Influence of lentiviral-mediated P27RF-Rho gene silence in invasion of liver cancer cells / 吉林大学学报(医学版)

ABSTRACT

Objective:To investigate the silencing of P27RF-Rho gene with lenvirus targeting mediated technique,and to clarify its influence in the invasion of liver cancer cells.Methods:The P27RF-Rho RNAi lentivirus was constructed. The liver cancer BEL7402 cells were infected with lentivirus. The experiment was divided into P27RF Rho-siRNA group, scramble-siRNA group and BEL7402 group.The effect of silencing P27RF-Rho gene and the expression levels of hepatocellular carcinoma (HCC)associated proteins RhoA,RhoC, VEGF,P53 and PTEN were detected;the activities of matrix metalloproteinase (MMPs)associated with tumor invasion were analyzed by Gelatin zymography;the variation of transfer ability and invasion abilities were compared by Wound healing assay experiment and Transwell experiment.Results:The Western blotting results showed the expression levels of P27RF-Rho,RhoA,RhoC,and VEGF proteins in the BEL7402 cells in experiment group were significantly lower than those in two control groups (P<0.05),and the expression levels of P53 and PTEN were higher than those in two control groups (P<0.05).The results of Gelatin zymography showed the activities of MMPs in experiment group were significantly lower than those in two control groups (P<0.01 );Wound healing assay showed that the migration ability of the BEL7402 cells in experiment group was significantly inhibited (P<0.01);the number of cells passed through the Transwell Chambers in experiment group was significantly less than those in two control groups (P<0.01).Conclusion:Silenceing P27RF-Rho can weaken the invasion ability and migration ability of human HCC BEL7402 cells.