miR-93-5p promotes insulin resistance to regulate type 2 diabetes progression in HepG2 cells by targeting HGF

Zhou,Man; Hou,Yilin; Wu,Jun; Li,Guangli; Cao,Ping; Chen,Wan; Hu,Lingli; Gan,Dingyun

doi:10.3892/mmr.2021.11968

miR-93-5p promotes insulin resistance to regulate type 2 diabetes progression in HepG2 cells by targeting HGF

Authors:
- Man Zhou
- Yilin Hou
- Jun Wu
- Guangli Li
- Ping Cao
- Wan Chen
- Lingli Hu
- Dingyun Gan
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Affiliations: Department of Endocrinology, Wuhan Third Hospital, Wuhan, Hubei 430060, P.R. China
Published online on: March 8, 2021 https://doi.org/10.3892/mmr.2021.11968
Article Number: 329
Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Insulin resistance is a common feature of type 2 diabetes mellitus (T2DM). However, the mechanisms underlying insulin resistance are not completely understood. The present study aimed to investigate the effect of microRNA (miR)‑93‑5p on insulin resistance in T2DM cells. Human hepatocellular carcinoma (HCC; HepG2) cells were cultured in medium with high glucose content (30 mM glucose) to establish an in vitro insulin‑resistant cell model (IR group). Glucose consumption and glycogen synthesis assays were performed to assess glucose consumption and glycogen synthesis, respectively. By performing immunoprecipitation assays, the abundance of the Met‑insulin receptor complex was detected in HepG2 cells. miR‑93‑5p and hepatocyte growth factor (HGF) mRNA expression levels were measured via reverse transcription‑quantitative PCR, and HGF protein expression levels were measured via western blotting. A dual‑luciferase reporter assay was conducted to investigate the interaction between miR‑93‑5p and HGF. Cell Counting Kit‑8, BrdU and caspase‑3 activity assays were performed to evaluate cell viability, proliferation and apoptosis, respectively, in insulin‑resistant HepG2 cells following transfection with small interfering RNA‑HGF, HGF overexpression vector, miR‑93‑5p mimic or miR‑93‑5p inhibitor. The results demonstrated that miR‑93‑5p expression was significantly increased and HGF expression was significantly decreased in HCC tissues isolated from patients with or without T2DM compared with adjacent healthy tissues isolated from patients without T2DM. Compared with the IR group, miR‑93‑5p overexpression significantly increased cell proliferation, glucose consumption and glycogen synthesis, but significantly inhibited apoptosis in insulin‑resistant HepG2 cells. By contrast, compared with the IR group, HGF overexpression significantly inhibited cell proliferation, glucose consumption and glycogen synthesis, but significantly enhanced cell apoptosis in insulin‑resistant HepG2 cells. Following co‑transfection with HGF overexpression vector and miR‑93‑5p mimic, miR‑93‑5p mimic‑mediated induction of HepG2 cell proliferation, glucose consumption and glycogen synthesis in insulin‑resistant HepG2 cells was inhibited. Collectively, the results of the present study indicated that miR‑93‑5p enhanced insulin resistance to regulate T2DM progression in HepG2 cells by targeting HGF.

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1	Triplitt CL: Examining the mechanisms of glucose regulation. Am J Manag Care. 18 (Suppl 1):S4–S10. 2012.PubMed/NCBI
2	Chatterjee S, Khunti K and Davies MJ: Type 2 diabetes. Lancet. 389:2239–2251. 2017. View Article : Google Scholar : PubMed/NCBI
3	Petersen MC and Shulman GI: Mechanisms of insulin action and insulin resistance. Physiol Rev. 98:2133–2223. 2018. View Article : Google Scholar : PubMed/NCBI
4	Chan JC, Malik V, Jia W, Kadowaki T, Yajnik CS, Yoon KH and Hu FB: Diabetes in Asia: Epidemiology, risk factors, and pathophysiology. JAMA. 301:2129–2140. 2009. View Article : Google Scholar : PubMed/NCBI
5	Riddy DM, Delerive P, Summers RJ, Sexton PM and Langmead CJ: G Protein-coupled receptors targeting insulin resistance, obesity, and type 2 diabetes mellitus. Pharmacol Rev. 70:39–67. 2018. View Article : Google Scholar : PubMed/NCBI
6	Li G, Zhou F, Chen Y, Zhang W and Wang N: Kukoamine A attenuates insulin resistance and fatty liver through downregulation of Srebp-1c. Biomed Pharmacother. 89:536–543. 2017. View Article : Google Scholar : PubMed/NCBI
7	von Horn C and Minor T: Modern concepts for the dynamic preservation of the liver and kidneys in the context of transplantation. Pathologe. 40:292–298. 2019.(In German). View Article : Google Scholar : PubMed/NCBI
8	El-Serag HB, Hampel H and Javadi F: The association between diabetes and hepatocellular carcinoma: a systematic review of epidemiologic evidence. Clin Gastroenterol Hepatol. 4:369–380. 2006. View Article : Google Scholar : PubMed/NCBI
9	Polesel J, Zucchetto A, Montella M, Dal Maso L, Crispo A, La Vecchia C, Serraino D, Franceschi S and Talamini R: The impact of obesity and diabetes mellitus on the risk of hepatocellular carcinoma. Ann Oncol. 20:353–357. 2009. View Article : Google Scholar : PubMed/NCBI
10	Ueki T, Kaneda Y, Tsutsui H, Nakanishi K, Sawa Y, Morishita R, Matsumoto K, Nakamura T, Takahashi H, Okamoto E, et al: Hepatocyte growth factor gene therapy of liver cirrhosis in rats. Nat Med. 5:226–230. 1999. View Article : Google Scholar : PubMed/NCBI
11	Fafalios A, Ma J, Tan X, Stoops J, Luo J, Defrances MC and Zarnegar R: A hepatocyte growth factor receptor (Met)-insulin receptor hybrid governs hepatic glucose metabolism. Nat Med. 17:1577–1584. 2011. View Article : Google Scholar : PubMed/NCBI
12	Zhang YY, Li C, Yao GF, Du LJ, Liu Y, Zheng XJ, Yan S, Sun JY, Liu Y, Liu MZ, et al: Deletion of Macrophage Mineralocorticoid Receptor Protects Hepatic Steatosis and Insulin Resistance Through ERα/HGF/Met Pathway. Diabetes. 66:1535–1547. 2017. View Article : Google Scholar : PubMed/NCBI
13	Oliveira AG, Araújo TG, Carvalho BM, Rocha GZ, Santos A and Saad MJ: The role of hepatocyte growth factor (HGF) in insulin resistance and diabetes. Front Endocrinol (Lausanne). 9:5032018. View Article : Google Scholar : PubMed/NCBI
14	Li Z, Xu R and Li N: MicroRNAs from plants to animals, do they define a new messenger for communication? Nutr Metab (Lond). 15:682018. View Article : Google Scholar : PubMed/NCBI
15	Reinhart BJ, Weinstein EG, Rhoades MW, Bartel B and Bartel DP: MicroRNAs in plants. Genes Dev. 16:1616–1626. 2002. View Article : Google Scholar : PubMed/NCBI
16	Nicolai S, Pieraccioli M, Smirnov A, Pitolli C, Anemona L, Mauriello A, Candi E, Annicchiarico-Petruzzelli M, Shi Y, Wang Y, et al: ZNF281/Zfp281 is a target of miR-1 and counteracts muscle differentiation. Mol Oncol. 14:294–308. 2020. View Article : Google Scholar : PubMed/NCBI
17	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
18	Fischer C, Seki T, Lim S, Nakamura M, Andersson P, Yang Y, Honek J, Wang Y, Gao Y, Chen F, et al: A miR-327-FGF10-FGFR2-mediated autocrine signaling mechanism controls white fat browning. Nat Commun. 8:20792017. View Article : Google Scholar : PubMed/NCBI
19	Trajkovski M, Hausser J, Soutschek J, Bhat B, Akin A, Zavolan M, Heim MH and Stoffel M: MicroRNAs 103 and 107 regulate insulin sensitivity. Nature. 474:649–653. 2011. View Article : Google Scholar : PubMed/NCBI
20	Pardo F, Villalobos-Labra R, Sobrevia B, Toledo F and Sobrevia L: Extracellular vesicles in obesity and diabetes mellitus. Mol Aspects Med. 60:81–91. 2018. View Article : Google Scholar : PubMed/NCBI
21	Thurnherr T, Mah WC, Lei Z, Jin Y, Rozen SG and Lee CG: Differentially expressed miRNAs in hepatocellular carcinoma target genes in the genetic information processing and metabolism pathways. Sci Rep. 6:200652016. View Article : Google Scholar : PubMed/NCBI
22	Wang X, Liao Z, Bai Z, He Y, Duan J and Wei L: MiR-93-5p Promotes cell proliferation through down-regulating PPARGC1A in hepatocellular carcinoma cells by bioinformatics analysis and experimental verification. Genes (Basel). 9:92018. View Article : Google Scholar
23	Aravinthan A, Challis B, Shannon N, Hoare M, Heaney J and Alexander GJ: Selective insulin resistance in hepatocyte senescence. Exp Cell Res. 331:38–45. 2015. View Article : Google Scholar : PubMed/NCBI
24	Williams JF and Olefsky JM: Defective insulin receptor function in down-regulated HepG2 cells. Endocrinology. 127:1706–1717. 1990. View Article : Google Scholar : PubMed/NCBI
25	Wang H, Huo X, Yang XR, He J, Cheng L, Wang N, Deng X, Jin H, Wang N, Wang C, et al: STAT3-mediated upregulation of lncRNA HOXD-AS1 as a ceRNA facilitates liver cancer metastasis by regulating SOX4. Mol Cancer. 16:1362017. View Article : Google Scholar : PubMed/NCBI
26	Darpolor MM, Basu SS, Worth A, Nelson DS, Clarke-Katzenberg RH, Glickson JD, Kaplan DE and Blair IA: The aspartate metabolism pathway is differentiable in human hepatocellular carcinoma: Transcriptomics and (13) C-isotope based metabolomics. NMR Biomed. 27:381–389. 2014. View Article : Google Scholar : PubMed/NCBI
27	Schmidt JA, Avarbock MR, Tobias JW and Brinster RL: Identification of glial cell line-derived neurotrophic factor-regulated genes important for spermatogonial stem cell self-renewal in the rat. Biol Reprod. 81:56–66. 2009. View Article : Google Scholar : PubMed/NCBI
28	Zhu HR, Yu XN, Zhang GC, Shi X, Bilegsaikhan E, Guo HY, Liu LL, Cai Y, Song GQ, Liu TT, et al: Comprehensive analysis of long non coding RNA messenger RNA microRNA co expression network identifies cell cycle related lncRNA in hepatocellular carcinoma. Int J Mol Med. 44:1844–1854. 2019.PubMed/NCBI
29	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
30	Tsai S, Clemente-Casares X, Zhou AC, Lei H, Ahn JJ, Chan YT, Choi O, Luck H, Woo M, Dunn SE, et al: Insulin Receptor-Mediated Stimulation Boosts T Cell Immunity during Inflammation and Infection. Cell Metab. 28:922–934.e4. 2018. View Article : Google Scholar : PubMed/NCBI
31	Mantovani A and Targher G: Type 2 diabetes mellitus and risk of hepatocellular carcinoma: Spotlight on nonalcoholic fatty liver disease. Ann Transl Med. 5:2702017. View Article : Google Scholar : PubMed/NCBI
32	Sim WJ, Iyengar PV, Lama D, Lui SKL, Ng HC, Haviv-Shapira L, Domany E, Kappei D, Tan TZ, Saei A, et al: c-Met activation leads to the establishment of a TGFβ-receptor regulatory network in bladder cancer progression. Nat Commun. 10:43492019. View Article : Google Scholar : PubMed/NCBI
33	Hartmann S, Bhola NE and Grandis JR: HGF/Met Signaling in head and neck cancer: Impact on the tumor microenvironment. Clin Cancer Res. 22:4005–4013. 2016. View Article : Google Scholar : PubMed/NCBI
34	Boromand N, Hasanzadeh M, ShahidSales S, Farazestanian M, Gharib M, Fiuji H, Behboodi N, Ghobadi N, Hassanian SM, Ferns GA, et al: Clinical and prognostic value of the C-Met/HGF signaling pathway in cervical cancer. J Cell Physiol. 233:4490–4496. 2018. View Article : Google Scholar : PubMed/NCBI
35	Coudriet GM, Stoops J, Orr AV, Bhushan B, Koral K, Lee S, Previte DM, Dong HH, Michalopoulos GK, Mars WM, et al: A noncanonical role for plasminogen activator inhibitor type 1 in obesity-induced diabetes. Am J Pathol. 189:1413–1422. 2019. View Article : Google Scholar : PubMed/NCBI
36	Muratsu J, Iwabayashi M, Sanada F, Taniyama Y, Otsu R, Rakugi H and Morishita R: Hepatocyte growth factor prevented high-fat diet-induced obesity and improved insulin resistance in mice. Sci Rep. 7:1302017. View Article : Google Scholar : PubMed/NCBI
37	Bertola A, Bonnafous S, Cormont M, Anty R, Tanti JF, Tran A, Le Marchand -Brustel Y and Gual P: Hepatocyte growth factor induces glucose uptake in 3T3-L1 adipocytes through A Gab1/phosphatidylinositol 3-kinase/Glut4 pathway. J Biol Chem. 282:10325–10332. 2007. View Article : Google Scholar : PubMed/NCBI
38	Calin GA and Croce CM: MicroRNA signatures in human cancers. Nat Rev Cancer. 6:857–866. 2006. View Article : Google Scholar : PubMed/NCBI
39	Ma DH, Li BS, Liu JJ, Xiao YF, Yong X, Wang SM, Wu YY, Zhu HB, Wang DX and Yang SM: miR-93-5p/IFNAR1 axis promotes gastric cancer metastasis through activating the STAT3 signaling pathway. Cancer Lett. 408:23–32. 2017. View Article : Google Scholar : PubMed/NCBI
40	Huang W, Yang Y, Wu J, Niu Y, Yao Y, Zhang J, Huang X, Liang S, Chen R, Chen S, et al: Circular RNA cESRP1 sensitises small cell lung cancer cells to chemotherapy by sponging miR-93-5p to inhibit TGF-β signalling. Cell Death Differ. 27:1709–1727. 2020. View Article : Google Scholar : PubMed/NCBI
41	Zhang S, He Y, Liu C, Li G, Lu S, Jing Q, Chen X, Ma H, Zhang D, Wang Y, et al: miR-93-5p enhances migration and invasion by targeting RGMB in squamous cell carcinoma of the head and neck. J Cancer. 11:3871–3881. 2020. View Article : Google Scholar : PubMed/NCBI
42	Chen C, Zheng Q, Kang W and Yu C: Long non-coding RNA LINC00472 suppresses hepatocellular carcinoma cell proliferation, migration and invasion through miR-93-5p/PDCD4 pathway. Clin Res Hepatol Gastroenterol. 43:436–445. 2019. View Article : Google Scholar : PubMed/NCBI
43	Vauthey JN, Lauwers GY, Esnaola NF, Do KA, Belghiti J, Mirza N, Curley SA, Ellis LM, Regimbeau JM, Rashid A, et al: Simplified staging for hepatocellular carcinoma. J Clin Oncol. 20:1527–1536. 2002. View Article : Google Scholar : PubMed/NCBI