P27RF-Rho基因沉默对肝癌Bel7402细胞增殖的影响

doi:10.13481/j.1671-587x.20150321

Abstract

Abstract:

Objective To construct the RNAi plasmid targeting P27RF-Rho gene, and to explore the effect of P27RF-Rho gene silencing on the proliferation of hepatocelluar carcinoma cells Bel7402. Methods The synthetic oligonucleotide fragment was cloned into RNAi plasmid, and the constructed recombinant plasmid was transfected into the hepatocelluar carcinoma Bel7402 cells for P27RF-Rho gene silencing.This experiment was divided into Bel7402 group, Scramble-siRNA group and P27RF-Rho-siRNA group.The transfection of Bel7402 cells was observed by fluorescence microscope.The gene silencing effect was detected by RT-PCR, and the proliferation of transfected hepatocelluar carcinoma Bel7402 cells was determined by drawing growth curves, and the ability of clone formation was measured by Colony formation assay.The expression levels of P16, CyclinE, MMP-9 and CDK-5 mRNA were detected by RT-PCR. Results The expression level of P27RF-Rho gene in the Bel7402 cells in P27RF-Rho-siRNA group was obviously lower than those in Bel7402 group and Scramble-siRNA group(P<0.01).The growth speed of Bel7402 cells in P27RF-Rho-siRNA group was lower than those in Bel7402 group and Scramble-siRNA group (P<0.05).Compared with Bel7402 group and Scramble-siRNA group, the expression levels of CyclinE, MMP-9 and CDK-5 mRNA in P27RF-Rho-siRNA group were notably decreased (P<0.01), whereas the P16 mRNA expression level was significantly increased (P<0.01). Conclusion The proliferation of hepatocellular carcinoma Bel7402 cells could be controlled by inhibiting the expression of P27RF-Rho gene.

Key words: gene silencing, cell proliferation, P27RF-Rho, liver neoplasms

CLC Number:

R735.7

XING Guangyuan, XIE Shuli, QIU Wei, WANG Guangyi, LYU Guoyue, HAO Enyuan, LI Dingyang. Influence of P27RF-Rho gene silencing in proliferation of hepatocelluar carcinoma Bel7402 cells[J].Journal of Jilin University Medicine Edition, 2015, 41(03): 542-547.

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Influence of P27RF-Rho gene silencing in proliferation of hepatocelluar carcinoma Bel7402 cells

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