siRNA对HepG2细胞survivin表达的抑制作用

目的　研究siRNA对肝癌细胞株HepG2中survivin基因表达的抑制作用。 方法　设计、合成一对survivin编码基因的反向重复序列 ,中间间隔 9个核苷酸序列 ,通过定向克隆至载体PSilencer 2 .1,构建siRNA真核表达载体 ,经稳定转染HepG2细胞后应用RT PCR及流式细胞术检测肝癌细胞中survivin基因mRNA及蛋白质表达的抑制情况。 结果　测序证实siRNA真核表达载体构建成功。通过RT PCR及流式细胞术检测 ,siRNA在mRNA及蛋白质水平抑制survivin基因表达分别达 73%和 75 %。结论　构建的PSilencer( ) survivin重组质粒能有效抑制survivin基因在肝癌细胞株HepG2中的表达 ,从而为肿瘤的生物学治疗提供了新的方法和材料。

Inhibition effect of siRNA on survivin expression in HepG2 cells

Objective To study the inhibition effect of siRNA on the survivin expression in HepG2 cells. Methods One pair of 21bp reverse repeated motifs of survivin target sequence with 9 spacer were synthesized and inserted into plasmid Psilencer2.1 to generate siRNA eukaryotic expression vector. After stable transfection into HepG2 cells , the survivin mRNA and protein expression inhibition was detected by use of RT-PCR and flow cytometer. Results The recombinant plasmid P Silencer( )-survivin was successfully constructed. By use of RT-PCR and flow cytometer detection , survivin mRNA and protein expression inhibition ratio reached 73% and 75% respectively. Conclusion siRNA targeting survivin gene can specially suppress its expression in HepG2 cells. This provides a new method and material to the biological therapy of cancer.