The Effect And Molecular Mechanism Of MiRNA-10a On Biological Function Of Hepatic Stellate Cells LX-2

Objective:Hepatic fibrosis is the inevitable process and pathological basis for the development of chronic liver disease into cirrhosis or even liver cancer.The proliferation and activation of hepatic stellate cells have been confirmed to be the core events in the process of liver fiber development.MiRNA-10a is a small RNA with regulation that can affect the progression of liver fibrosis in some ways.In this paper,we intend to study that the miRNA-10a regulated the activation of LX-2 cells at the cellular level;to investigate the effect of miRNA-10a on hepatic stellate cells proliferation and apoptosis;to discuss the molecular mechanism of miRNA-10a on biological function of hepatic stellate cells LX-2.The role of miRNA-10a in regulating the molecular mechanism of LX-2 cells might provide a new idea for the prevention and treatment of liver fibrosis.Methods:Human hepatic stellate cells LX-2 were used for this study.Total RNA was extracted from the cells without giving nothing or TGF-?1 stimulation for 24 h.Then the expression of miRNA-10a was detected by RT-q PCR.The miRNA-10a mimics and inhibitors were respectively transfected into LX-2,which was not stimulated or TGF-?1-stimulated.Collecting the total protein and the total RNA,the relative expression of a-SMA and Collagen I mRNA were detected by RT-qPCR.The proliferation of cells at different time after transfection was detected by CCK-8 kit.The apoptosis of cells was detected by flow cytometry after transfection 48h.The relative expression of Collagen I,a-SMA,?-catenin,GSK-3?,and p-GSK-3? protein were detected by Western blot.Result:The expression of miRNA-10a was significantly increased in the TGF-?1-stimulated LX-2 cells.RT-qPCR and Western blot results showed that miRNA-10a can promote the mRNA of Collagen I and a-SMA,and the mRNA levels or protein expressions of Collagen I,a-SMA were decreased by giving miRNA-10a inhibitor to TGF-?1-stimulated LX-2 cells.CCK-8 results showed that miRNA-10a enhanced the proliferation of LX-2 and the lower-expression of miRNA-10a inhibited the proliferation of TGF-?1-stimulated LX-2 cells.Flow cytometry results showed that miRNA-10a inhibited LX-2 apoptosis,while inhibition of miRNA-10a significantly increased TGF-?1-stimulated LX-2 apoptosis.Western blot results showed that the protein expressions of?-catenin and p-GSK-3? were increased by transfecting miRNA-10a mimics,and the protein expressions of ?-catenin and p-GSK-3? were up-regulated after giving miRNA-1 Oa inhibitors to TGF-?1-stimulated LX-2 cells.Conclusion:MiRNA-10a can promote the proliferation of hepatic stellate cells and inhibit its apoptosis.Moreover,miRNA-10a can accelerate the process of liver fibrosis by activating Wnt/?-catenin signal pathway,and it can prevent the progression of liver fibrosis when we effectively decreased the expression of miRNA-10a.This study provides a new idea for the prevention and treatment to liver fibrosis.