In vitro anti-metastatic role of miR-10b through modulation of cytoskeleton in human hepatocellular carcinoma cell / 中华肝胆外科杂志

ABSTRACT

Objective To investigate the effect of miR-10b on migration and invasion of human hepatocellular carcinoma (HCC) cell lines.Methods Transwell assay was used to evaluate the motility and invasiveness in different HCC cell lines,qRT-PCR was then used to detect the expression levels of miR-10b in different HCC cell lines.Artificial mimics of miR-10b were transiently transfected into HepG2 cells,which have low expression of miR-10b,and then the changes in migration and invasion were evaluated by Transwell assay.Antagomirs of miR-10b (miR-10b-AS) were transiently transfected into MHCC97H cells,which have high expression of miR-10b,and then the changes in migration and invasion were evaluated as well.Cell morphology changes were detected by immunofluorescence and electron microscopy,Results There was a significant correlation of miR-10b expression level with cell motility and invasiveness.Low-level expression of miR-10b was observed in HepG2 cells,which exhibited weak motility and invasiveness; whereas high-level expression of miR-10b was observed in MHCC97H cells,which exhibited strong motility and invasiveness.Up-regulation of miR-10b expression in HepG2 cells increased cell motility and invasiveness,whereas inhibition of miR-10b reduced cell motility and invasiveness in MHCC97H cells.Immunofluorescence and electron microscopy results showed that up-regulation of miR-10b in HepG2 cells significantly increased proliferation of filopodia and lamellipodia,whereas inhibition of miR-10b decreased filopodia and lamellipodia amount in MHCC97H cells.Conclusion miR-10b is involved in the invasion and metastasis of HCC cell through regulation of cytoskeleton in vitro and inhibition of miR-10b is likely to be a new molecular target to block metastasis.