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目的研究小干扰RNA(small interferingRNA,siRNA)抑制survivin基因的表达对人宫颈癌Hela细胞凋亡的影响。方法根据survivin基因设计序列特异性的siRNA(survivin-siRNA),在脂质体介导下转染Hela细胞,Westernblot方法检测survivin蛋白表达情况,利用TUNEL实验、Annexin-V实验和流式细胞仪方法检测细胞转染前后凋亡的形态学的变化、凋亡率及凋亡周期的变化,Western blot检测caspase-3、caspase-7在转染前后细胞中表达的变化。结果转染survivin-siRNA后,Hela细胞survivin蛋白表达明显下降,与空白对照组及阴性对照组相比,能明显诱导细胞凋亡,细胞被阻滞在G/M期(p<0.05)。转染后Hela细胞中caspase-3、caspase-7蛋2白表达比空白对照组及阴性对照组明显增强(p<0.05)。结论 Survivin-siRNA转染人宫颈癌Hela细胞可上调细胞中survivin蛋白的表达水平,诱导细胞凋亡,与caspase-3、caspase-7的上调有关。
Objective To study the effect of siRNA on the apoptosis of human cervical carcinoma Hela cells. Methods According to survivin gene sequence-specific siRNA (siRNA), Hela cells were transfected by liposome. Western blotting was used to detect the expression of survivin protein. TUNEL assay, Annexin-V assay and flow cytometry The morphological changes of apoptosis, apoptosis rate and apoptosis cycle were detected before and after transfection. The changes of the expression of caspase-3 and caspase-7 in the cells before and after transfection were detected by Western blot. Results The survivin protein expression was significantly decreased in Hela cells transfected with survivin-siRNA. Compared with the blank control group and the negative control group, the apoptosis of Hela cells was significantly induced and the cells were arrested at G / M phase (p <0.05). Transfection of Hela cells caspase-3, caspase-7 protein 2 white expression than the blank control group and negative control group was significantly enhanced (p <0.05). Conclusion Survivin-siRNA transfected human cervical cancer Hela cells can up-regulate the expression of survivin protein and induce cell apoptosis, which is related to the up-regulation of caspase-3 and caspase-7.