Expression of miR-409-3p in hepatoma HepG2 cells and its mechanism in cell proliferation

Changqing WANG; Ling CHEN; Ping XU; Xiaojuan ZHU; Zheng LIU

doi:10.3969/j.issn.1001-5256.2023.08.019

Volume 39 Issue 8

Aug. 2023

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Article Navigation > Journal of Clinical Hepatology > 2023 > 39(8): 1895-1902

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Expression of miR-409-3p in hepatoma HepG2 cells and its mechanism in cell proliferation

DOI: 10.3969/j.issn.1001-5256.2023.08.019

1.
Department of Gastroenterology, Yancheng First People's Hospital, Yancheng, Jiangsu 224008, China
2.
Department of Gastroenterology, Nanjing Pukou People's Hospital, Nanjing 211899, China
3.
Medical Center for Digestive Diseases, The Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, China

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Corresponding author: LIU Zheng, liuzheng117@126.com (ORCID: 0000-0002-2195-4538)
Received Date: 2022-11-05
Accepted Date: 2023-02-27
Published Date: 2023-08-20

Abstract

Abstract

Objective To investigate the expression and significance of miR-409-3p in hepatoma carcinoma cell lines and possible molecular mechanism. Methods Quantitative real-time PCR was used to measure the expression of miR-409-3p in normal LO2 hepatocytes and four hepatoma cell line (HepG2, BEL-7402, SMMC-7721, and MHCC-97H). Hepatoma HepG2 cells were transiently transfected with miR-409-3p mimics and microRNA mimics control using the cationic liposome method, and then CCK8, plate colony formation assay, and flow cytometry were used to observe the effect of miR-409-3p on the proliferation, cell cycle, and apoptosis of hepatoma cells in vitro. Western blot was used to measure the change in the expression of c-Met protein in HepG2 cells with overexpression of miR-409-3p, and luciferase reporter gene assay was used to identify targeting relationship. The independent-samples t test was used for comparison between two groups, and a one-way analysis of variance was used for comparison between multiple groups, followed by the SNK test. Results Based on the TCGA microRNA expression profile data of liver cancer, the expression level of miR-409-3p in liver cancer tissue was significantly lower than that in adjacent tissue (t=7.752, P < 0.05). Compared with the LO2 cells, the HepG2, SMMC-7721, MHCC-97H, and BEL-7402 cells had a significant reduction in the expression level of miR-409-3p (F=31.043, P < 0.05). Compared with the miR-con group, the HepG2 cells transfected with miR-409-3p mimics had a significant increase in the expression level of miR-409-3p (t=-8.836, P < 0.05), suggesting that the interference was effective. CCK8 assay showed that compared with the miR-con group, the HepG2 cells transfected with miR-409-3p mimics had a significant reduction in proliferative capacity at 48, 72, and 96 hours (t=2.876, 3.359, and 3.707, all P < 0.05). Plate colony formation assay showed that the miR-409-3p mimics group had a significantly lower plating efficiency than the miR-con group (t=2.846, P=0.047). Flow cytometry showed that compared with the miR-con group, overexpression of miR-409-3p resulted in the increased number of HepG2 cells in G2 phase (t=-3.763, P < 0.05), while there was no significant difference in apoptosis rate (t=0.714, P=0.515). Luciferase reporter gene assay showed that c-Met was a target gene of miR-409-3p (t=4.970, P=0.007). Compared with the miR-con group, the HepG2 cells transfected with miR-409-3p mimics had a significant reduction in the expression of c-Met protein (t=-8.509, P=0.001). Conclusion By inhibiting the protein expression of c-Met, miR-409-3p regulates downstream signaling pathways to induce cell cycle arrest in G2 phase and thus inhibits the proliferation of hepatoma HepG2 cells.
- Carcinoma, Hepatocellular,
- MicroRNAs,
- Proto-Oncogene Proteins c-met,
- Cell Proliferation

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References

[1]	RUIZ-MANRIQUEZ LM, CARRASCO-MORALES O, SANCHEZ Z EA, et al. MicroRNA-mediated regulation of key signaling pathways in hepatocellular carcinoma: A mechanistic insight[J]. Front Genet, 2022, 13: 910733. DOI: 10.3389/fgene.2022.910733.
[2]	COOPER CS, PARK M, BLAIR DG, et al. Molecular cloning of a new transforming gene from a chemically transformed human cell line[J]. Nature, 1984, 311(5981): 29-33. DOI: 10.1038/311029a0.
[3]	FAIELLA A, RICCARDI F, CARTENÌ G, et al. The emerging role of c-Met in carcinogenesis and clinical implications as a possible therapeutic target[J]. J Oncol, 2022, 2022: 5179182. DOI: 10.1155/2022/5179182.
[4]	CHEN L, SHI Y, ZHU X, et al. IL-10 secreted by cancer-associated macrophages regulates proliferation and invasion in gastric cancer cells via c-Met/STAT3 signaling[J]. Oncol Rep, 2019, 42(2): 595-604. DOI: 10.3892/or.2019.7206.
[5]	MA Y, ZHANG M, WANG J, et al. High-affinity human anti-c-Met IgG conjugated to oxaliplatin as targeted chemotherapy for hepatocellular carcinoma[J]. Front Oncol, 2019, 9: 717. DOI: 10.3389/fonc.2019.00717.
[6]	XU X, CHEN H, LIN Y, et al. MicroRNA-409-3p inhibits migration and invasion of bladder cancer cells via targeting c-Met[J]. Mol Cells, 2013, 36(1): 62-68. DOI: 10.1007/s10059-013-0044-7.
[7]	WAN L, ZHU L, XU J, et al. MicroRNA-409-3p functions as a tumor suppressor in human lung adenocarcinoma by targeting c-Met[J]. Cell Physiol Biochem, 2014, 34(4): 1273-1290. DOI: 10.1159/000366337.
[8]	General Office of National Health Commission. Standard for diagnosis and treatment of primary liver cancer (2022 edition)[J]. J Clin Hepatol, 2022, 38(2): 288-303. DOI: 10.3969/j.issn.1001-5256.2022.02.009. 国家卫生健康委办公厅. 原发性肝癌诊疗指南(2022年版)[J]. 临床肝胆病杂志, 2022, 38(2): 288-303. DOI: 10.3969/j.issn.1001-5256.2022.02.009.
[9]	SIEGEL RL, MILLER KD, GODING SAUER A, et al. Colorectal cancer statistics, 2020[J]. CA Cancer J Clin, 2020, 70(3): 145-164. DOI: 10.3322/caac.21601.
[10]	FORNER A, LLOVET JM, BRUIX J. Hepatocellular carcinoma[J]. Lancet, 2012, 379(9822): 1245-1255. DOI: 10.1016/S0140-6736(11)61347-0.
[11]	RAY K. Liver cancer: The promise of new approaches in the management of hepatocellular carcinoma--adding to the toolbox?[J]. Nat Rev Gastroenterol Hepatol, 2013, 10(4): 195. DOI: 10.1038/nrgastro.2013.52.
[12]	TRINCHET JC, CHAFFAUT C, BOURCIER V, et al. Ultrasonographic surveillance of hepatocellular carcinoma in cirrhosis: a randomized trial comparing 3- and 6-month periodicities[J]. Hepatology, 2011, 54(6): 1987-1997. DOI: 10.1002/hep.24545.
[13]	ZHOU Q, SHAO JG. Research progress of miRNA in HBV-related hepatocellular carcinoma[J]. J Nantong Univ(Med Sci), 2022, 42(3): 257-261. DOI: 10.16424/j.cnki.cn32-1807/r.2022.03.013. 周倩, 邵建国. MiRNA在HBV相关肝癌中的研究进展[J]. 南通大学学报(医学版), 2022, 42(3): 257-261. DOI: 10.16424/j.cnki.cn32-1807/r.2022.03.013.
[14]	XIE HJ, RASHED N, NING Y, et al. Current status of research on circulating microRNAs as diagnostic markers for hepatocellular carcinoma[J]. J Clin Hepatol, 2021, 37(2): 448-451. DOI: 10.3969/j.issn.1001-5256.2021.02.042. 谢惠君, Rashed Nasot, 宁勇, 等. 循环miRNA作为肝细胞癌标志物的研究现状[J]. 临床肝胆病杂志, 2021, 37(2): 448-451. DOI: 10.3969/j.issn.1001-5256.2021.02.042.
[15]	HUSSEN BM, HIDAYAT HJ, SALIHI A, et al. MicroRNA: A signature for cancer progression[J]. Biomed Pharmacother, 2021, 138: 111528. DOI: 10.1016/j.biopha.2021.111528.
[16]	PIEROULI K, PAPAKONSTANTINOU E, PAPAGEORGIOU L, et al. Long non-coding RNAs and microRNAs as regulators of stress in cancer (Review)[J]. Mol Med Rep, 2022, 26(6): 361. DOI: 10.3892/mmr.2022.12878.
[17]	HUANG S, HE X. The role of microRNAs in liver cancer progression[J]. Br J Cancer, 2011, 104(2): 235-240. DOI: 10.1038/sj.bjc.6606010.
[18]	LIU S, LI B, XU J, et al. SOD1 Promotes cell proliferation and metastasis in non-small cell lung cancer via an miR-409-3p/SOD1/SETDB1 epigenetic regulatory feedforward loop[J]. Front Cell Dev Biol, 2020, 8: 213. DOI: 10.3389/fcell.2020.00213.
[19]	WANG Y, ZHANG J, CHEN X, et al. Circ_0001023 promotes proliferation and metastasis of gastric cancer cells thro ugh miR-409-3p/PHF10 axis[J]. Onco Targets Ther, 2020, 13: 4533-4544. DOI: 10.2147/OTT.S244358.
[20]	CUI X, CHEN J, ZHENG Y, et al. Circ_0000745 promotes the progression of cervical cancer by regulating miR-409-3p/ATF1 axis[J]. Cancer Biother Radiopharm, 2022, 37(9): 766-778. DOI: 10.1089/cbr.2019.3392.
[21]	CHEN J, WANG R, LU E, et al. LINC00630 as a miR-409-3p sponge promotes apoptosis and glycolysis of colon carcinoma cells via regulating HK2[J]. Am J Transl Res, 2022, 14(2): 863-875.
[22]	YANG S, ZOU C, LI Y, et al. Knockdown circTRIM28 enhances tamoxifen sensitivity via the miR-409-3p/HMGA2 axis in breast cancer[J]. Reprod Biol Endocrinol, 2022, 20(1): 146. DOI: 10.1186/s12958-022-01011-3.
[23]	KOTA J, CHIVUKULA RR, O'DONNELL KA, et al. Therapeutic microRNA delivery suppresses tumorigenesis in a murine liver cancer model[J]. Cell, 2009, 137(6): 1005-1017. DOI: 10.1016/j.cell.2009.04.021.
[24]	XU T, ZHU Y, XIONG Y, et al. MicroRNA-195 suppresses tumorigenicity and regulates G1/S transition of human hepatocellular carcinoma cells[J]. Hepatology, 2009, 50(1): 113-121. DOI: 10.1002/hep.22919.
[25]	XIAO F, ZHANG W, CHEN L, et al. MicroRNA-503 inhibits the G1/S transition by downregulating cyclin D3 and E2F3 in hepatocellular carcinoma[J]. J Transl Med, 2013, 11: 195. DOI: 10.1186/1479-5876-11-195.
[26]	FORNARI F, GRAMANTIERI L, FERRACIN M, et al. MiR-221 controls CDKN1C/p57 and CDKN1B/p27 expression in human hepatocellular carcinoma[J]. Oncogene, 2008, 27(43): 5651-5661. DOI: 10.1038/onc.2008.178.
[27]	LIU RF, XU X, HUANG J, et al. Down-regulation of miR-517a and miR-517c promotes proliferation of hepatocellular carcinoma cells via targeting Pyk2[J]. Cancer Lett, 2013, 329(2): 164-173. DOI: 10.1016/j.canlet.2012.10.027.
[28]	UEKI T, FUJIMOTO J, SUZUKI T, et al. Expression of hepatocyte growth factor and its receptor, the c-met proto-oncogene, in hepatocellular carcinoma[J]. Hepatology, 1997, 25(3): 619-623. DOI: 10.1002/hep.510250321.
[29]	GIORDANO S, COLUMBANO A. Met as a therapeutic target in HCC: facts and hopes[J]. J Hepatol, 2014, 60(2): 442-452. DOI: 10.1016/j.jhep.2013.09.009.
[30]	WANG Y, TAI Q, ZHANG J, et al. MiRNA-206 inhibits hepatocellular carcinoma cell proliferation and migration but promotes apoptosis by modulating cMET expression[J]. Acta Biochim Biophys Sin (Shanghai), 2019, 51(3): 243-253. DOI: 10.1093/abbs/gmy119.
[31]	LIU Y, TAN J, OU S, et al. MicroRNA-101-3p suppresses proliferation and migration in hepatocellular carcinoma by targeting the HGF/c-Met pathway[J]. Invest New Drugs, 2020, 38(1): 60-69. DOI: 10.1007/s10637-019-00766-8.
[32]	XU X, JIANG W, HAN P, et al. MicroRNA-128-3p Mediates Lenvatinib Resistance of Hepatocellular Carcinoma Cells by Downregulating c-Met[J]. J Hepatocell Carcinoma, 2022, 9: 113-126. DOI: 10.2147/JHC.S349369.
[33]	HUYNH H, ONG R, SOO KC. Foretinib demonstrates anti-tumor activity and improves overall survival in preclinical models of hepatocellular carcinoma[J]. Angiogenesis, 2012, 15(1): 59-70. DOI: 10.1007/s10456-011-9243-z.

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Expression of miR-409-3p in hepatoma HepG2 cells and its mechanism in cell proliferation

DOI: 10.3969/j.issn.1001-5256.2023.08.019