <210> 26 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-24-1-5p passenger sequence <400> 26 ugauaucagc ucaguaggca uu 22 <210> 27 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-25-3p guide sequence <400> 27 cauugcacuu gucucggucu ga 22 <210> 28 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-25-3p passenger sequence <400> 28 agaccgagac aagugcaaug uu 22 <210> 29 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-27a-3p guide sequence <400> 29 uucacagugg cuaaguuccg c 21 <210> 30 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-27a-3p passenger sequence <400> 30 ggaacuuagc cacugugaau u 21 <210> 31 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-27b-3p guide sequence <400> 31 uucacagugg cuaaguucug c 21 <210> 32 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-27b-3p passenger sequence <400> 32 agaacuuagc cacugugaau u 21 <210> 33 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-28-5p guide sequence <400> 33 aaggagcuca cagucuauug ag 22 <210> 34 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-28-5p passenger sequence <400> 34 caauagacug ugagcuccuu uu 22 <210> 35 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-30a-5p guide sequence <400> 35 uguaaacauc cucgacugga ag 22 <210> 36 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-30a-5p passenger sequence <400> 36 uccagucgag gauguuuaca uu 22 <210> 37 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-30a-3p guide sequence <400> 37 cuuucagucg gauguuugca gc 22 <210> 38 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-30a-3p passenger sequence <400> 38 ugcaaacauc cgacugaaag uu 22 <210> 39 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-30b-5p guide sequence <400> 39 uguaaacauc cuacacucag cu 22 <210> 40 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-30b-5p passenger sequence <400> 40 cugaguguag gauguuuaca uu 22 <210> 41 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-30c-1-3p guide sequence <400> 41 cugggagagg guuguuuacu cc 22 <210> 42 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-30c-1-3p passenger sequence <400> 42 aguaaacaac ccucucccag uu 22 <210> 43 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-30e-3p guide sequence <400> 43 cuuucagucg gauguuuaca gc 22 <210> 44 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-30e-3p passenger sequence <400> 44 uguaaacauc cgacugaaag uu 22 <210> 45 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-31-5p guide sequence <400> 45 aggcaagaug cuggcauagc u 21 <210> 46 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-31-5p passenger sequence <400> 46 cuaugccagc aucuugccuu u 21 <210> 47 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-32-5p guide sequence <400> 47 uauugcacau uacuaaguug ca 22 <210> 48 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-32-5p passenger sequence <400> 48 caacuuagua augugcaaua uu 22 <210> 49 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-33a-5p guide sequence <400> 49 gugcauugua guugcauugc a 21 <210> 50 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-33a-5p passenger sequence <400> 50 caaugcaacu acaaugcacu u 21 <210> 51 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-34a-5p guide sequence <400> 51 uggcaguguc uuagcugguu gu 22 <210> 52 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-34a-5p passenger sequence <400> 52 aaccagcuaa gacacugcca uu 22 <210> 53 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-34b-3p guide sequence <400> 53 caaucacuaa cuccacugcc au 22 <210> 54 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-34b-3p passenger sequence <400> 54 ggcaguggag uuagugauug uu 22 <210> 55 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-34b-5p guide sequence <400> 55 uaggcagugu cauuagcuga uug 23 <210> 56 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-34b-5p passenger sequence <400> 56 aucagcuaau gacacugccu auu 23 <210> 57 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-34c-5p guide sequence <400> 57 aggcagugua guuagcugau ugc 23 <210> 58 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-34c-5p passenger sequence <400> 58 aaucagcuaa cuacacugcc uuu 23 <210> 59 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-34c-3p guide sequence <400> 59 aaucacuaac cacacggcca gg 22 <210> 60 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-34c-3p passenger sequence <400> 60 uggccgugug guuagugauu uu 22 <210> 61 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-92a-3p guide sequence <400> 61 uauugcacuu gucccggccu gu 22 <210> 62 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-92a-3p passenger sequence <400> 62 aggccgggac aagugcaaua uu 22 <210> 63 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-93-5p guide sequence <400> 63 caaagugcug uucgugcagg uag 23 <210> 64 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-93-5p passenger sequence <400> 64 accugcacga acagcacuuu guu 23 <210> 65 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-95-3p guide sequence <400> 65 uucaacgggu auuuauugag ca 22 <210> 66 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-95-3p passenger sequence <400> 66 cucaauaaau acccguugaa uu 22 <210> 67 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-96-5p guide sequence <400> 67 uuuggcacua gcacauuuuu gcu 23 <210> 68 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-96-5p passenger sequence <400> 68 caaaaaugug cuagugccaa auu 23 <210> 69 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-99a-5p guide sequence <400> 69 aacccguaga uccgaucuug ug 22 <210> 70 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-99a-5p passenger sequence <400> 70 caagaucgga ucuacggguu uu 22 <210> 71 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-99b-5p guide sequence <400> 71 cacccguaga accgaccuug cg 22 <210> 72 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-99b-5p passenger sequence <400> 72 caaggucggu ucuacgggug uu 22 <210> 73 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-100-5p guide sequence <400> 73 aacccguaga uccgaacuug ug 22 <210> 74 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-100-5p passenger sequence <400> 74 caaguucgga ucuacggguu uu 22 <210> 75 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-101-3p guide sequence <400> 75 uacaguacug ugauaacuga a 21 <210> 76 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-101-3p passenger sequence <400> 76 caguuaucac aguacuguau u 21 <210> 77 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-105-5p guide sequence <400> 77 ucaaaugcuc agacuccugu ggu 23 <210> 78 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-105-5p passenger sequence <400> 78 cacaggaguc ugagcauuug auu 23 <210> 79 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-106b-5p guide sequence <400> 79 uaaagugcug acagugcaga u 21 <210> 80 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-106b-5p passenger sequence <400> 80 cugcacuguc agcacuuuau u 21 <210> 81 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-107 guide sequence <400> 81 agcagcauug uacagggcua uca 23 <210> 82 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-107 passenger sequence <400> 82 auagcccugu acaaugcugc uuu 23 <210> 83 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-122-5p guide sequence <400> 83 uggaguguga caaugguguu ug 22 <210> 84 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-122-5p passenger sequence <400> 84 aacaccauug ucacacucca uu 22 <210> 85 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-125a-5p guide sequence <400> 85 ucccugagac ccuuuaaccu guga 24 <210> 86 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-125a-5p passenger sequence <400> 86 acagguuaaa gggucucagg gauu 24 <210> 87 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-125a-3p guide sequence <400> 87 acaggugagg uucuugggag cc 22 <210> 88 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-125a-3p passenger sequence <400> 88 cucccaagaa ccucaccugu uu 22 <210> 89 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-125b-5p guide sequence <400> 89 ucccugagac ccuaacuugu ga 22 <210> 90 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-125b-5p passenger sequence <400> 90 acaaguuagg gucucaggga uu 22 <210> 91 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-126-3p guide sequence <400> 91 ucguaccgug aguaauaaug cg 22 <210> 92 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-126-3p passenger sequence <400> 92 cauuauuacu cacgguacga uu 22 <210> 93 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-126-5p guide sequence <400> 93 cauuauuacu uuugguacgc g 21 <210> 94 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-126-5p passenger sequence <400> 94 cguaccaaaa guaauaaugu u 21 <210> 95 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-127-5p guide sequence <400> 95 cugaagcuca gagggcucug au 22 <210> 96 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-127-5p passenger sequence <400> 96 cagagcccuc ugagcuucag uu 22 <210> 97 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-128-3p guide sequence <400> 97 ucacagugaa ccggucucuu u 21 <210> 98 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-128-3p passenger sequence <400> 98 agagaccggu ucacugugau u 21 <210> 99 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-129-5p guide sequence <400> 99 cuuuuugcgg ucugggcuug c 21 <210> 100 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-129-5p passenger sequence <400> 100 aagcccagac cgcaaaaagu u 21 <210> 101 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-130a-3p guide sequence <400> 101 cagugcaaug uuaaaagggc au 22 <210> 102 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-130a-3p passenger sequence <400> 102 gcccuuuuaa cauugcacug uu 22 <210> 103 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-132-3p guide sequence <400> 103 uaacagucua cagccauggu cg 22 <210> 104 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-132-3p passenger sequence <400> 104 accauggcug uagacuguua uu 22 <210> 105 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-132-5p guide sequence <400> 105 accguggcuu ucgauuguua cu 22 <210> 106 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-132-5p passenger sequence <400> 106 uaacaaucga aagccacggu uu 22 <210> 107 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-133a-3p guide sequence <400> 107 uuuggucccc uucaaccagc ug 22 <210> 108 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-133a-3p passenger sequence <400> 108 gcugguugaa ggggaccaaa uu 22 <210> 109 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-134-5p guide sequence <400> 109 ugugacuggu ugaccagagg gg 22 <210> 110 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-134-5p passenger sequence <400> 110 ccucugguca accagucaca uu 22 <210> 111 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-135a-5p guide sequence <400> 111 uauggcuuuu uauuccuaug uga 23 <210> 112 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-135a-5p passenger sequence <400> 112 acauaggaau aaaaagccau auu 23 <210> 113 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-136-5p guide sequence <400> 113 acuccauuug uuuugaugau gga 23 <210> 114 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-136-5p passenger sequence <400> 114 caucaucaaa acaaauggag uuu 23 <210> 115 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-137 guide sequence <400> 115 uuauugcuua agaauacgcg uag 23 <210> 116 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-137 passenger sequence <400> 116 acgcguauuc uuaagcaaua auu 23 <210> 117 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-139-5p guide sequence <400> 117 ucuacagugc acgugucucc ag 22 <210> 118 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-139-5p passenger sequence <400> 118 ggagacacgu gcacuguaga uu 22 <210> 119 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-140-5p guide sequence <400> 119 cagugguuuu acccuauggu ag 22 <210> 120 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-140-5p passenger sequence <400> 120 accauagggu aaaaccacug uu 22 <210> 121 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-140-3p guide sequence <400> 121 uaccacaggg uagaaccacg g 21 <210> 122 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-140-3p passenger sequence <400> 122 gugguucuac ccugugguau u 21 <210> 123 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-142-5p guide sequence <400> 123 cauaaaguag aaagcacuac u 21 <210> 124 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-142-5p passenger sequence <400> 124 uagugcuuuc uacuuuaugu u 21 <210> 125 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-142-3p guide sequence <400> 125 uguaguguuu ccuacuuuau gga 23 <210> 126 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-142-3p passenger sequence <400> 126 cauaaaguag gaaacacuac auu 23 <210> 127 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-143-3p guide sequence <400> 127 ugagaugaag cacuguagcu c 21 <210> 128 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-143-3p passenger sequence <400> 128 gcuacagugc uucaucucau u 21 <210> 129 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-144-3p guide sequence <400> 129 uacaguauag augauguacu 20 <210> 130 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-144-3p passenger sequence <400> 130 uacaucaucu auacuguauu 20 <210> 131 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-145-5p guide sequence <400> 131 guccaguuuu cccaggaauc ccu 23 <210> 132 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-145-5p passenger sequence <400> 132 ggauuccugg gaaaacugga cuu 23 <210> 133 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-146a-5p guide sequence <400> 133 ugagaacuga auuccauggg uu 22 <210> 134 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-146a-5p passenger sequence <400> 134 cccauggaau ucaguucuca uu 22 <210> 135 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-147a guide sequence <400> 135 guguguggaa augcuucugc 20 <210> 136 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-147a passenger sequence <400> 136 agaagcauuu ccacacacuu 20 <210> 137 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-149-5p guide sequence <400> 137 ucuggcuccg ugucuucacu ccc 23 <210> 138 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-149-5p passenger sequence <400> 138 gagugaagac acggagccag auu 23 <210> 139 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-150-5p guide sequence <400> 139 ucucccaacc cuuguaccag ug 22 <210> 140 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-150-5p passenger sequence <400> 140 cugguacaag gguugggaga uu 22 <210> 141 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-151-5p guide sequence <400> 141 ucgaggagcu cacagucuag u 21 <210> 142 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-151-5p passenger sequence <400> 142 uagacuguga gcuccucgau u 21 <210> 143 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-152-3p guide sequence <400> 143 ucagugcaug acagaacuug g 21 <210> 144 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-152-3p passenger sequence <400> 144 aaguucuguc augcacugau u 21 <210> 145 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-153-3p guide sequence <400> 145 uugcauaguc acaaaaguga uc 22 <210> 146 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-153-3p passenger sequence <400> 146 ucacuuuugu gacuaugcaa uu 22 <210> 147 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-154-5p guide sequence <400> 147 uagguuaucc guguugccuu cg 22 <210> 148 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-154-5p passenger sequence <400> 148 aaggcaacac ggauaaccua uu 22 <210> 149 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-154-3p guide sequence <400> 149 aaucauacac gguugaccua uu 22 <210> 150 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-154-3p passenger sequence <400> 150 uaggucaacc guguaugauu uu 22 <210> 151 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-155-5p guide sequence <400> 151 uuaaugcuaa ucgugauagg ggu 23 <210> 152 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-155-5p passenger sequence <400> 152 cccuaucacg auuagcauua auu 23 <210> 153 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-181a-5p guide sequence <400> 153 aacauucaac gcugucggug agu 23 <210> 154 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-181a-5p passenger sequence <400> 154 ucaccgacag cguugaaugu uuu 23 <210> 155 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-183-5p guide sequence <400> 155 uauggcacug guagaauuca cu 22 <210> 156 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-183-5p passenger sequence <400> 156 ugaauucuac cagugccaua uu 22 <210> 157 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-183-3p guide sequence <400> 157 gugaauuacc gaagggccau aa 22 <210> 158 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-183-3p passenger sequence <400> 158 auggcccuuc gguaauucac uu 22 <210> 159 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-184 guide sequence <400> 159 uggacggaga acugauaagg gu 22 <210> 160 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-184 passenger sequence <400> 160 ccuuaucagu ucuccgucca uu 22 <210> 161 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-185-5p guide sequence <400> 161 uggagagaaa ggcaguuccu ga 22 <210> 162 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-185-5p passenger sequence <400> 162 aggaacugcc uuucucucca uu 22 <210> 163 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-187-3p guide sequence <400> 163 ucgugucuug uguugcagcc gg 22 <210> 164 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-187-3p passenger sequence <400> 164 ggcugcaaca caagacacga uu 22 <210> 165 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-187-3p passenger sequence <400> 165 caacggaauc ccaaaagcag cug 23 <210> 166 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-187-3p passenger sequence <400> 166 gcugcuuuug ggauuccguu guu 23 <210> 167 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-192-5p guide sequence <400> 167 cugaccuaug aauugacagc c 21 <210> 168 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-192-5p passenger sequence <400> 168 cugucaauuc auaggucagu u 21 <210> 169 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-193a-5p guide sequence <400> 169 ugggucuuug cgggcgagau ga 22 <210> 170 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-193a-5p passenger sequence <400> 170 aucucgcccg caaagaccca uu 22 <210> 171 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-193a-3p guide sequence <400> 171 aacuggccua caaaguccca gu 22 <210> 172 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-193a-3p passenger sequence <400> 172 ugggacuuug uaggccaguu uu 22 <210> 173 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-193b-3p guide sequence <400> 173 aacuggcccu caaagucccg cu 22 <210> 174 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-193b-3p passenger sequence <400> 174 cgggacuuug agggccaguu uu 22 <210> 175 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-194-5p guide sequence <400> 175 uguaacagca acuccaugug ga 22 <210> 176 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-194-5p passenger sequence <400> 176 cacauggagu ugcuguuaca uu 22 <210> 177 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-196a-5p guide sequence <400> 177 uagguaguuu cauguuguug gg 22 <210> 178 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-196a-5p passenger sequence <400> 178 caacaacaug aaacuaccua uu 22 <210> 179 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-197-3p guide sequence <400> 179 uucaccaccu ucuccaccca gc 22 <210> 180 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-197-3p passenger sequence <400> 180 uggguggaga agguggugaa uu 22 <210> 181 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-198 guide sequence <400> 181 gguccagagg ggagauaggu uc 22 <210> 182 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-198 passenger sequence <400> 182 accuaucucc ccucuggacc uu 22 <210> 183 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-199b-5p guide sequence <400> 183 cccaguguuu agacuaucug uuc 23 <210> 184 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-199b-5p passenger sequence <400> 184 acagauaguc uaaacacugg guu 23 <210> 185 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-202-3p guide sequence <400> 185 agagguauag ggcaugggaa 20 <210> 186 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-202-3p passenger sequence <400> 186 cccaugcccu auaccucuuu 20 <210> 187 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-202-5p guide sequence <400> 187 uuccuaugca uauacuucuu ug 22 <210> 188 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-202-5p passenger sequence <400> 188 aagaaguaua ugcauaggaa uu 22 <210> 189 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-204-5p guide sequence <400> 189 uucccuuugu cauccuaugc cu 22 <210> 190 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-204-5p passenger sequence <400> 190 gcauaggaug acaaagggaa uu 22 <210> 191 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-205-5p guide sequence <400> 191 uccuucauuc caccggaguc ug 22 <210> 192 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-205-5p passenger sequence <400> 192 gacuccggug gaaugaagga uu 22 <210> 193 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-206 guide sequence <400> 193 uggaauguaa ggaagugugu gg 22 <210> 194 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-206 passenger sequence <400> 194 acacacuucc uuacauucca uu 22 <210> 195 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-210-3p guide sequence <400> 195 cugugcgugu gacagcggcu ga 22 <210> 196 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-210-3p passenger sequence <400> 196 agccgcuguc acacgcacag uu 22 <210> 197 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-212-3p guide sequence <400> 197 uaacagucuc cagucacggc c 21 <210> 198 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-212-3p passenger sequence <400> 198 ccgugacugg agacuguuau u 21 <210> 199 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-216a-5p guide sequence <400> 199 uaaucucagc uggcaacugu ga 22 <210> 200 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-216a-5p passenger sequence <400> 200 acaguugcca gcugagauua uu 22 <210> 201 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-216b-5p guide sequence <400> 201 aaaucucugc aggcaaaugu ga 22 <210> 202 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-216b-5p passenger sequence <400> 202 acauuugccu gcagagauuu uu 22 <210> 203 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-217 guide sequence <400> 203 uacugcauca ggaacugauu gga 23 <210> 204 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-217 passenger sequence <400> 204 caaucaguuc cugaugcagu auu 23 <210> 205 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-218-5p guide sequence <400> 205 uugugcuuga ucuaaccaug u 21 <210> 206 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-218-5p passenger sequence <400> 206 augguuagau caagcacaau u 21 <210> 207 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-219a-5p guide sequence <400> 207 ugauugucca aacgcaauuc u 21 <210> 208 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-219a-5p passenger sequence <400> 208 aauugcguuu ggacaaucau u 21 <210> 209 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-219a-1-3p guide sequence <400> 209 agaguugagu cuggacgucc cg 22 <210> 210 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-219a-1-3p passenger sequence <400> 210 ggacguccag acucaacucu uu 22 <210> 211 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-221-3p guide sequence <400> 211 agcuacauug ucugcugggu uuc 23 <210> 212 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-221-3p passenger sequence <400> 212 aacccagcag acaauguagc uuu 23 <210> 213 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-222-3p guide sequence <400> 213 agcuacaucu ggcuacuggg u 21 <210> 214 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-222-3p passenger sequence <400> 214 ccaguagcca gauguagcuu u 21 <210> 215 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-222-5p guide sequence <400> 215 cucaguagcc aguguagauc cu 22 <210> 216 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-222-5p passenger sequence <400> 216 gaucuacacu ggcuacugag uu 22 <210> 217 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-223-3p guide sequence <400> 217 ugucaguuug ucaaauaccc ca 22 <210> 218 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-223-3p passenger sequence <400> 218 ggguauuuga caaacugaca uu 22 <210> 219 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-224-5p guide sequence <400> 219 caagucacua gugguuccgu u 21 <210> 220 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-224-5p passenger sequence <400> 220 cggaaccacu agugacuugu u 21 <210> 221 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-296-5p guide sequence <400> 221 agggcccccc cucaauccug u 21 <210> 222 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-296-5p passenger sequence <400> 222 aggauugagg gggggcccuu u 21 <210> 223 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-297 guide sequence <400> 223 auguaugugu gcaugugcau g 21 <210> 224 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-297 passenger sequence <400> 224 ugcacaugca cacauacauu u 21 <210> 225 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-298 guide sequence <400> 225 agcagaagca gggagguucu ccca 24 <210> 226 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-298 passenger sequence <400> 226 ggagaaccuc ccugcuucug cuuu 24 <210> 227 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-299-5p guide sequence <400> 227 ugguuuaccg ucccacauac au 22 <210> 228 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-299-5p passenger sequence <400> 228 guauguggga cgguaaacca uu 22 <210> 229 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-301a-3p guide sequence <400> 229 cagugcaaua guauugucaa agc 23 <210> 230 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-301a-3p passenger sequence <400> 230 uuugacaaua cuauugcacu guu 23 <210> 231 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-302c-3p guide sequence <400> 231 uaagugcuuc cauguuucag ugg 23 <210> 232 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-302c-3p passenger sequence <400> 232 acugaaacau ggaagcacuu auu 23 <210> 233 <211> 17 <212> RNA <213> Artificial Sequence <220> <223> has-miR-302f guide sequence <400> 233 uaauugcuuc cauguuu 17 <210> 234 <211> 17 <212> RNA <213> Artificial Sequence <220> <223> has-miR-302f passenger sequence <400> 234 acauggaagc aauuauu 17 <210> 235 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-320a guide sequence <400> 235 aaaagcuggg uugagagggc ga 22 <210> 236 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-320a passenger sequence <400> 236 gcccucucaa cccagcuuuu uu 22 <210> 237 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-323a-5p guide sequence <400> 237 aggugguccg uggcgcguuc gc 22 <210> 238 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-323a-5p passenger sequence <400> 238 gaacgcgcca cggaccaccu uu 22 <210> 239 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-323a-3p guide sequence <400> 239 cacauuacac ggucgaccuc u 21 <210> 240 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-323a-3p passenger sequence <400> 240 aggucgaccg uguaaugugu u 21 <210> 241 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-324-3p guide sequence <400> 241 acugccccag gugcugcugg 20 <210> 242 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-324-3p passenger sequence <400> 242 agcagcaccu ggggcaguuu 20 <210> 243 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-326 guide sequence <400> 243 ccucugggcc cuuccuccag 20 <210> 244 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-326 passenger sequence <400> 244 ggaggaaggg cccagagguu 20 <210> 245 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-328-3p guide sequence <400> 245 cuggcccucu cugcccuucc gu 22 <210> 246 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-328-3p passenger sequence <400> 246 ggaagggcag agagggccag uu 22 <210> 247 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-329-3p guide sequence <400> 247 aacacaccug guuaaccucu uu 22 <210> 248 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-329-3p passenger sequence <400> 248 agagguuaac cagguguguu uu 22 <210> 249 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-330-5p guide sequence <400> 249 ucucugggcc ugugucuuag gc 22 <210> 250 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-330-5p passenger sequence <400> 250 cuaagacaca ggcccagaga uu 22 <210> 251 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-330-3p guide sequence <400> 251 gcaaagcaca cggccugcag aga 23 <210> 252 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-330-3p passenger sequence <400> 252 ucugcaggcc gugugcuuug cuu 23 <210> 253 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-331-5p guide sequence <400> 253 cuagguaugg ucccagggau cc 22 <210> 254 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-331-5p passenger sequence <400> 254 aucccuggga ccauaccuag uu 22 <210> 255 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-335-5p guide sequence <400> 255 ucaagagcaa uaacgaaaaa ugu 23 <210> 256 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-335-5p passenger sequence <400> 256 auuuuucguu auugcucuug auu 23 <210> 257 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-335-3p guide sequence <400> 257 uuuuucauua uugcuccuga cc 22 <210> 258 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-335-3p passenger sequence <400> 258 ucaggagcaa uaaugaaaaa uu 22 <210> 259 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-338-5p guide sequence <400> 259 aacaauaucc uggugcugag ug 22 <210> 260 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-338-5p passenger sequence <400> 260 cucagcacca ggauauuguu uu 22 <210> 261 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-340-5p guide sequence <400> 261 uuauaaagca augagacuga uu 22 <210> 262 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-340-5p passenger sequence <400> 262 ucagucucau ugcuuuauaa uu 22 <210> 263 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-342-5p guide sequence <400> 263 aggggugcua ucugugauug a 21 <210> 264 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-342-5p passenger sequence <400> 264 aaucacagau agcaccccuu u 21 <210> 265 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-342-3p guide sequence <400> 265 ucucacacag aaaucgcacc cgu 23 <210> 266 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-342-3p passenger sequence <400> 266 gggugcgauu ucugugugag auu 23 <210> 267 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-345-5p guide sequence <400> 267 gcugacuccu aguccagggc uc 22 <210> 268 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-345-5p passenger sequence <400> 268 gcccuggacu aggagucagc uu 22 <210> 269 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-346 guide sequence <400> 269 ugucugcccg caugccugcc ucu 23 <210> 270 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-346 passenger sequence <400> 270 aggcaggcau gcgggcagac auu 23 <210> 271 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-361-3p guide sequence <400> 271 ucccccaggu gugauucuga uuu 23 <210> 272 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-361-3p passenger sequence <400> 272 aucagaauca caccuggggg auu 23 <210> 273 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-362-5p guide sequence <400> 273 aauccuugga accuaggugu gagu 24 <210> 274 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-362-5p passenger sequence <400> 274 ucacaccuag guuccaagga uuuu 24 <210> 275 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-362-3p guide sequence <400> 275 aacacaccua uucaaggauu ca 22 <210> 276 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-362-3p passenger sequence <400> 276 aauccuugaa uagguguguu uu 22 <210> 277 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-363-3p guide sequence <400> 277 aauugcacgg uauccaucug ua 22 <210> 278 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-363-3p passenger sequence <400> 278 cagauggaua ccgugcaauu uu 22 <210> 279 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-367-3p guide sequence <400> 279 aauugcacuu uagcaauggu ga 22 <210> 280 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-367-3p passenger sequence <400> 280 accauugcua aagugcaauu uu 22 <210> 281 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-369-5p guide sequence <400> 281 agaucgaccg uguuauauuc gc 22 <210> 282 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-369-5p passenger sequence <400> 282 gaauauaaca cggucgaucu uu 22 <210> 283 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-369-3p guide sequence <400> 283 aauaauacau gguugaucuu u 21 <210> 284 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-369-3p passenger sequence <400> 284 agaucaacca uguauuauuu u 21 <210> 285 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-370-3p guide sequence <400> 285 gccugcuggg guggaaccug gu 22 <210> 286 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-370-3p passenger sequence <400> 286 cagguuccac cccagcaggc uu 22 <210> 287 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-373-5p guide sequence <400> 287 acucaaaaug ggggcgcuuu cc 22 <210> 288 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-373-5p passenger sequence <400> 288 aaagcgcccc cauuuugagu uu 22 <210> 289 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-374a-5p guide sequence <400> 289 uuauaauaca accugauaag ug 22 <210> 290 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-374a-5p passenger sequence <400> 290 cuuaucaggu uguauuauaa uu 22 <210> 291 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-377-3p guide sequence <400> 291 aucacacaaa ggcaacuuuu gu 22 <210> 292 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-377-3p passenger sequence <400> 292 aaaaguugcc uuugugugau uu 22 <210> 293 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-378a-3p guide sequence <400> 293 acuggacuug gagucagaag gc 22 <210> 294 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-378a-3p passenger sequence <400> 294 cuucugacuc caaguccagu uu 22 <210> 295 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-383-5p guide sequence <400> 295 agaucagaag gugauugugg cu 22 <210> 296 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-383-5p passenger sequence <400> 296 ccacaaucac cuucugaucu uu 22 <210> 297 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-384 guide sequence <400> 297 auuccuagaa auuguucaua 20 <210> 298 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-384 passenger sequence <400> 298 ugaacaauuu cuaggaauuu 20 <210> 299 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-421 guide sequence <400> 299 aucaacagac auuaauuggg cgc 23 <210> 300 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-421 passenger sequence <400> 300 gcccaauuaa ugucuguuga uuu 23 <210> 301 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-422a guide sequence <400> 301 acuggacuua gggucagaag gc 22 <210> 302 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-422a passenger sequence <400> 302 cuucugaccc uaaguccagu uu 22 <210> 303 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-423-5p guide sequence <400> 303 ugaggggcag agagcgagac uuu 23 <210> 304 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-423-5p passenger sequence <400> 304 agucucgcuc ucugccccuc auu 23 <210> 305 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-424-5p guide sequence <400> 305 cagcagcaau ucauguuuug aa 22 <210> 306 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-424-5p passenger sequence <400> 306 caaaacauga auugcugcug uu 22 <210> 307 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-425-5p guide sequence <400> 307 aaugacacga ucacucccgu uga 23 <210> 308 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-425-5p passenger sequence <400> 308 aacgggagug aucgugucau uuu 23 <210> 309 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-429 guide sequence <400> 309 uaauacuguc ugguaaaacc gu 22 <210> 310 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-429 passenger sequence <400> 310 gguuuuacca gacaguauua uu 22 <210> 311 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-431-5p guide sequence <400> 311 ugucuugcag gccgucaugc a 21 <210> 312 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-431-5p passenger sequence <400> 312 caugacggcc ugcaagacau u 21 <210> 313 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-432-5p guide sequence <400> 313 ucuuggagua ggucauuggg ugg 23 <210> 314 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-432-5p passenger sequence <400> 314 acccaaugac cuacuccaag auu 23 <210> 315 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-448 guide sequence <400> 315 uugcauaugu aggauguccc au 22 <210> 316 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-448 passenger sequence <400> 316 gggacauccu acauaugcaa uu 22 <210> 317 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-449a guide sequence <400> 317 uggcagugua uuguuagcug gu 22 <210> 318 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-449a passenger sequence <400> 318 cagcuaacaa uacacugcca uu 22 <210> 319 <211> 25 <212> RNA <213> Artificial Sequence <220> <223> has-miR-449c-5p guide sequence <400> 319 uaggcagugu auugcuagcg gcugu 25 <210> 320 <211> 25 <212> RNA <213> Artificial Sequence <220> <223> has-miR-449c-5p passenger sequence <400> 320 agccgcuagc aauacacugc cuauu 25 <210> 321 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-450a-5p guide sequence <400> 321 uuuugcgaug uguuccuaau au 22 <210> 322 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-450a-5p passenger sequence <400> 322 auuaggaaca caucgcaaaa uu 22 <210> 323 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-450b-5p guide sequence <400> 323 uuuugcaaua uguuccugaa ua 22 <210> 324 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-450b-5p passenger sequence <400> 324 uucaggaaca uauugcaaaa uu 22 <210> 325 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-452-3p guide sequence <400> 325 cucaucugca aagaaguaag ug 22 <210> 326 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-452-3p passenger sequence <400> 326 cuuacuucuu ugcagaugag uu 22 <210> 327 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-486-5p guide sequence <400> 327 uccuguacug agcugccccg ag 22 <210> 328 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-486-5p passenger sequence <400> 328 cggggcagcu caguacagga uu 22 <210> 329 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-486-3p guide sequence <400> 329 cggggcagcu caguacagga u 21 <210> 330 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-mir-486-3p passenger sequence <400> 330 ccuguacuga gcugccccgu u 21 <210> 331 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-488-3p guide sequence <400> 331 uugaaaggcu auuucuuggu c 21 <210> 332 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-488-3p passenger sequence <400> 332 ccaagaaaua gccuuucaau u 21 <210> 333 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-489-3p guide sequence <400> 333 gugacaucac auauacggca gc 22 <210> 334 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-489-3p passenger sequence <400> 334 ugccguauau gugaugucac uu 22 <210> 335 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-491-5p guide sequence <400> 335 aguggggaac ccuuccauga gg 22 <210> 336 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-491-5p passenger sequence <400> 336 ucauggaagg guuccccacu uu 22 <210> 337 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-491-3p guide sequence <400> 337 cuuaugcaag auucccuucu ac 22 <210> 338 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-491-3p passenger sequence <400> 338 agaagggaau cuugcauaag uu 22 <210> 339 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-493-3p guide sequence <400> 339 ugaaggucua cugugugcca gg 22 <210> 340 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-493-3p passenger sequence <400> 340 uggcacacag uagaccuuca uu 22 <210> 341 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-494-3p guide sequence <400> 341 ugaaacauac acgggaaacc uc 22 <210> 342 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-494-3p passenger sequence <400> 342 gguuucccgu guauguuuca uu 22 <210> 343 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-495-3p guide sequence <400> 343 aaacaaacau ggugcacuuc uu 22 <210> 344 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-495-3p passenger sequence <400> 344 gaagugcacc auguuuguuu uu 22 <210> 345 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-497-5p guide sequence <400> 345 cagcagcaca cugugguuug u 21 <210> 346 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-497-5p passenger sequence <400> 346 aaaccacagu gugcugcugu u 21 <210> 347 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-498 guide sequence <400> 347 uuucaagcca gggggcguuu uuc 23 <210> 348 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-498 passenger sequence <400> 348 aaaacgcccc cuggcuugaa auu 23 <210> 349 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-499-5p guide sequence <400> 349 uuaagacuug cagugauguu u 21 <210> 350 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-499-5p passenger sequence <400> 350 acaucacugc aagucuuaau u 21 <210> 351 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-502-5p guide sequence <400> 351 auccuugcua ucugggugcu a 21 <210> 352 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-502-5p passenger sequence <400> 352 gcacccagau agcaaggauu u 21 <210> 353 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-502-3p guide sequence <400> 353 aaugcaccug ggcaaggauu ca 22 <210> 354 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-502-3p passenger sequence <400> 354 aauccuugcc caggugcauu uu 22 <210> 355 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-504-5p guide sequence <400> 355 agacccuggu cugcacucua uc 22 <210> 356 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-504-5p passenger sequence <400> 356 uagagugcag accagggucu uu 22 <210> 357 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-506-3p guide sequence <400> 357 uaaggcaccc uucugaguag a 21 <210> 358 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-506-3p passenger sequence <400> 358 uacucagaag ggugccuuau u 21 <210> 359 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-507 guide sequence <400> 359 uuuugcaccu uuuggaguga a 21 <210> 360 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-507 passenger sequence <400> 360 cacuccaaaa ggugcaaaau u 21 <210> 361 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-508-5p guide sequence <400> 361 uacuccagag ggcgucacuc aug 23 <210> 362 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-508-5p passenger sequence <400> 362 ugagugacgc ccucuggagu auu 23 <210> 363 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-509-5p guide sequence <400> 363 uacugcagac aguggcaauc a 21 <210> 364 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-509-5p passenger sequence <400> 364 auugccacug ucugcaguau u 21 <210> 365 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-510-5p guide sequence <400> 365 uacucaggag aguggcaauc ac 22 <210> 366 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-510-5p passenger sequence <400> 366 gauugccacu cuccugagua uu 22 <210> 367 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-511-5p guide sequence <400> 367 gugucuuuug cucugcaguc a 21 <210> 368 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-511-5p passenger sequence <400> 368 acugcagagc aaaagacacu u 21 <210> 369 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-512-3p guide sequence <400> 369 aagugcuguc auagcugagg uc 22 <210> 370 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-512-3p passenger sequence <400> 370 ccucagcuau gacagcacuu uu 22 <210> 371 <211> 18 <212> RNA <213> Artificial Sequence <220> <223> has-miR-513a-5p guide sequence <400> 371 uucacaggga ggugucau 18 <210> 372 <211> 18 <212> RNA <213> Artificial Sequence <220> <223> has-miR-513a-5p passenger sequence <400> 372 gacaccuccc ugugaauu 18 <210> 373 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-513a-3p guide sequence <400> 373 uaaauuucac cuuucugaga agg 23 <210> 374 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-513a-3p passenger sequence <400> 374 uucucagaaa ggugaaauuu auu 23 <210> 375 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-515-5p guide sequence <400> 375 uucuccaaaa gaaagcacuu ucug 24 <210> 376 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-515-5p passenger sequence <400> 376 gaaagugcuu ucuuuuggag aauu 24 <210> 377 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-516b-5p guide sequence <400> 377 aucuggaggu aagaagcacu uu 22 <210> 378 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-516b-5p passenger sequence <400> 378 agugcuucuu accuccagau uu 22 <210> 379 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-518a-5p guide sequence <400> 379 cucuagaggg aagcacuuuc ug 22 <210> 380 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-518a-5p passenger sequence <400> 380 gaaagugcuu cccucuagag uu 22 <210> 381 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-518a-3p guide sequence <400> 381 caaagcgcuu cccuuuggag c 21 <210> 382 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-518a-3p passenger sequence <400> 382 uccaaaggga agcgcuuugu u 21 <210> 383 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-518c-5p guide sequence <400> 383 ucucuggagg gaagcacuuu cug 23 <210> 384 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-518c-5p passenger sequence <400> 384 gaaagugcuu cccuccagag auu 23 <210> 385 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-mir-519d-3p guide sequence <400> 385 caaagugccu cccuuuagag ug 22 <210> 386 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-mir-519d-3p passenger sequence <400> 386 cucuaaaggg aggcacuuug uu 22 <210> 387 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-mir-519b-3p guide sequence <400> 387 aaagugcauc cuuuuagagg uu 22 <210> 388 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-mir-519b-3p passenger sequence <400> 388 ccucuaaaag gaugcacuuu uu 22 <210> 389 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-520a-5p guide sequence <400> 389 cuccagaggg aaguacuuuc u 21 <210> 390 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-520a-5p passenger sequence <400> 390 aaaguacuuc ccucuggagu u 21 <210> 391 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-520d-5p guide sequence <400> 391 cuacaaaggg aagcccuuuc 20 <210> 392 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-520d-5p passenger sequence <400> 392 aagggcuucc cuuuguaguu 20 <210> 393 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-520f-3p guide sequence <400> 393 aagugcuucc uuuuagaggg uu 22 <210> 394 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-520f-3p passenger sequence <400> 394 cccucuaaaa ggaagcacuu uu 22 <210> 395 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-522-3p guide sequence <400> 395 aaaaugguuc ccuuuagagu gu 22 <210> 396 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-522-3p passenger sequence <400> 396 acucuaaagg gaaccauuuu uu 22 <210> 397 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-525-5p guide sequence <400> 397 cuccagaggg augcacuuuc u 21 <210> 398 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-525-5p passenger sequence <400> 398 aaagugcauc ccucuggagu u 21 <210> 399 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-526a guide sequence <400> 399 cucuagaggg aagcacuuuc ug 22 <210> 400 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-526a passenger sequence <400> 400 gaaagugcuu cccucuagag uu 22 <210> 401 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-539-5p guide sequence <400> 401 ggagaaauua uccuuggugu gu 22 <210> 402 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-539-5p passenger sequence <400> 402 acaccaagga uaauuucucc uu 22 <210> 403 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-541-3p guide sequence <400> 403 uggugggcac agaaucugga cu 22 <210> 404 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-541-3p passenger sequence <400> 404 uccagauucu gugcccacca uu 22 <210> 405 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-543 guide sequence <400> 405 aaacauucgc ggugcacuuc uu 22 <210> 406 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-543 passenger sequence <400> 406 gaagugcacc gcgaauguuu uu 22 <210> 407 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-545-3p guide sequence <400> 407 ucagcaaaca uuuauugugu gc 22 <210> 408 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-545-3p passenger sequence <400> 408 acacaauaaa uguuugcuga uu 22 <210> 409 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-548b-5p guide sequence <400> 409 aaaaguaauu gugguuuugg cc 22 <210> 410 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-548b-5p passenger sequence <400> 410 ccaaaaccac aauuacuuuu uu 22 <210> 411 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-548b-3p guide sequence <400> 411 caagaaccuc aguugcuuuu gu 22 <210> 412 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-548b-3p passenger sequence <400> 412 aaaagcaacu gagguucuug uu 22 <210> 413 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-552-3p guide sequence <400> 413 aacaggugac ugguuagaca a 21 <210> 414 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-552-3p passenger sequence <400> 414 gucuaaccag ucaccuguuu u 21 <210> 415 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-556-3p guide sequence <400> 415 auauuaccau uagcucaucu uu 22 <210> 416 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-556-3p passenger sequence <400> 416 agaugagcua augguaauau uu 22 <210> 417 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-557 guide sequence <400> 417 guuugcacgg gugggccuug ucu 23 <210> 418 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-557 passenger sequence <400> 418 acaaggccca cccgugcaaa cuu 23 <210> 419 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-558 guide sequence <400> 419 ugagcugcug uaccaaaau 19 <210> 420 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-558 passenger sequence <400> 420 uuugguacag cagcucauu 19 <210> 421 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-559 guide sequence <400> 421 uaaaguaaau augcaccaaa a 21 <210> 422 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-559 passenger sequence <400> 422 uuggugcaua uuuacuuuau u 21 <210> 423 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-561-3p guide sequence <400> 423 caaaguuuaa gauccuugaa gu 22 <210> 424 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-561-3p passenger sequence <400> 424 uucaaggauc uuaaacuuug uu 22 <210> 425 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-568 guide sequence <400> 425 auguauaaau guauacacac 20 <210> 426 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-568 passenger sequence <400> 426 guguauacau uuauacauuu 20 <210> 427 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-570-3p guide sequence <400> 427 cgaaaacagc aauuaccuuu gc 22 <210> 428 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-570-3p passenger sequence <400> 428 aaagguaauu gcuguuuucg uu 22 <210> 429 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-571 guide sequence <400> 429 ugaguuggcc aucugaguga g 21 <210> 430 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-571 passenger sequence <400> 430 cacucagaug gccaacucau u 21 <210> 431 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-576-5p guide sequence <400> 431 auucuaauuu cuccacgucu uu 22 <210> 432 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-576-5p passenger sequence <400> 432 agacguggag aaauuagaau uu 22 <210> 433 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-576-3p guide sequence <400> 433 aagaugugga aaaauuggaa uc 22 <210> 434 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-576-3p passenger sequence <400> 434 uuccaauuuu uccacaucuu uu 22 <210> 435 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-577 guide sequence <400> 435 uagauaaaau auugguaccu g 21 <210> 436 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-577 passenger sequence <400> 436 gguaccaaua uuuuaucuau u 21 <210> 437 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-578 guide sequence <400> 437 cuucuugugc ucuaggauug u 21 <210> 438 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-578 passenger sequence <400> 438 aauccuagag cacaagaagu u 21 <210> 439 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-580-3p guide sequence <400> 439 uugagaauga ugaaucauua gg 22 <210> 440 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-580-3p passenger sequence <400> 440 uaaugauuca ucauucucaa uu 22 <210> 441 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-582-5p guide sequence <400> 441 uuacaguugu ucaaccaguu acu 23 <210> 442 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-582-5p passenger sequence <400> 442 uaacugguug aacaacugua auu 23 <210> 443 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-582-3p guide sequence <400> 443 uaacugguug aacaacugaa cc 22 <210> 444 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-582-3p passenger sequence <400> 444 uucaguuguu caaccaguua uu 22 <210> 445 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-584-5p guide sequence <400> 445 uuaugguuug ccugggacug ag 22 <210> 446 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-584-5p passenger sequence <400> 446 cagucccagg caaaccauaa uu 22 <210> 447 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-587 guide sequence <400> 447 uuuccauagg ugaugaguca c 21 <210> 448 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-587 passenger sequence <400> 448 gacucaucac cuauggaaau u 21 <210> 449 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-588 guide sequence <400> 449 uuggccacaa uggguuagaa c 21 <210> 450 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-588 passenger sequence <400> 450 ucuaacccau uguggccaau u 21 <210> 451 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-590-5p guide sequence <400> 451 gagcuuauuc auaaaagugc ag 22 <210> 452 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-590-5p passenger sequence <400> 452 gcacuuuuau gaauaagcuc uu 22 <210> 453 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-590-3p guide sequence <400> 453 uaauuuuaug uauaagcuag u 21 <210> 454 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-590-3p passenger sequence <400> 454 uagcuuauac auaaaauuau u 21 <210> 455 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-592 guide sequence <400> 455 uugugucaau augcgaugau gu 22 <210> 456 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-592 passenger sequence <400> 456 aucaucgcau auugacacaa uu 22 <210> 457 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-593-3p guide sequence <400> 457 ugucucugcu gggguuucu 19 <210> 458 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-593-3p passenger sequence <400> 458 aaaccccagc agagacauu 19 <210> 459 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-599 guide sequence <400> 459 guugugucag uuuaucaaac 20 <210> 460 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-599 passenger sequence <400> 460 uugauaaacu gacacaacuu 20 <210> 461 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-603 guide sequence <400> 461 cacacacugc aauuacuuuu gc 22 <210> 462 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-603 passenger sequence <400> 462 aaaaguaauu gcagugugug uu 22 <210> 463 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-605-5p guide sequence <400> 463 uaaaucccau ggugccuucu ccu 23 <210> 464 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-605-5p passenger sequence <400> 464 gagaaggcac caugggauuu auu 23 <210> 465 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-607 guide sequence <400> 465 guucaaaucc agaucuauaa c 21 <210> 466 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-607 passenger sequence <400> 466 uauagaucug gauuugaacu u 21 <210> 467 <211> 25 <212> RNA <213> Artificial Sequence <220> <223> has-miR-608 guide sequence <400> 467 agggguggug uugggacagc uccgu 25 <210> 468 <211> 25 <212> RNA <213> Artificial Sequence <220> <223> has-miR-608 passenger sequence <400> 468 ggagcugucc caacaccacc ccuuu 25 <210> 469 <211> 25 <212> RNA <213> Artificial Sequence <220> <223> has-miR-612 guide sequence <400> 469 gcugggcagg gcuucugagc uccuu 25 <210> 470 <211> 25 <212> RNA <213> Artificial Sequence <220> <223> has-miR-612 passenger sequence <400> 470 ggagcucaga agcccugccc agcuu 25 <210> 471 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-616-3p guide sequence <400> 471 agucauugga ggguuugagc ag 22 <210> 472 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-616-3p passenger sequence <400> 472 gcucaaaccc uccaaugacu uu 22 <210> 473 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-618 guide sequence <400> 473 aaacucuacu uguccuucug agu 23 <210> 474 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-618 passenger sequence <400> 474 ucagaaggac aaguagaguu uuu 23 <210> 475 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-619-3p guide sequence <400> 475 gaccuggaca uguuugugcc cagu 24 <210> 476 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-619-3p passenger sequence <400> 476 ugggcacaaa cauguccagg ucuu 24 <210> 477 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-623 guide sequence <400> 477 aucccuugca ggggcuguug ggu 23 <210> 478 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-623 passenger sequence <400> 478 ccaacagccc cugcaaggga uuu 23 <210> 479 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-624-3p guide sequence <400> 479 cacaagguau ugguauuacc u 21 <210> 480 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-624-3p passenger sequence <400> 480 guaauaccaa uaccuugugu u 21 <210> 481 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-626 guide sequence <400> 481 agcugucuga aaaugucuu 19 <210> 482 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-626 passenger sequence <400> 482 gacauuuuca gacagcuuu 19 <210> 483 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-635 guide sequence <400> 483 acuugggcac ugaaacaaug ucc 23 <210> 484 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-635 passenger sequence <400> 484 acauuguuuc agugcccaag uuu 23 <210> 485 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-636 guide sequence <400> 485 ugugcuugcu cgucccgccc gca 23 <210> 486 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-636 passenger sequence <400> 486 cgggcgggac gagcaagcac auu 23 <210> 487 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-637 guide sequence <400> 487 acugggggcu uucgggcucu gcgu 24 <210> 488 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-637 passenger sequence <400> 488 gcagagcccg aaagccccca guuu 24 <210> 489 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-641 guide sequence <400> 489 aaagacauag gauagaguca ccuc 24 <210> 490 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-641 passenger sequence <400> 490 ggugacucua uccuaugucu uuuu 24 <210> 491 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-642a-5p guide sequence <400> 491 gucccucucc aaaugugucu ug 22 <210> 492 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-642a-5p passenger sequence <400> 492 agacacauuu ggagagggac uu 22 <210> 493 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-642b-3p guide sequence <400> 493 agacacauuu ggagagggac cc 22 <210> 494 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-642b-3p passenger sequence <400> 494 gucccucucc aaaugugucu uu 22 <210> 495 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-643 guide sequence <400> 495 acuuguaugc uagcucaggu ag 22 <210> 496 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-643 passenger sequence <400> 496 accugagcua gcauacaagu uu 22 <210> 497 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-644a guide sequence <400> 497 aguguggcuu ucuuagagc 19 <210> 498 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-644a passenger sequence <400> 498 ucuaagaaag ccacacuuu 19 <210> 499 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-646 guide sequence <400> 499 aagcagcugc cucugaggc 19 <210> 500 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-646 passenger sequence <400> 500 cucagaggca gcugcuuuu 19 <210> 501 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-647 guide sequence <400> 501 guggcugcac ucacuuccuu c 21 <210> 502 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-647 passenger sequence <400> 502 aggaagugag ugcagccacu u 21 <210> 503 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-651-5p guide sequence <400> 503 uuuaggauaa gcuugacuuu ug 22 <210> 504 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-651-5p passenger sequence <400> 504 aaagucaagc uuauccuaaa uu 22 <210> 505 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-654-5p guide sequence <400> 505 uggugggccg cagaacaugu g 21 <210> 506 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-654-5p passenger sequence <400> 506 acauguucug cggcccacca uu 22 <210> 507 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-655-3p guide sequence <400> 507 auaauacaug guuaaccucu uu 22 <210> 508 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-655-3p passenger sequence <400> 508 agagguuaac cauguauuau uu 22 <210> 509 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-657 guide sequence <400> 509 ggcagguucu cacccucucu agg 23 <210> 510 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-657 passenger sequence <400> 510 uagagagggu gagaaccugc cuu 23 <210> 511 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-659-3p guide sequence <400> 511 cuugguucag ggaggguccc ca 22 <210> 512 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-659-3p passenger sequence <400> 512 gggacccucc cugaaccaag uu 22 <210> 513 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-661 guide sequence <400> 513 ugccuggguc ucuggccugc gcgu 24 <210> 514 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-661 passenger sequence <400> 514 gcgcaggcca gagacccagg cauu 24 <210> 515 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-663a guide sequence <400> 515 aggcggggcg ccgcgggacc gc 22 <210> 516 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-663a passenger sequence <400> 516 ggucccgcgg cgccccgccu uu 22 <210> 517 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-665 guide sequence <400> 517 accaggaggc ugaggccccu 20 <210> 518 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-665 passenger sequence <400> 518 gggccucagc cuccugguuu 20 <210> 519 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-671-5p guide sequence <400> 519 aggaagcccu ggaggggcug gag 23 <210> 520 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-671-5p passenger sequence <400> 520 ccagccccuc cagggcuucc uuu 23 <210> 521 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-708-5p guide sequence <400> 521 aaggagcuua caaucuagcu ggg 23 <210> 522 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-708-5p passenger sequence <400> 522 cagcuagauu guaagcuccu uuu 23 <210> 523 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-744-5p guide sequence <400> 523 ugcggggcua gggcuaacag ca 22 <210> 524 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-744-5p passenger sequence <400> 524 cuguuagccc uagccccgca uu 22 <210> 525 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-760 guide sequence <400> 525 cggcucuggg ucugugggga 20 <210> 526 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-760 passenger sequence <400> 526 cccacagacc cagagccguu 20 <210> 527 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-761 guide sequence <400> 527 gcagcagggu gaaacugaca ca 22 <210> 528 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-761 passenger sequence <400> 528 ugucaguuuc acccugcugc uu 22 <210> 529 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-765 guide sequence <400> 529 uggaggagaa ggaaggugau g 21 <210> 530 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-765 passenger sequence <400> 530 ucaccuuccu ucuccuccau u 21 <210> 531 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-767-5p guide sequence <400> 531 ugcaccaugg uugucugagc aug 23 <210> 532 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-767-5p passenger sequence <400> 532 ugcucagaca accauggugc auu 23 <210> 533 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-769-3p guide sequence <400> 533 cugggaucuc cggggucuug guu 23 <210> 534 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-769-3p passenger sequence <400> 534 ccaagacccc ggagauccca guu 23 <210> 535 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-770-5p guide sequence <400> 535 uccaguacca cgugucaggg cca 23 <210> 536 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-770-5p passenger sequence <400> 536 gcccugacac gugguacugg auu 23 <210> 537 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-802 guide sequence <400> 537 caguaacaaa gauucauccu ugu 23 <210> 538 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-802 passenger sequence <400> 538 aaggaugaau cuuuguuacu guu 23 <210> 539 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-874-3p guide sequence <400> 539 cugcccuggc ccgagggacc ga 22 <210> 540 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-874-3p passenger sequence <400> 540 ggucccucgg gccagggcag uu 22 <210> 541 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-875-3p guide sequence <400> 541 ccuggaaaca cugagguugu g 21 <210> 542 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-875-3p passenger sequence <400> 542 caaccucagu guuuccaggu u 21 <210> 543 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-876-5p guide sequence <400> 543 uggauuucuu ugugaaucac ca 22 <210> 544 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-876-5p passenger sequence <400> 544 gugauucaca aagaaaucca uu 22 <210> 545 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-876-3p guide sequence <400> 545 uggugguuua caaaguaauu ca 22 <210> 546 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-876-3p passenger sequence <400> 546 aauuacuuug uaaaccacca uu 22 <210> 547 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-877-5p guide sequence <400> 547 guagaggaga uggcgcaggg 20 <210> 548 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-877-5p passenger sequence <400> 548 cugcgccauc uccucuacuu 20 <210> 549 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-885-3p guide sequence <400> 549 aggcagcggg guguagugga ua 22 <210> 550 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-885-3p passenger sequence <400> 550 uccacuacac cccgcugccu uu 22 <210> 551 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-888-5p guide sequence <400> 551 uacucaaaaa gcugucaguc a 21 <210> 552 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-888-5p passenger sequence <400> 552 acugacagcu uuuugaguau u 21 <210> 553 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-922 guide sequence <400> 553 gcagcagaga auaggacuac guc 23 <210> 554 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-922 passenger sequence <400> 554 cguaguccua uucucugcug cuu 23 <210> 555 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-938 guide sequence <400> 555 ugcccuuaaa ggugaaccca gu 22 <210> 556 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-938 passenger sequence <400> 556 uggguucacc uuuaagggca uu 22 <210> 557 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-939-5p guide sequence <400> 557 uggggagcug aggcucuggg ggug 24 <210> 558 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-939-5p passenger sequence <400> 558 cccccagagc cucagcuccc cauu 24 <210> 559 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-940 guide sequence <400> 559 aaggcagggc ccccgcuccc c 21 <210> 560 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-940 passenger sequence <400> 560 ggagcggggg cccugccuuu u 21 <210> 561 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1178-3p guide sequence <400> 561 uugcucacug uucuucccua g 21 <210> 562 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1178-3p passenger sequence <400> 562 agggaagaac agugagcaau u 21 <210> 563 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1179 guide sequence <400> 563 aagcauucuu ucauugguug g 21 <210> 564 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1179 passenger sequence <400> 564 aaccaaugaa agaaugcuuu u 21 <210> 565 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1182 guide sequence <400> 565 gagggucuug ggagggaugu gac 23 <210> 566 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1182 passenger sequence <400> 566 cacaucccuc ccaagacccu cuu 23 <210> 567 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1197 guide sequence <400> 567 uaggacacau ggucuacuuc u 21 <210> 568 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1197 passenger sequence <400> 568 aaguagacca uguguccuau u 21 <210> 569 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1200 guide sequence <400> 569 cuccugagcc auucugagcc uc 22 <210> 570 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1200 passenger sequence <400> 570 ggcucagaau ggcucaggag uu 22 <210> 571 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1203 guide sequence <400> 571 cccggagcca ggaugcagcu c 21 <210> 572 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1203 passenger sequence <400> 572 gcugcauccu ggcuccgggu u 21 <210> 573 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1205 guide sequence <400> 573 ucugcagggu uugcuuugag 20 <210> 574 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1205 passenger sequence <400> 574 caaagcaaac ccugcagauu 20 <210> 575 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1206 guide sequence <400> 575 uguucaugua gauguuuaag c 21 <210> 576 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1206 passenger sequence <400> 576 uuaaacaucu acaugaacau u 21 <210> 577 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1207-5p guide sequence <400> 577 uggcagggag gcugggaggg g 21 <210> 578 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1207-5p passenger sequence <400> 578 ccucccagcc ucccugccau u 21 <210> 579 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1208 guide sequence <400> 579 ucacuguuca gacaggcgga 20 <210> 580 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1208 passenger sequence <400> 580 cgccugucug aacagugauu 20 <210> 581 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1224-5p guide sequence <400> 581 gugaggacuc gggaggugg 19 <210> 582 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1224-5p passenger sequence <400> 582 accucccgag uccucacuu 19 <210> 583 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1224-3p guide sequence <400> 583 ccccaccucc ucucuccuca g 21 <210> 584 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1224-3p passenger sequence <400> 584 gaggagagag gagguggggu u 21 <210> 585 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1228-3p guide sequence <400> 585 ucacaccugc cucgcccccc 20 <210> 586 <211> 20 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1228-3p passenger sequence <400> 586 ggggcgaggc aggugugauu 20 <210> 587 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1229-3p guide sequence <400> 587 cucucaccac ugcccuccca cag 23 <210> 588 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1229-3p passenger sequence <400> 588 gugggagggc aguggugaga guu 23 <210> 589 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1237-3p guide sequence <400> 589 uccuucugcu ccguccccca g 21 <210> 590 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1237-3p passenger sequence <400> 590 gggggacgga gcagaaggau u 21 <210> 591 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1246 guide sequence <400> 591 aauggauuuu uggagcagg 19 <210> 592 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1246 passenger sequence <400> 592 ugcuccaaaa auccauuuu 19 <210> 593 <211> 27 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1248 guide sequence <400> 593 accuucuugu auaagcacug ugcuaaa 27 <210> 594 <211> 27 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1248 passenger sequence <400> 594 uagcacagug cuuauacaag aagguuu 27 <210> 595 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1251-5p guide sequence <400> 595 acucuagcug ccaaaggcgc u 21 <210> 596 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1251-5p passenger sequence <400> 596 cgccuuuggc agcuagaguu u 21 <210> 597 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1254 guide sequence <400> 597 agccuggaag cuggagccug cagu 24 <210> 598 <211> 24 <212> RNA <213> Artificial Sequence <220> <223> has-miR-1254 passenger sequence <400> 598 ugcaggcucc agcuuccagg cuuu 24 <210> 599 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-let-7a-3p guide sequence <400> 599 cuauacaauc uacugucuuu c 21 <210> 600 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> hsa-let-7a-3p passenger sequence <400> 600 aagacaguag auuguauagu u 21 <210> 601 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> NC mimetics sense RNA <400> 601 uucuccgaac gugucacgut t 21 <210> 602 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> NC mimetics antisense RNA <400> 602 acgugacacg uucggagaat t 21 <210> 603 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-16-5p guide RNA <400> 603 uagcagcacg uaaauauugg cg 22 <210> 604 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-16-5p passenger RNA <400> 604 ccaauauuua cgugcugcua uu 22 <210> 605 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-542-3p guide RNA <400> 605 ugugacagau ugauaacuga aa 22 <210> 606 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-542-3p passenger RNA <400> 606 ucaguuauca aucugucaca uu 22 <210> 607 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> CCND1 siRNA Sense <400> 607 ggagaacaaa cagaucauct t 21 <210> 608 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> CCND1 siRNA Antisense <400> 608 gaugaucugu uuguucucct c 21 <210> 609 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> CCND3 siRNA Sense <400> 609 gggacagaau uggauacaut t 21 <210> 610 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> CCND3 siRNA Antisense <400> 610 auguauccaa uucuguccct t 21 <210> 611 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> CDK6 siRNA Sense <400> 611 gcacaaacau ucuggcauut t 21 <210> 612 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> CDK6 siRNA Antisense <400> 612 aaugccagaa uguuugugct t 21 <210> 613 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> Bcl2L1 siRNA Sense <400> 613 gccaucaaug gcaacccaut t 21 <210> 614 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> Bcl2L1 siRNA Antisense <400> 614 auggguugcc auugauggct t 21 <210> 615 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> Mcl1 siRNA Sense <400> 615 aaguaucaca gacguucuct t 21 <210> 616 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> Mcl1 siRNA Antisense <400> 616 gagaacgucu gugauacuut t 21 <210> 617 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> BIRC5 siRNA Sense <400> 617 aaggagauca acauuuucat t 21 <210> 618 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> BIRC5 siRNA Antisense <400> 618 ugaaaauguu gaucuccuut t 21 <210> 619 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> PIK3R2 siRNA Sense <400> 619 gcgaguauga ccagcuuuat t 21 <210> 620 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> PIK3R2 siRNA Antisense <400> 620 uaaagcuggu cauacucgct t 21 <210> 621 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> Sp1 siRNA Sense <400> 621 ccacuugccc uguucucuut t 21 <210> 622 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> Sp1 siRNA Antisense <400> 622 aagagaacag ggcaaguggt t 21 <210> 623 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> CYB5R3 siRNA Sense <400> 623 ggaacucagg aacaaacaut t 21 <210> 624 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> CYB5R3 siRNA Antisense <400> 624 auguuuguuc cugaguucct t 21

Claims (5)

A pharmaceutical composition for preventing or treating cancer comprising, as an active ingredient, miR-552-3p or a mimetic thereof, wherein the RNA having the nucleotide sequence of SEQ ID NO: 413 is combined with the RNA having the nucleotide sequence of SEQ ID NO: 414,
Wherein the cancer is lung cancer, liver cancer, or colon cancer.
The pharmaceutical composition for preventing or treating cancer according to claim 1, further comprising a pharmaceutically acceptable carrier.
The pharmaceutical composition according to claim 1, wherein said miRNA mimetic is contained in the form of a recombinant plasmid or viral vector expressing said miRNA mimetic or is contained together with a non-viral carrier.
The pharmaceutical composition for preventing or treating cancer according to claim 1, which is for promoting apoptosis or inhibiting cell growth.
The method according to claim 1, further comprising at least one active ingredient selected from the group consisting of an anti-cancer agent, a chemotherapeutic agent, an immunotherapeutic agent, an antibacterial agent, a radiotherapeutic agent and an antiviral agent, A pharmaceutical composition.

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