siRNA靶向抑制PKR基因及其对宫颈癌Hela细胞凋亡的影响

目的研究siRNA（small interfering RNA,siRNA）对宫颈癌Hela细胞中双链RNA（dsRNA）激活的蛋白激酶（protein kinase regulated by dsRNA,PKR）基因的抑制作用,观察其对dsRNA诱导的细胞凋亡的影响。方法采用RNAi技术,对宫颈癌Hela细胞PKR基因进行特异性抑制;用阳离子脂质体与化学合成的Pre-designed anti-PKR siRNA构建转染复合体,反转染Hela细胞72h后提取总蛋白,用Western blot检测PKR蛋白表达水平的变化,用流式细胞仪检测细胞的凋亡率,激酶活性检测法测定半胱氨蛋白水解酶-3（caspase-3）活性变化。结果 siRNA能降低PKR基因的蛋白表达;未转染组的Hela细胞在加入dsRNA后,细胞凋亡明显增加,caspase-3激酶活性增强（P〈0.05）。与对照组相比,转染组的Hela细胞在加入dsRNA后,细胞凋亡显著减少,caspase-3激酶活性降低（P〈0.05）。结论 siRNA可特异有效地干扰宫颈癌Hela细胞内PKR基因的表达,从而影响肿瘤细胞凋亡。dsRNA可通过激活PKR,增强caspase-3激酶活性,而促进Hela细胞的凋亡。

siRNA can inhibit PKR gene and facilitate apoptosis of Hela cells of uterine cervix cancer:a randomized and controlled trial

Objective To study the inhibitory effects of small interfering RNA(siRNA)of protein kinase regulated by dsRNA(PKR)gene on apoptosis of PKR-related Hela cell line of uterine cervix cancer,and to observe its effect on dsRNA-induced apoptosis.Methods RNA interference(RNAi)was employed to inhibit PKR gene specifically.The transfection compound of cationic lipsome and chemically synthesized Pre-designed anti-PKR siRNA were constructed,and then the total protein was extracted at 72 h after Hela cells were reverse...