siRNA阻抑Survivin基因表达对肝癌细胞周期影响的研究

Research on influence of expression inhibition of Survivin gene by siRNA on liver cancer cell cycles

目的采用RNA干扰(RNAi)技术抑制人肝癌HepG2细胞中Survivin基因表达,观察小分子干扰RNA(siRNA)对肝癌细胞周期的影响。方法构建人肝癌HepG2的survivin-siRNA,将其导入肝癌细胞系后研究。HepG2细胞分为四组:siRNA干扰组、siRNA干扰对照组、未转染的对照组、空白载体组。作用48h后Western印迹检测基因沉默效果,MTT法测量细胞生长情况,流式细胞术分析siRNA对肝癌细胞增殖周期的影响。结果 Western印迹法显示干扰组survivin基因的表达受到明显抑制。MTT法显示经survivin基因干扰后的HepG2细胞相比对照组细胞吸光度A值较低;尤其是24、48小时降低更为明显(P<0.05)。流式细胞术显示干扰组的G2/M细胞百分率相对干扰对照组显着增大(P<0.05)。结论 siRNA转染能够抑制细胞survivin基因表达,使人肝癌细胞株HepG2细胞阻滞于G2/M期。

Objective To inhibit the expression of Survivn gene in hepatocellular carcinoma cell line HepG2 by RNA interference technology,to observe small interfering RNA targeting on the hepatic carcinoma cell cycle.Methods Structure Survivin-siRNA aimed at human hepatocellular carcinoma cell line HepG2.Then siRNA was introduced into hepatocellular carcinoma cell line and to study below.Divided the HepG2 cells into 4 groups：siRNA interference group,siRNA interference control group,non-transfected group,null group as comparison.After 48h,detected gene silence effects by Western blot,measured growth case of cells by MTT method and analyzed effect of siRNA on hepatocarcinoma cell generation cycle by flow cytometry.Results It showed that expression on Survivin in interference group was inhibited obviously by Western blot.The result of MTT method showed that absorbance A of HepG2 cell reduced compared to the comparison group after gene interfered by Survivin,especially at the time of 24h and 48h,this case was more obviously（P0.05）.Percentage of G2/M cells siRNA in interference group was significantly increased relative to it in siRNA interference control group,which showed by flow cytometry（P0.05）.Conclusions Conclusion： siRNA transfixion could inhibit the expression of Survivin gene,which leads HepG2 cells are blocked in stage G2/M.