siRNA沉默SIRT1基因对宫颈癌细胞HeLa增殖和凋亡的影响

化学合成靶向SIRT1基因的小干扰RNA,脂质体法转染人宫颈癌细胞株HeLa,观察小干扰RNA沉默SIRT1基因对HeLa增殖及细胞凋亡的影响。在优化siRNA SIRT1转染条件的基础上,应用RT-PCR和Western blot分别检测各组SIRT1 mRNA、SIRT1蛋白及凋亡相关蛋白的表达;CCK-8法检测细胞增殖抑制率;Hoechst荧光染色法和流式细胞仪检测细胞凋亡。结果表明,siRNA SIRT1转染细胞组SIRT1 mRNA水平和蛋白表达量明显低于对照组;siRNA SIRT1转染组细胞增殖受抑制,细胞凋亡率明显增加;凋亡相关蛋白P53、P21表达上调,Survivin表达下调。上述结果表明:siRNA SIRT1诱导的HeLa细胞凋亡与P53、P21、Survivin通路关系密切,但siRNA SIRT1诱导HeLa细胞凋亡的详尽机制有待进一步研究。

Effect of Small Interference RNA Silent SIRT 1 Gene on Proliferation & Apoptosis of Cervical Cancer Cell Line HeLa

The effects of mini interference RNA of chemosynthetic target oriented SIRT1 gene on the proliferation and apoptosis of human cervical cancer cell line HeLa were investigated. Based on optimized siRNA SIRT1 transfection condition, the level of SIRT1 mRNA, SIRT1 protein and apoptosis related protein P53, P21, survivin were measured by RT-PCR and Western blot respectively, the cell proliferation inhibition rate was assessed by CCK-8, and the apoptosis was evaluated by Hoechst fluorochrome staining and flow cytometry. The results showed that in the siRNA SIRT1 transfection group the SIRT1 mRNA and protein expression was significantly lower than that in the control groups, the growth of HeLa cells was inhibited and the cell apoptosis rate was increased significantly. The apoptosis related protein P53, P21 increased their expression, and survivin decreased its expression. These results indicated that siRNA SIRT1 induced apoptosis in HeLa cells closely related to P53, P21 and survivin paths. However, further studies remain to investigate the detailed mechanism of siRNA SIRT1 induced apoptosis in HeLa cells.