KR102371833B1 - MicroRNA for controlling expression of GDF11 gene and composition comprising the same - Google Patents

Abstract

The present invention relates to a microRNA controlling the expression of a growth differentiation factor 11 (GDF11) gene, and a composition containing the same. The microRNA-106b-5p, and microRNA-93-5p or microRNA-25-3p of the present invention can control the expression of the GDF11 gene, and thus are expected to be used for the treatment, prevention or control of diseases or physiological functions related to the GDF11 genes.

Description

MicroRNA for controlling expression of GDF11 gene and composition comprising the same

The present invention relates to a microRNA for regulating the expression of GDF11 gene and a composition containing the same.

Despite decades of efforts, cancer still ranks first or second among the causes of death among modern people due to disease. Such cancers are caused by genetic mutations caused by carcinogens or viruses in normal cells. The development and treatment of anticancer drugs target genes or proteins that are not expressed in normal cells but are specifically expressed in cancer cells. come. Anticancer agents are the most common and effective treatment methods for cancer treatment alone or in combination with other therapies such as radiotherapy. Treatment of cancer with these anticancer drugs is due to their ability to kill cancer cells, and it kills not only cancer cells but also normal cells, so side effects such as hair loss, anorexia, nausea, vomiting, dyspnea, stomatitis, neutropenic fever, etc. that it causes Depending on the patient's general health, such side effects may make anticancer drug treatment impossible or at least give the patient extreme discomfort and discomfort, and may seriously impair the cancer patient's quality of life.

MicroRNA (microRNA) is a short non-coding RNA consisting of about 18 to 25 nucleotides, and is generated by a hairpin-shaped transcript existing in a cell. MicroRNA binds to target mRNA (messenger RNA) complementarily and acts as a post-transcriptional gene repressor, and is known to inhibit gene expression by inhibiting mRNA translation or to induce destabilization by catalyzing mRNA cleavage. These microRNAs are involved in various roles in the body, such as apoptosis, cell cycle, and gene regulation, and by interaction with target genes, the overall organism including cancer differentiation, growth, metastasis, angiogenesis, and apoptosis. It can regulate the process, and the role and function of microRNAs in cancer can depend on the target gene.

On the other hand, Korean Patent No. 1042052 discloses 'a composition for preventing or treating solid cancer containing anti-microRNA' of miR-199a, which is specifically increased in production in cervical cancer, and Korean Patent No. 2110454 discloses ' Although the 'composition for preventing or treating cancer diseases comprising microRNA-550a-3-5p and an anticancer agent as active ingredients' is disclosed, the microRNA regulating the expression of the GDF11 gene of the present invention and a composition containing the same have not been described.

The present invention was derived from the above needs, and the present inventors confirmed that the target sequences of microRNA-106b-5p, microRNA-93-5p and microRNA-25-3p exist in GDF11 mRNA, and the microRNA-106b By overexpressing -5p, microRNA-93-5p and microRNA-25-3p, it was confirmed that the expression level of the GDF11 gene was reduced, thereby completing the present invention.

In order to solve the above problems, the present invention is microRNA-106b-5p consisting of the nucleotide sequence of SEQ ID NO: 1, microRNA-93-5p consisting of the nucleotide sequence of SEQ ID NO: 2, and microRNA-25- consisting of the nucleotide sequence of SEQ ID NO: 3 It provides a composition for inhibiting the expression of GDF11 (Growth differentiation factor 11) gene comprising one or more microRNAs selected from the group consisting of 3p as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer comprising one or more microRNAs selected from the group consisting of microRNA-106b-5p, microRNA-93-5p and microRNA-25-3p as an active ingredient.

In addition, the present invention provides an anticancer adjuvant for the prevention or treatment of cancer comprising one or more microRNAs selected from the group consisting of microRNA-106b-5p, microRNA-93-5p and microRNA-25-3p as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer comprising an inhibitor of expression of one or more microRNAs selected from the group consisting of microRNA-106b-5p, microRNA-93-5p and microRNA-25-3p as an active ingredient do.

Since the microRNA-106b-5p, microRNA-93-5p or microRNA-25-3p of the present invention can regulate the expression of the GDF11 gene, it can be used for the treatment, prevention or regulation of diseases or physiological functions related to the overexpression of the GDF11 gene. will be.

1A is the result of measuring the cell viability of the human liver cancer cell line Huh7 transformed with Mock, GDF11 expression vector, p53 expression vector, or Mutant p53 expression vector, and FIG. 1B is Mock, GDF11 expression vector, p53 expression vector or Mutant p53 expression vector This is the result of confirming the colony forming ability of Huh7 cells transformed with the vector. Mock: Huh7 cells untreated.
2A is a diagram showing clustered primary microRNA-106b-25 isolated from human primary microRNA and microRNA-106b-5p, microRNA- 93-5p and microRNA-25-3p generated therefrom, and FIG. These are the results of confirming the target sequences of microRNA-106b-5p, microRNA-93-5p and microRNA-25-3p in the 3'-UTR region.
3A to 3C show when overexpressing clustered primary microRNA-106b-25 isolated from human primary microRNA in Huh7 cells, GDF11 protein or its Western blot (A), qRT-PCR (Quantitative real-time PCR; B) and Luciferase reporter assay confirming that the expression level of the gene is reduced compared to the control (empty or wild type) ; C) is the result. empty: (psiCheck-2 vector treatment group), wild type: Among the 3'-UTR sequences of GDF11 mRNA, about 1,857 bp sequences including the target sequences of microRNA-106b-5p, microRNA-93-5p and microRNA-25-3p were cloned into the psiCheck-2 vector and then expressed, miRNA target site mutant: Mutating the target sequences of microRNA-106b-5p, microRNA-93-5p and microRNA-25-3p among the 3'-UTR sequences of GDF11 mRNA (the target sequence of microRNA-25-3p among the GDF11 3'-UTR sequences GUGCAAU was converted to GUGAAUC, and GCACUUU, the target sequence of microRNA-93-5p and microRNA-106b-5p, was converted to GCUAUUC). The sequence of about 1,857 bp was cloned into the psiCheck-2 vector and expressed.
Figure 4A shows that the expression level of the GDF11 gene decreases compared to the control (wild type) after microRNA-93-5p or microRNA-25-3p itself generated from clustered primary microRNA-106b-25 is treated in Huh7 cells, respectively. Results of a luciferase reporter assay, and FIG. 4B is a result of a luciferase reporter assay confirming the expression level of the GDF11 gene in Huh7 cells treated with microRNA-93-5p inhibitor (anti-miR-93). WT: Mock (untreated Huh7 cells), anti-miR-93: microRNA-93-5p activity inhibitor (a complementary sequence capable of targeting microRNA-93-5p, all nucleotide 2 hydroxyl groups are 2 Including the sequence modified with '-O-methyl), anti-siGFP: negative control (sequence in which the 2nd hydroxyl group of all nucleotides of RNA, which is not expected to have any effect on cells, is modified with 2'-O-methyl including).

In order to achieve the object of the present invention, the present invention is microRNA-106b-5p consisting of the nucleotide sequence of SEQ ID NO: 1, microRNA-93-5p consisting of the nucleotide sequence of SEQ ID NO: 2, and microRNA- It provides a composition for inhibiting the expression of GDF11 (Growth differentiation factor 11) gene comprising one or more microRNAs selected from the group consisting of 25-3p as an active ingredient.

In the composition of the present invention, the microRNA-106b-5p, microRNA-93-5p and microRNA-25-3p may bind to the 3'-UTR region of GDF11 mRNA, and the microRNA-106b-5p, microRNA-93 When -5p or microRNA-25-3p is overexpressed in cancer cells, it has the effect of suppressing the expression of the GDF11 gene.

The present invention also, from the group consisting of microRNA-106b-5p consisting of the nucleotide sequence of SEQ ID NO: 1, microRNA-93-5p consisting of the nucleotide sequence of SEQ ID NO: 2, and microRNA-25-3p consisting of the nucleotide sequence of SEQ ID NO: 3 It provides a pharmaceutical composition for preventing or treating cancer comprising one or more selected microRNAs as an active ingredient.

As used herein, the term "cancer" refers to general cancer diseases including solid tumors and blood born tumors, preferably liver cancer, lung cancer, stomach cancer, colon cancer, breast cancer, and non-small cell lung cancer. , bone cancer, pancreatic cancer, skin cancer, head or cervical cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, colorectal cancer, small intestine cancer, rectal cancer, perianal cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulva Carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, lymph gland cancer, bladder cancer, gallbladder cancer, endocrine adenocarcinoma, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia; lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem glioma, pituitary adenoma or fibrosarcoma cancer; Preferably, it may be liver cancer, but is not limited thereto.

In the pharmaceutical composition of the present invention, by regulating the expression of the microRNA-106b-5p, microRNA-93-5p or microRNA-25-3p, the proliferation of cancer cells is controlled by regulating the expression of the GDF11 (Growth differentiation factor 11) gene. Preferably, by increasing the expression of the microRNA-106b-5p, microRNA-93-5p or microRNA-25-3p in cancer cells to decrease the expression of the GDF11 gene, thereby increasing the proliferation of cancer cells. Alternatively, by inhibiting the expression of the microRNA-106b-5p, microRNA-93-5p or microRNA-25-3p to increase the expression of the GDF11 gene, it may inhibit the proliferation of cancer cells, but is not limited thereto.

The pharmaceutical dosage form of the composition according to the present invention may be used alone or in combination with other pharmaceutically active compounds, as well as in any suitable group.

The pharmaceutical composition according to the present invention may be formulated in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions according to conventional methods, respectively. may be, but is not limited thereto. Carriers, excipients and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose ) or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral use include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, Witepsol, macrogol, Tween 61, cacao butter, laurin fat, glycerol, geratin, etc. may be used.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and when administered parenterally, it is preferable to select an external skin or intraperitoneal, rectal, intravenous, muscle, subcutaneous, intrauterine dural or intracerebrovascular injection method, but limited thereto. I never do that.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, a pharmaceutically effective amount means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level depends on the type, severity, drug activity, and drug of the patient. Sensitivity, administration time, administration route and excretion rate, treatment duration, factors including concurrent drugs, and other factors well known in the medical field may be determined. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.

The dosage of the composition of the present invention may be administered in various ways depending on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate and severity of disease, but administration route, severity of obesity , gender, weight, age, etc. may be increased or decreased according to the dosage, it is not intended to limit the scope of the present invention in any way.

The present invention also, from the group consisting of microRNA-106b-5p consisting of the nucleotide sequence of SEQ ID NO: 1, microRNA-93-5p consisting of the nucleotide sequence of SEQ ID NO: 2, and microRNA-25-3p consisting of the nucleotide sequence of SEQ ID NO: 3 It provides an anticancer adjuvant for the prevention or treatment of cancer comprising one or more selected microRNAs as an active ingredient.

The present invention also, from the group consisting of microRNA-106b-5p consisting of the nucleotide sequence of SEQ ID NO: 1, microRNA-93-5p consisting of the nucleotide sequence of SEQ ID NO: 2, and microRNA-25-3p consisting of the nucleotide sequence of SEQ ID NO: 3 It provides a pharmaceutical composition for the prevention or treatment of cancer comprising one or more selected microRNA expression inhibitors as an active ingredient.

The microRNA expression inhibitor can be used as a preventive or therapeutic agent for cancer by inhibiting the proliferation of cancer cells by increasing the expression of the GDF11 gene by inhibiting the expression of the microRNA.

Hereinafter, the present invention will be described in detail by way of Examples. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples.

Materials and Methods

1. Cell Culture and Plasmid Construction

The human liver cancer cell line Huh7 was cultured using DMEM (Dulbecco's modified eagle's medium) medium containing 9% fetal bovine serum.

Huh7 was transformed by constructing a GDF11 expression vector, a p53 expression vector, or a Mutant p53 expression vector for cell viability and cell colony formation analysis. Specifically, the GDF11 gene and p53 gene were amplified from HEK293T cDNA and inserted into pcDNA3.1 vector using an overlap cloner DNA cloning kit. induced to have Each of the cloned plasmids, anti-siGFP or anti-miR-93 (Bioneer) was transformed into Huh7 cells according to the manufacturer's method of the Lipofectamine 2000 transfection reagent (Thermofisher scientific). The anti-miR-93 is a single-stranded synthetic inhibitor having a complementary sequence so as to target microRNA-93-5p, and all nucleotide 2 hydroxyl groups are modified to 2'-O-methyl to microRNA-93-5p anti-siGFP, which is expected to have no effect on cells, is a 2'-O-methyl was used as a negative control.

In addition, microRNA-106b-5p (SEQ ID NO: 1) isolated from human primary microRNA to perform Western blot, quantitative real-time PCR (qRT-PCR) and Luciferase reporter assay. : 5'-UAAAGUGCUGACAGUGCAGAU-3'), microRNA-93-5p (SEQ ID NO: 2: 5'-CAAAAGUGCUGUUCGUGCAGGUAG-3'), microRNA-25-3p (SEQ ID NO: 3: 5'CAUUGCACUUGUCUCGGUCUGA-3') or clustered primary microRNA -106b-25 (SEQ ID NO: 4) was amplified from the genomic DNA of HEK293T (Human embryonic kidney 293 T) cells and cloned into pcDNA3.1 vector, respectively. For the luciferase reporter assay, GDF11 3' untranslated sequence (1,296-3,152 bp) was amplified from HeLa cell cDNA and cloned into psicheck-2 vector (Promega), and site-specific mutation of the GDF11 3' untranslated sequence Induction was carried out through inverted PCR. All cloning processes were performed using an Overlap cloner DNA cloning kit (Elpis biotech.).

2. Measurement of Cell Viability

After culturing the transformed Huh7 cells for 48 hours, 2×10 ³ cells/100 μl were inoculated into each well of a 96-well plate and cultured for a total of 4 days in a cell incubator at 37° C., every day at intervals of 1 day. At the same time, the cells were treated with MTS/PMS (Promega) solution and reacted for 60 minutes, then cell viability was measured using CellTiter 96 AQueous Non-Radioactive Cell Proliferation Assay (Promega).

3. colony formation assay

In order to measure the adhesion-independent colony forming ability of cells, a soft agar assay was performed. Specifically, a 6-well plate is treated with 0.6% agarose gel to harden to form a relatively hard lower layer, and then treated with 0.3% agarose gel containing 1×10 ⁴ cells and cell culture solution on it to harden, and then approximately twice a week The culture medium was treated and cultured for 3 weeks, and after staining with crystal violet, the number of colonies was measured using image software (ImageJ).

4. Western blot

The transformed Huh7 cells were treated with a RIPA (Radioimmunoprecipitation assay) buffer, followed by repeated vortexing and centrifugation at 12,000 rpm to obtain a cell lysate. After that, a total of 30 μg of protein was loaded on a 10% polyacrylamide gel, separated, and transferred to a PVDF membrane. For GDF11 detection, Rabbit anti-GDF11 polyclonal antibody (Abcam, ab71347) as a primary antibody, and Rabbit anti-GAPDH monoclonal antibody (Cell Signaling Technology, #2118) as a primary antibody for GAPDH detection were used. As the secondary antibody, Goat anti-rabbit IgG (H+L) HRP-conjugated antibody was used. After the antibody-treated membrane was reacted with ECL reagent (Elpis biotech.), it was exposed to an X-ray film for analysis.

5. Quantitative real-time PCR (qRT-PCR)

After treating the transformed Huh7 cells with RibeEx (GeneAll), RNA was extracted and purified through an RNA isolation method using phenol/chloroform, and DNase I was additionally treated to prevent DNA contamination. Finally, cDNA was synthesized from the purified RNA using Oligo d(T) and PrimeScript reverse transcriptase (Takara). Relative gene expression levels were measured using synthesized cDNA and LightCycler ^® 480 SYBR Green I Master (Roche Life Science), and LightCycler 480 Instrument II (Roche Life Science).

6. Luciferase reporter assay

The luciferase reporter assay was performed using the Dual-luciferase ^® reporter assay system (Promega). Specifically, Huh7 cells were inoculated into a 24-well plate at 1×10 ⁴ cells per well, and 48 hours after transformation, the transformed cells were lysed by treatment with passive lysis buffer (Promega), and luciferase assay reagent II ( LAR II) was treated to measure firefly luciferase activity, and then Stop & Glo ^® Buffer was re-treated to measure Renilla luciferase activity. Renilla luciferase and firefly luciferase activities were respectively measured using a Glomax ^® 96 microplate luminometer (Promega). Analysis was normalized to activity.

Example 1. GDF11 Analysis of the anticancer effect of genes

To analyze the effect of the GDF11 gene on the proliferation of liver cancer cells, cell viability and colony formation assay were analyzed after overexpressing the GDF11 gene in the human liver cancer cell line Huh7.

As a result of measuring the cell viability, it was confirmed that the cell viability was significantly reduced in the GDF11 overexpression test group compared to the control group (Mock), and at a level similar to the p53 overexpression test group known as an anticancer gene (FIG. 1A).

In addition, as a result of measuring the cell colony forming ability, it was confirmed that the cell population was significantly reduced in the GDF11 overexpression test group compared to the control group (Mock), and the colony was formed at a level similar to that of the p53 overexpression test group (FIG. 1B). Through this, it was found that the GDF11 gene has anticancer activity against liver cancer.

Example 2. GDF11 Analysis of microRNA target sequences in mRNA

Target sites of microRNA-106b-5p, microRNA-93-5p, and microRNA-25-3p in GDF11 mRNA were identified using the microRNA target prediction program TargetScan.

As a result, assuming that 1 bp immediately after UAA, the stop codon, in the CDS (coding sequence) of GDF11 mRNA (GenBank accession No: NM_005811.5) is 1 bp, 130-136 of GDF11 3'-UTR A partial sequence of microRNA-25-3p (AUUGCAC) can bind to the bp region, and a partial sequence (AAAGUGC) of microRNA-106b-5p can bind to the 1,626-1,632 bp region and 3,639-3,645 bp region, and 1,626 It was confirmed that some sequences of microRNA-93-5p (AAAGUGC) can bind to ~1,632 bp region and 3,639 ~ 3,645 bp region (FIG. 2B). That is, since the target sequences of microRNA-106b-5p, microRNA-93-5p, and microRNA-25-3p exist in GDF11 mRNA, it was thought that the three microRNAs could regulate the expression of the GDF11 gene.

Example 3. Through regulation of microRNA expression GDF11 gene expression regulation analysis

To analyze the effect of microRNA-106b-5p, microRNA-93-5p and microRNA-25-3p of the present invention on the expression level of GDF11 gene, clustered primary microRNA-106b-25 isolated from human primary microRNA in Huh7 cells After overexpression, Western blot, qRT-PCR and luciferase reporter assay were performed to analyze the expression level of the GDF11 gene.

As a result, it was confirmed that when clustered primary microRNA-106b-25 was simultaneously overexpressed in Huh7 cells, the expression of the GDF11 gene was significantly reduced compared to the control group (empty) ( FIGS. 3A and 3B ). In addition, a luciferase reporter assay was performed on the clustered primary microRNA-106b-25 treatment group and mutants having a sequence in which the target sequence of microRNA-106b-25 in the GDF11 gene was arbitrarily changed, and microRNA-106b-25 By confirming that the inhibition rate of GDF11 expression in the treatment group (about 2.2 fold) was significantly higher than that of the miRNA target site mutant treatment group (about 1.0 fold), the specific sequence of microRNA-106b-25 targeting the GDF11 gene in the present invention actually It was found that the expression of this GDF11 gene can be regulated (FIG. 3C).

In addition, as a result of analyzing the effect of microRNA-93-5p or microRNA-25-3p generated in clustered primary microRNA-106b-25 on the expression level of GDF11 gene, microRNA-93-5p or microRNA-25-3p was When expressed in cells, it was confirmed that the suppression effect of the GDF11 gene was increased (FIG. 4A).

In addition, in order to check the effect of microRNA activity inhibition on the expression of the GDF11 gene, Huh7 cells were treated with a microRNA-93-5 inhibitor (anti-miR-93) and then the expression level of the GDF11 gene was analyzed. anti-siGFP), it was confirmed that the suppression effect of the GDF11 gene was reduced in the microRNA-93-5 inhibitor-treated group (FIG. 4B).

<110> Seoul National University R&DB Foundation <120> MicroRNA for controlling expression of GDF11 gene and composition comprising the same <130> PN20215 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> RNA <213> Artificial Sequence <220> <223> microRNA-106b-5p <400> 1 uaaagugcug acagugcaga u 21 <210> 2 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> microRNA-93-5p <400> 2 caaagugcug uucgugcagg uag 23 <210> 3 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> microRNA-25-3p <400> 3 cauugcacuu gucucggucu ga 22 <210> 4 <211> 515 <212> RNA <213> Artificial Sequence <220> <223> clustered primary microRNA-106b~25 <400> 4 cctgccgggg ctaaagtgct gacagtgcag atagtggtcc tctccgtgct accgcactgt 60 gggtacttgc tgctccagca gggcacgcac agcgtccgtg gagggaaagg ccttttcccc 120 acttcttaac cttcactgag agggtggttg gggtctgttt cactccatgt gtcctagatc 180 ctgtgctaca gaccttcctt tctgtcctcc cgtcttggac ctcagtcctg ggggctccaa 240 agtgctgttc gtgcaggtag tgtgattacc caacctactg ctgagctagc acttcccgag 300 cccccgggac acgttctctc tgccaattgt cttcttggct gagctcccca agctccatct 360 gtcatgctgg ggagcccagt ggcgttcaaa agggtctggt ctccctcaca ggacagctga 420 actccgggac tggccagtgt tgagaggcgg agacttgggc aattgctgga cgctgccctg 480 ggcattgcac ttgtctcggt ctgacagtgc cggcc 515

Claims (5)

One or more microRNAs selected from the group consisting of microRNA-106b-5p consisting of the nucleotide sequence of SEQ ID NO: 1, microRNA-93-5p consisting of the nucleotide sequence of SEQ ID NO: 2, and microRNA-25-3p consisting of the nucleotide sequence of SEQ ID NO: 3 A composition for inhibiting the expression of GDF11 (Growth differentiation factor 11) gene comprising as an active ingredient. delete delete delete delete

Images