Abstract:

Objective To explore the effect of microRNA-373（miR-373）on the expression of large tumor suppressor 2（LATS2）and its influence on the proliferation and apoptosis of hepatocellular carcinoma cell line HepG2. Methods The miR-373 level was detected in HepG2 cells and normal hepatocytes LO2 by fluorescence real-time quantitative PCR（QPCR）. HepG2 cells were assigned into inhibitor group and control group，and transfected with miR-373 inhibitors or negative control（NC）by Lipofectamine liposome method. The miR-373 levels of both groups were detected by QPCR in order to evaluate the transfection efficiency. MTT method and flow cytometry were used to detect the cell proliferation and apoptosis. The expressions of apoptosis related genes（Bax and caspase-3）and LATS2 in each group were detected by Western blotting. The relationship between miR-373 and LATS2 was verified using double luciferase reporter assay. Results Compared with LO2 cells，the level of miR-373 in HepG2 cells was increased，and the difference was statistically significant（P＜0.05）. The level of miR-373 in the inhibition group was lower than that of control group（P＜0.05）， indicating that the miR373 expression of HepG2 cells was inhibited successfully. Compared with control group，there were decreased proliferative rate, and increased apoptotic rate and protein levels of Bax，caspase-3 and LATS2 in inhibitor group（P＜0.05）. Dual luciferase reporter assay showed that miR-373 could significantly inhibit the luciferase activity of cells transfected with wild-type LATS2-3’UTR，and had no effect on luciferase activity of cells transfected with mutant LATS2-3’ UTR. Conclusion MiR-373 can regulate the expression of LATS2，and inhibiting the expression of miR-373 can inhibit the proliferation and induce apoptosis of HepG2 cells，providing some reference for the targeted therapy of hepatocellular carcinoma.