siRNA抑制survivin基因及其对人肝癌细胞株HepG2细胞凋亡的影响

siRNA inhibits survivin gene and its effect on the apoptosis of HepG2 cells

目的观察特异小干扰RNA(small interfering RNA,siRNA)抑制survivin基因表达对人肝癌细胞株HepG2细胞凋亡的影响。方法构建survivin-siRNA真核表达载体,利用脂质体转染人肝癌HepG2细胞,采用反转录聚合酶链反应(RT-PCR)技术检测survivin基因mRNA表达水平。针对转染后不同的时间点采用四甲基偶氮唑蓝(MTT)法检测各组细胞的增殖以及HepG2细胞的凋亡情况。结果构建的survivin-siRNA转染组survivin蛋白表达较对照组明显下调,差异有显著性(P<0.01),其增殖能力较对照组显著下降(P<0.001);而阳性对照组及未转染组对survivin表达及mRNA抑制作用不明显(P=0.219)。结论所构建的针对人survivin-siRNA可显著抑制survivin的表达及HepG2细胞凋亡。

Objective To study the effect of small interfering RNA（siRNA）of human survivin gene on apoptosis of human hepatoma cell line HepG2. Methods The PCR products containing siRNA sequence were transfected into HepG2 via Lipofectamine TM 2000, cell viability was determined by MTT assay. Apoptosis was detected by morphological observation and flow cytometry analysis. The expression level of HepG2 mRNA was assayed by RT-PCR. Result The cell growth and viability of siRNA transfected group were significantly inhibited when compared with that of the negative and blank control groups（P ＆lt; 0.01）, whereas the latter two groups had simi1ar expression levels. Conclusion Interference with siRNA against survivin gene can effectively inhibit the specific gene expression, which may play a significant role in inducing of apoptosis of the corresponding HepG2 cells.