Effect Of RNA Interference Targeting Survivin Gene On The Proliferation Of Hepatocellular Carcinoma Cell Line HepG2

Objective:To observe the effect of the RNA interference targeting survivin gene expression on proliferation in hepatocellular carcinoma cell HepG2 and to investigate the feasibility of treatment on hepatoma with RNA interference targeting survivin gene.Methods:Three different survivin gene sequence specific shRNA vectors pshRNA-survivin -323/387/52 were constructed.Transfection of shRNA vectors was performed using the Lipofectmine2000 liposome.Its effects on the cells were evaluated by detecting following indexes:(1) The expression of survivin mRNA was detected by Reverse transcription-polymerase chain reaction(RT-PCR).The specificity of RNA interference was analyzed;(2) The vector which could significantly down-regulate the expression of survivin was transfected into HepG2 cells by Lipofectmine2000 liposome.RT-PCR was employed to detect the expression of survivin gene 24 h,48 h and 72 h after transfection.The time effect of RNA interference was analyzed;(3) The proliferation of HepG2 cells was assessed by MTT assay.Results:(1) RT-PCR showed that the pshRNA-survivin-387 vector targeting survivin gene can significantly down-regulated the expression of its mRNA in HepG2 cells when comparing with that of the controls(P＜0.05).The other vectors targeting survivin gene had no influence on survivin gene expression(P＞0.05).(2) RT-PCR showed that the expression of survivin mRNA began to decrease 24 h after transfection;and the most apparent interfering efficiency was at 48 h after transfection;at 72 h after transfection,the expression restored.(3) MTT assay showed the proliferation of HepG2 cells was inhibited.Conclusion:The pshRNA-survivin-387 vector targeting survivin gene could specifically suppress its expression and inhibit the proliferation of HepG2 cells.