RNA干扰双位点抑制Survivin基因诱导雄激素非依赖性前列腺癌PC-3细胞凋亡

目的研究采用RNA干扰技术抑制雄激素非依赖性前列腺癌PC-3细胞中Survivin 基因的表达对PC-3细胞增殖及凋亡的影响。方法设计2条靶向Survivin mRNA的Survivin shRNA,克隆至同一质粒载体,并鉴定分析;质粒转染PC-3细胞,绘制细胞生长曲线;并于转染48 h 后进行Survivin基因蛋白表达及细胞凋亡检测。结果酶切鉴定、测序分析证实负载双片段Sur- vivin shRNA的质粒载体构建成功。转染PC-3细胞后,细胞生长曲线示实验组PC-3细胞增殖明显受抑制;转染后48 h．实验组、阴性对照组、空白对照组Survivin蛋白表达强度分别为(4．62± O．84)％、(38．83±7．96)％和(40．98±3．83)％,实验组与两对照组相比差异均有显著性意义(P< 0．05);转染48 h后PC-3细胞凋亡率开始增加。结论负载双片段Survivin shRNA的质粒载体转染PC-3细胞后能明显抑制PC-3细胞增殖,抑制PC-3细胞Survivin蛋白表达,并能诱导PC-3细胞凋亡。但诱导PC-3细胞凋亡最明显的时机需要进一步观察。

Inhibition of the Survivin gene expression by RNA interference inducing apoptosis of hormone-refractory prostate cancer PC-3 cell line

To investigate the influence of Survivin gene expression inhibited by RNA interference on cell proliferation and apoptosis of hormone-refractory prostate cancer PC-3 cell line. Methods According to the reference, two small hairpair RNA targeting to Survivin mRNA were designed and synthesized, and then inserted into the same plasmid. The sequence of the two small hairpair RNA were testified. After the plasmeds were transfected into PC-3 cell line, cell growth was studied. The survivin protein expression and apoptosis in PC-3 cells was analyzed after transfection for 48 h. Results Enzyme digestion and squencing analyses revealed that the plasmid vectors bearing two small hairpair RNA were constructed successfully. After transfection, PC-3 cell proliferation was inhibited apparently. The expression of Survivin protein was decreased markedly after transfection of the recombinant plamid for 48 h, and the apoptosis rate increased. Conclusion After transfection of the recombinant plasmid loading two shRNA targeting to Survivin mRNA, PC-3 cell proliferation was inhibited, and Survivin protein expression decreased and apoptosis induced.