Experimental Study On Invasive Of Hepatocellular Carcinoma Cells After Silencing P27RF-Rho Gene By Lentivirus

Objective :Silencing P27RF-Rho gene with lenvirus targeting mediated technique,and clarifying its effect on invasion of liver cancer cell.Methods:Constructing P27RF-Rho RNAi lentivirus.Infecting Liver cancer cells BEL7402 with lentivirus.The experiment is divided into three groups: BEL7402 blank group、Scramble-si RNA negative control group、P27RF Rho-si RNA experiment group.Detecting the effect of silencing P27RF-Rho gene and the expression level of hepatocellular carcinoma associated proteins Rho A、Rho C、VEGF、P53、PTEN by Western Blot;Analysising the activity of matrix metalloproteinase MMPs associated with tumor invasion by gelatin zymography;Comparing the variation of transfer ability and invasion ability by Wound healing assay experiment and Transwell experiment.Results:Western Blot results showed that the expression level of P27RF-Rho protein in the experiment group is significantly lower than that of two other control groups(P<0.05),the expression level of Rho A,Rho C,VEGF significantly are lower than that of two control groups(P<0.05),the expression level of P53 and PTEN are higher than that of two control groups(P<0.05);the results ofGelatin zymography show the activity of MMPs in experiment group is significantly lower than that of other two groups(P<0.01);Wound healing assay showed that the migration ability of experiment group cells is significantly inhibited(P < 0.01);The number of experiment group cells that pass through the Transwell Chambers are significantly less than that of other two groups,invasive ability is inhibited(P < 0.01)。Conclusion:Silenceing P27RF-Rho can weaken the transfer ability and invasion ability of human hepatoma BEL 7402 cell.