摘要
目的:构建靶向存活素(survivin)的干扰RNA(siRNA)质粒,观察其对A549细胞增殖、凋亡以及顺铂敏感性的影响。方法:应用pSilencer-U6质粒构建survivin-siRNA干扰质粒,RT-PCR和Western blotting检测survivin mRNA和蛋白的表达,DAPI染色法检测细胞的凋亡,MTT法检测细胞的增殖。结果:成功构建了survivin-siRNA干扰质粒。Survivin-siRNA质粒转染可明显下调A549细胞中survivin mRNA和蛋白的表达、抑制A549细胞的增殖、促进细胞的凋亡、增强A549细胞对顺铂的敏感性。结论:Survivin-siRNA沉默survivin在肺癌细胞中的表达能够抑制肿瘤细胞的增殖、促进细胞凋亡,并能增强肿瘤细胞对化疗药物顺铂的敏感性,survivin可作为肺癌治疗的潜在靶点。
Objective:To investigate the effect of survivin-siRNA plasmid on survivin expression in human lung cancer cell line A549, and to observe its effect on the apoptosis, proliferation, and chemosensitivity of A549 cells. Methods: pSilencer-survivin-siRNA (survivin-siRNA) plasmid was constructed using pSilencer-U6 plasmid and was transfected into A549 cells. Expression of survivin mRNA and protein was examined by RT-PCR and Western blotting analysis, respectively. Apoptosis and proliferation of A549 cells were examined by DAPI staining and MTT, respectively. Results: Survivin-siRNA plasmid was successfully constructed, and it significantly inhibited survivin mRNA and protein expression in A549 cells. Survivin-siRNA transfection induced apoptosis, inhibited proliferation and increased chemosensitivity of A549 cells to cisplatin. Conclusion: pSilencer-survivin-siRNA can silence survivin expression in A549 cells and subsequently inhibit proliferation, promote apoptosis, and enhance chemosensitivity of A549 cells to cisplatin. Survivin may serve as a potential target for gene therapy of lung cancer.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2009年第6期583-587,共5页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.30371624)~~