MiR-16-5p Inhibits The Proliferation,Invasion And Migration Of Liver Cancer Cell HepG2 By Targeting VEGFA

Objective:To detect the expression of mi R-16-5p in primary liver cancer tissues and its effect on proliferation,invasion and migration of HepG2 cell and to explore the related molccular mechanism.Methods:1.qRT-PCR was used to detect the expression of miR-16-5p in PLC tissues,non-tumor tissues,liver cancer cells and liver cells.2.Effect of up-regulating miR-16-5p on the proliferation,invasion and migration abilities of HepG2 cell were detected with RTCA system,Transwell assay and wound healing assay,respectively.3.Predicting miR-16-5p downstream target genes by online bioinformatics datebase and validating it by double luciferase reports and Western blot assay.4.The expression of EMT-related proteins were detected by western blot after up-rugulation of miR-16-5p.5.Cell experiments were performed to determine the effect of VEGFA on the function of miR-16-5p.Results:1.The expression of miR-16-5p in PLC cancer tissues is significantly lower than adjacent tissues(P<0.01)and it is lower in liver cancer cell lines(HepG2,HuH-7)than nomal liver cell line(HL-7702)(P<0.001).2.The proliferation,invasion and migration abilities of HepG2 cell were significantly reduced after up-regulating miR-16-5p.3.Online datebase predicts that 3'-UTR of VEGFA contains miR-16-5p binding site,and VEGFA may be a target gene of miR-16-5p.4.After co-transfection of VEGFA wildtype and miR mimics,the luciferase activity was significantly lower than control group(P<0.05).5.The VEGFA,N-cadherin and Vimentin protein expression decreased(P<0.001)and the E-cadherin protein expression increased(P<0.05)after up-regulating miR-16-5p.6.After overexpression of VEGFA,miR-16-5p's tumor suppressive effect was significantly reduced.(proliferation P<0.05,invasion P<0.05,migration P<0.01)Conclusion:1.miR-16-5p expression decreased in PLC tissues and liver cancer cell.2.Overexpression of miR-16-5p can lead to a decrease in VEGFA protein expression,and bioinformatics prediction as well as double luciferase assay indicate that VEGFA is a down stream target gene of miR-16-5p.3.miR-16-5p can be used as tumor suppressor to regulate the proliferation,incasion,,migration and EMT of HepG2 cell.This effect may be related to the miR-16-5p/VEGFA axis.