RNA干扰抑制生存素表达并诱导PC-3细胞凋亡

[RNA interference inhibits the expression of survivin and induces the apoptosis of PC-3 cells].

作者信息

Wang Yi-bing, Liu Yi-li, Wang Ping, Fu Xiao-liang, Sun Zhi-xi

机构信息

Department of Urology, the Fourth Hospital Affiliated to China Medical University, Shenyang, Liaoning 110032, China.

出版信息

Zhonghua Nan Ke Xue. 2007 Nov;13(11):983-7.

PMID:18077908

Abstract

OBJECTIVE

To investigate the inhibitory effect of RNA interference (RNAi) on the expression of survivin mRNA and its inducibility of the apoptosis of PC-3 cells.

METHODS

siRNA expression vectors were designed and constructed to be directed at survivin and transfected into PC-3 cells by Lipofectamine in 4 groups: plasmid A, plasmid B, negative sequence and control E. RT-PCR, Western blot and flow cytometry were used to detect the expression of survivin mRNA and the apoptosis of PC-3 cells.

RESULTS

The expression rates of survivin protein were 18.94% +/- 0.63%, 16.35 +/- 0.23%, 46.41% +/- 0.76% and 46.20 +/- 1.47%, those of survivin mRNA were 27.94% +/- 1.43%, 24.51% +/- 1.37%, 49.46% +/- 0.71% and 48.49% +/- 1.32%, and the apoptosis rates of PC-3 cells were 12.80% +/- 1.33%, 16.48% +/- 1.00%, 3.03% +/- 0.62% and 2.96% +/- 0.41% respectively in different groups.

CONCLUSION

RNAi can effectively inhibit the expression of survivin mRNA and induce the apoptosis of PC-3 cells.

摘要

目的

探讨RNA干扰（RNAi）对survivin mRNA表达的抑制作用及其诱导PC-3细胞凋亡的能力。

方法

设计并构建针对survivin的小干扰RNA（siRNA）表达载体，采用脂质体法转染PC-3细胞，分为质粒A组、质粒B组、阴性序列组和对照组E。运用逆转录-聚合酶链反应（RT-PCR）、蛋白质免疫印迹法（Western blot）和流式细胞术检测survivin mRNA的表达及PC-3细胞的凋亡情况。

结果

不同组中survivin蛋白表达率分别为18.94%±0.63%、16.35±0.23%、46.41%±0.76%和46.20±1.47%；survivin mRNA表达率分别为27.94%±1.43%、24.51%±1.37%、49.46%±0.71%和48.49%±1.32%；PC-3细胞凋亡率分别为12.80%±1.33%、16.48%±1.00%、3.03%±0.62%和2.96%±0.41%。