Survivin siRNA对人肺腺癌细胞SPCA1和SH77抑制作用的比较分析

Inhibitory Effects of the Survivin siRNA Transfection on Human Lung Adenocarcinoma Cells SPCA1 and SH77

背景与目的 Survivin是IAP家族的新成员,具有抑制凋亡和调节细胞增殖的双重作用,是迄今发现最强的凋亡抑制因子。本项研究的目的是通过将survivin的小干扰RNA(small interfering RNA,siRNA)表达载体分别转染至肺癌细胞SPCA1和SH77,分析survivin siRNA对不同肺癌细胞的增殖抑制作用。方法构建靶向survivin的siRNA表达载体与pSi scrambled,经由脂质体法分别转染至肺癌细胞SPCA1和SH77。通过MTT法检测细胞增殖抑制,用流式细胞仪检测细胞凋亡率和细胞周期,利用半定量RT-PCR和Western blot印迹法分别检测细胞survivin mRNA及蛋白表达水平。结果 Survivin siRNA干预后,SPCA1和SH77细胞均出现凋亡,G0/G1期阻滞。实验组Survivin mRNA水平和蛋白表达水平比脂质体对照组明显减少。结论 Survivin基因特异性RNA干扰可以明显抑制肺癌SPCA1和SH77细胞的体外增殖并诱导明显的细胞凋亡。

Background and objective Survivin,a member of the inhibitor of apoptosis（IAP） protein family,has been demonstrated as a potential new target for apoptosis-based therapy in cancer and lymphoma.The aim of this study is to investigate effects and mechanisms of survivin siRNA transfection on lung adenocarcinoma cell lines SPCA1 and SH77.Methods A siRNA plasmid expression vector and pSi scrambled against survivin were constructed and transfected into SPCA1 and SH77 cells with Lipofectamine 2000.The proliferations of lung adenocarcinoma SPCA1 and SH77 cells were detected by MTT.The apoptotic rate and cell cycle were detected by flow cytometer.The activity of survivin mRNA and protein expression were analyzed with RT-PCR and Western blot.Results Survivin siRNA reduced the proliferation of SPCA1 and SH77 cells.Cell cycle was inhibited in G0/G1.Expressions of survivin siRNA mRNA and protein were reduced in transfected cells compared with the control cells.Conclusion siRNA targeted against survivin can effectively suppress SPCA1 and SH77 cells proliferation and significantly induce SPCA1 and SH77 cells apoptosis.