Functional Analysis Of MIR-101 Involved In The Pathogenesis Of Hepatocellular Carcinoma

OBJECTIVE:The aim of the study was to investigate the function and role of miR-101 in the pathogenetic process of hepatocellular carcinoma.METHODS:The gain-of-function assays were conducted through transfection with miR-101 mimics and miRNA-NC in the HepG2 and SMMC-7721 liver carcinoma cell line.The effect of miR-101 on the growth and proliferation of HepG2 and SMMC-7721 cells was assessed by Cell Counting Kit 8.Flow cytometry was performed to analyze cell cycle and apoptosis.To test the effects of miR-101 in vivo, HepG2 and SMMC-7721 cells were subcutaneously implanted into nude mice.RESULTS:HepG2 and SMMC-7721 cells were transfected with miR-101-mimics and miRNA-NC,cell proliferation assay was measured at 24,48 and 72h using the Cell Counting Kit-8,spectrometric absorbance at 490 nm was measured,miR-101 significantly inhibited proliferation of cell and the same of cell proliferation was also determined using a EdU incorporation assay, miR-101 also significantly inhibited proliferation of cell (P<0.05).The results of FCM indicated that over expresse of miR-101 inducing apoptosis of HCC cells,and miR-101 are likely to inhibit cell growth mainly by suppressing cell cycle progression, especially in the G0/G1 (P<0.05).Colony formation assay showed that miR-101 significantly inhibited the cell colony formation (P< 0.05).The migration ability of HepG2 and SMMC-7721 of scratch test and Transwell assay,miR-101 significantly inhibited cell migration (P<0.05).In animal experiments, the tumor average weight at the end of the experiment in miR-101-treated tumors was much smaller than control tumors (P< 0.05).CONCLUSION:MiR-101 overexpression in liver cancer cells decreased cell proliferation, clonogenicity, migration/invasion, and also induced G1 arrest and apoptosis. MiR-101 also inhibited tumor growth of hepatoma xenografts in nude mice. MiR-101 can be used as a tumor suppressor gene to repressing development of tumour.