MicroRNA‑122 inhibits epithelial‑mesenchymal transition of hepatic stellate cells induced by the TGF‑β1/Smad signaling pathway

Cheng,Bianqiao; Zhu,Qi; Lin,Weiguo; Wang,Lihui

doi:10.3892/etm.2018.6962

MicroRNA‑122 inhibits epithelial‑mesenchymal transition of hepatic stellate cells induced by the TGF‑β1/Smad signaling pathway

Authors:
- Bianqiao Cheng
- Qi Zhu
- Weiguo Lin
- Lihui Wang
View Affiliations

Affiliations: Department of Hepatology, The Second Hospital of Fuzhou Affiliated Xiamen University, Fuzhou, Fujian 350007, P.R. China
Published online on: November 13, 2018 https://doi.org/10.3892/etm.2018.6962
Pages: 284-290
Copyright: © Cheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Transforming growth factor (TGF)‑β1 may stimulate the activation of hepatic stellate cells (HSCs), resulting in the development of liver fibrosis. As micro RNA (miRNA)‑122 is known to be associated with liver inflammation, its effects on the epithelial‑mesenchymal transition (EMT) of HSCs through the inhibition of the TGF‑β1/drosophila mothers against decapentaplegic protein 4 (Smad4) signaling pathway were investigated. The MTT assay was performed to explore the optimum TGF‑β1 concentration suitable for HSC stimulation. Fluorescence microscopy was used to observe the transfection efficiency and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis were used to observe gene and protein expression levels of α‑smooth muscle actin (α‑SMA), E‑cadherin, N‑cadherin and Smad4, respectively, in HSCs treated with TGF‑β1 or TGF‑β1 and miRNA‑122. MTT assay results indicated that the concentration of 10 µg/l TGF‑β1 was suitable for maximum growth and survival of HSCs. Notably, the mRNA expression levels of N‑cadherin and α‑SMA were significantly increased (each, P<0.05), but the expression levels of E‑cadherin were decreased following 10 µg/l TGF‑β1 treatment. Similar results were observed regarding the protein expression levels of N‑cadherin, α‑SMA and E‑cadherin. Furthermore, the expression of F‑actin was increased in the 10 µg/l TGF‑β1 treated group compared with the 0 µg/l TGF‑β1 treaded group and stretching of the muscle fiber filament was observed. miRNA‑122 lentiviral vector transfection significantly decreased the mRNA expression of N‑cadherin and increased the mRNA expression of E‑cadherin in HSCs stimulated with TGF‑β1, as evident from RT‑qPCR results. Similar results were also observed regarding the protein expression levels of N‑cadherin and E‑cadherin. The expression levels of Smad4, the primary component of the TGF‑β1 signaling pathway, were significantly lower in cells treated with TGF‑β1 and miRNA‑122 (P<0.01) compared those treated with TGF‑β1. Thus, miRNA‑122 may inhibit the activation and EMT of HSCs stimulated by TGF‑β1.

View Figures

View References

1	Tomita K, Teratani T, Suzuki T, Shimizu M, Sato H, Narimatsu K, Usui S, Furuhashi H, Kimura A, Nishiyama K, et al: Acyl-CoA: Cholesterol acyltransferase 1 mediates liver fibrosis by regulating free cholesterol accumulation in hepatic stellate cells. J Hepatol. 61:98–106. 2014. View Article : Google Scholar : PubMed/NCBI
2	Duval F, Moreno-Cuevas JE, González-Garza MT, Rodríguez-Montalvo C and Cruz-Vega DE: Liver fibrosis and protection mechanisms action of medicinal plants targeting apoptosis of hepatocytes and hepatic stellate cells. Adv Pharmacol Sci. 2014:3732952014.PubMed/NCBI
3	Yang AT, Hu DD, Wang P, Cong M, Liu TH, Zhang D, Sun YM, Zhao WS, Jia JD and You H: TGF-β1 induces the dual regulation of hepatic progenitor cells with both anti- and proliver fibrosis. Stem Cells Int. 2016:14926942016. View Article : Google Scholar : PubMed/NCBI
4	Li JH, Huang XR, Zhu HJ, Johnson R and Lan HY: Role of TGF-beta signaling in extracellular matrix production under high glucose conditions. Kidney Int. 63:2010–2019. 2003. View Article : Google Scholar : PubMed/NCBI
5	Dropmann A, Dediulia T, Breitkopf-Heinlein K, Korhonen H, Janicot M, Weber SN, Thomas M, Piiper A, Bertran E, Fabregat I, et al: TGF-β1 and TGF-β2 abundance in liver diseases of mice and men. Oncotarget. 7:19499–19518. 2016. View Article : Google Scholar : PubMed/NCBI
6	Duan L, Ye L, Zhuang L, Zou X, Liu S, Zhang Y, Zhang L, Jin C and Huang Y: VEGFC/VEGFR3 axis mediates TGFβ1-induced epithelial-to-mesenchymal transition in non-small cell lung cancer cells. PLoS One. 13:e02004522018. View Article : Google Scholar : PubMed/NCBI
7	Wang QL, Tao YY, Yuan JL, Shen L and Liu CH: Salvianolic acid B preventsepithelial-to-mesenchymal transition through the TGF-beta1 signal transduction pathway in vivo and in vitro. BMC Cell Biol. 11:312010. View Article : Google Scholar : PubMed/NCBI
8	Bi WR, Yang CQ and Shi Q: Transforming growth factor-β1 induced epithelial-mesenchymal transition in hepatic fibrosis. Hepatogastroenterology. 59:1960–1963. 2012.PubMed/NCBI
9	Girard M, Jacquemin E, Munnich A, Lyonnet S and Henrion-Caude A: miR-122, a paradigm for the role of microRNAs in the liver. J Hepatol. 48:648–656. 2008. View Article : Google Scholar : PubMed/NCBI
10	Zhang Y, Jia Y, Zheng R, Guo Y, Wang Y, Guo H, Fei M and Sun S: Plasma microRNA-122 as a biomarker for viral-, alcohol-, and chemical-related hepatic diseases. Clin Chem. 56:1830–1838. 2010. View Article : Google Scholar : PubMed/NCBI
11	Lewis Starkey PJ, Dear J, Platt V, Simpson KJ, Craig DG, Antoine DJ, French NS, Dhaun N, Webb DJ, Costello EM, et al: Circulating microRNAs as potential markers of human drug-induced liver injury. Hepatology. 54:1767–1776. 2011. View Article : Google Scholar : PubMed/NCBI
12	Su TH, Chen M, Liu CJ, Chen CL, Ting TT, Tseng TC, Chen PJ, Kao JH and Chen DS: Serum microRNA-122 level correlates with Virologic responses to pegylated interferon therapy in chronic hepatitis C. Proc Natl Acad Sci USA. 110:7844–7849. 2013. View Article : Google Scholar : PubMed/NCBI
13	Ding X, Ding J, Ning J, Yi F, Chen J, Zhao D, Zheng J, Liang Z, Hu Z and Du Q: Circulating microRNA-122 as a potential biomarker for liver injury. Mol Med Rep. 5:1428–1432. 2012.PubMed/NCBI
14	Omran AA, Osman KS, Kamel HM, Abdel-Naem EA and Hasan DE: MicroRNA-122 as a novel non-invasive marker of liver fibrosis in hepatitis C virus patients. Clin Lab. 62:1329–1337. 2016. View Article : Google Scholar : PubMed/NCBI
15	Nakamura M, Kanda T, Jiang X, Haga Y, Takahashi K, Wu S, Yasui S, Nakamoto S and Yokosuka O: Serum microRNA-122 and Wisteria floribunda agglutinin-positive Mac-2 binding protein are useful tools for liquid biopsy of the patients with hepatitis B virus and advanced liver fibrosis. PLoS One. 12:e01773022017. View Article : Google Scholar : PubMed/NCBI
16	Waidmann O, Köberle V, Brunner F, Zeuzem S, Piiper A and Kronenberger B: Serum microRNA-122 predicts survival in patients with liver cirrhosis. PLoS One. 7:e456522012. View Article : Google Scholar : PubMed/NCBI
17	Tsai WC, Hsu PW, Lai TC, Chau GY, Lin CW, Chen CM, Lin CD, Liao YL, Wang JL, Chau YP, et al: MicroRNA-122, a tumor suppressor microRNA that regulates intrahepatic metastasis of hepatocellular carcinoma. Hepatology. 49:1571–1582. 2009. View Article : Google Scholar : PubMed/NCBI
18	Qiao DD, Yang J, Lei XF, Mi GL, Li SL, Li K, Xu CQ and Yang HL: Expression of microRNA-122 and microRNA-22 in HBV-related liver cancer and the correlation with clinical features. Eur Rev Med Pharmacol Sci. 21:742–747. 2017.PubMed/NCBI
19	Wang N, Wang Q, Shen D, Sun X, Cao X and Wu D: Downregulation of microRNA-122 promotes proliferation, migration, and invasion of human hepatocellular carcinoma cells by activating epithelial-mesenchymal transition. Onco Targets Ther. 9:2035–2047. 2016. View Article : Google Scholar : PubMed/NCBI
20	Xu J, Zhu X, Wu L, Yang R, Yang Z, Wang Q and Wu F: MicroRNA-122 suppresses cell proliferation and induces cell apoptosis in hepatocellular carcinoma by directly targeting Wnt/β-catenin pathway. Liver Int. 32:752–760. 2012. View Article : Google Scholar : PubMed/NCBI
21	Jin Y, Wang J, Han J, Luo D and Sun Z: MiR-122 inhibits epithelial-mesenchymal transition in hepatocellular carcinoma by targeting Snail1 and Snail2 and suppressing WNT/β-cadherin signaling pathway. Exp Cell Res. 360:210–217. 2017. View Article : Google Scholar : PubMed/NCBI
22	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
23	Ma L, Yang X, Wei R, Ye T, Zhou JK, Wen M, Wen RT, Li P, Dong B, Liu L, et al: MicroRNA-214 promotes hepatic stellate cell activation and liver fibrosis by suppressing Sufu expression. Cell Death Dis. 9:7182018. View Article : Google Scholar : PubMed/NCBI
24	Cui X, Zhang X, Yin Q, Meng A, Su S, Jing X, Li H, Guan X, Li X, Liu S and Cheng M: F-actin cytoskeleton reorganization is associated with hepatic stellate cell activation. Mol Med Rep. 9:1641–1647. 2014. View Article : Google Scholar : PubMed/NCBI
25	Yu F, Ji S, Su L, Wan L, Zhang S, Dai C, Wang Y, Fu J and Zhang Q: Adipose-derived mesenchymal stem cells inhibit activation of hepatic stellate cells in vitro and ameliorate rat liver fibrosis in vivo. J Formos Med Assoc. 114:130–138. 2015. View Article : Google Scholar : PubMed/NCBI
26	Yu H, Shen Y, Hong J, Xia Q, Zhou F and Liu X: The contribution of TGF-β in Epithelial-Mesenchymal Transition (EMT): Down-regulation of E-cadherin via snail. Neoplasma. 62:1–15. 2015. View Article : Google Scholar : PubMed/NCBI
27	Chen T, Nie HY, Gao X, Yang J, Pu J, Chen Z, Cui X, Wang Y, Wang H and Jia G: Epithelial-mesenchymal transition involved in pulmonary fibrosis induced by multi-walled carbon nanotubes via TGF-beta/Smad signaling pathway. Toxicol Lett. 226:150–162. 2014. View Article : Google Scholar : PubMed/NCBI
28	Molè-Bajer J, Bajer AS and Inoué S: Three-dimensional localization and redistribution of F-actin in higher plant mitosis and cell plate formation. Cell Motil Cytoskeleton. 10:217–228. 1988. View Article : Google Scholar : PubMed/NCBI
29	Gong X, Fan Y, Zhang Y, Luo C, Duan X, Yang L and Pan J: Inserted rest period resensitizes MC3T3-E1 cells to fluid shear stress in a time-dependent manner via F-actin-regulated mechanosensitive channel(s). Biosci Biotechnol Biochem. 78:565–573. 2014. View Article : Google Scholar : PubMed/NCBI
30	Lee UE and Friedman SL: Mechanisms of hepatic fibrogenesis. Best Pract Res Clin Gastroenterol. 25:195–206. 2011. View Article : Google Scholar : PubMed/NCBI
31	Boesch-Saadatmandi C, Wagner AE, Wolffram S and Rimbach G: Effect of quercetin on inflammatory gene expression in mice liver in vivo-role of redox factor 1, miRNA-122 and miRNA-125b. Pharmacol Res. 65:523–530. 2012. View Article : Google Scholar : PubMed/NCBI
32	Bala S, Petrasek J, Ward J, Alao H, Levin I and Szabo G: Serum microrna-122 and mir-155 as biomarkers of liver injury and inflammation in models of acute and chronic liver disease. Gastroenterology. 140:9062012. View Article : Google Scholar