Survivin特异性siRNA诱导胰腺癌细胞凋亡的研究

Study of p-Survivin-siRNA ExpressionVector on Cell Apoptosis in Pancreatic Cancer Cell Line

目的观察survivin特异性siRNA对胰腺癌细胞株Panc-1细胞增殖和凋亡的影响。方法体外构建Survivin特异性siRNA表达载体p-Survivin-siRNA,转染胰腺癌细胞株Panc-1细胞系,RT-PCR和Western印迹法检验其对胰腺癌细胞株Panc-1细胞的RNA干扰(RNAi)效果。采用MTT法分析其对细胞增殖的影响,流式细胞仪检测其对细胞周期的影响。同时,采用Western印迹法检测半胱氨蛋白酶3(caspase3)的表达变化。结果p-survivin-siRNA表达质粒高效而特异地剔降胰腺癌细胞株Panc-1细胞中survivin的表达,抑制肿瘤细胞增殖(P<0.01),阻断胰腺癌细胞株Panc-1细胞在G1期。随着survivin基因被沉默,caspase3表达升高(P<0.01)。结论靶向survivin基因的siRNA表达载体可以显著抑制胰腺癌细胞株Panc-1细胞增殖,促进细胞凋亡。

Objective To study the effect of Survivin siRNA on proliferation and apoptosis of Panc-1 pancreatic cancer cells. Methods The expressing vector p-Survivin-siRNA containing Survivin siRNA was constructed,, and then transferred into Panc-1 cells by lipofection. The expression of Survivin was detected by RT-PCR and western blotting. The effect of Survivin siRNA on proliferation and apoptosis of Panc-1 cells was determined by MTT test and flow cytometric analysis respectively. Meanwhile, the expression level of caspase 3 was detected by western blotting. Results The expressing vector of Survivin siRNA was observed to efficiently and specifically knockdown the expression of Survivin in Panc-1 cells, and inhibit the growth of Panc-1 cells （P 〈0. 05 ）, and arrest Panc-1 cells in G1 phase. While Survivin gene was silenced, the expression of caspase 3 was increased. Conclusion Survivin siRNA constructed into the expressing vector p-Survivin-siRNA was shown to inhibit proliferation and induce apoptosis of Panc-1 cells.