Influence Of P27RF-Rho Gene Silencing In Proliferation Of Hepatocefluar Carcinoma Cells Bel7402

Objective: To construct the RNAi plasmid targeting P27RF-Rho gene, andto explore the effect of P27RF-Rho gene silencing on the proliferation ofhepatocelluar carcinoma cells Bel7402. The methods are showed as following:Methods: This experiment was divided into Bel7402group,Scramble-siRNA group(U6-shRNA-CMV-copGFP-PGK-Puro-Scramble) andP27RF-Rho-siRNA group (U6-shRNA-CMV-copGFP-PGK-Puro-P27RF-Rho).The synthetic oligonucleotide fragment was cloned into RNAi plasmid, and theconstructed recombinant plasmid was transfected into the hepatocelluarcarcinoma cells Bel7402for P27RF-Rho gene silencing. The transfection ofBel7402cells was observed by fluorescence microscope. The gene silencingeffect was detected by RT-PCR, and the proliferation of transfectedhepatocelluar carcinoma cells Bel7402was determined by drawing growthcurves，and the ability of clone formation was measured by Colony formationassay. The expression levels of P16, Cycling E, MMP9and CDK5mRNA weredetected by RT-PCR.Results: The expression level of P27RF-Rho gene in the Bel7402cells inP27RF-Rho-siRNA group was obviously lower than those in Bel7402group and Scramble-siRNA group(P＜0.01). The growth speed of Bel7402cells inP27RF-Rho-siRNA group was lower than those in Bel7402group andScramble-siRNA group (P<0.05).Compare with Bel7402group andScramble-siRNA group, the expression levels of Cyclin E, MMP9and CDK5inP27RF-Rho-siRNA group were notably decreased (P＜0.01), whereas the P16gene expression levels was significantly increased (P＜0.01).Conclusion: The proliferation of hepatocellular carcinoma cells Bel7402could be controlled by inhibiting the expression of P27RF-Rho gene.