抑制survivin基因的表达对骨肉瘤细胞株U-2OS凋亡的影响
| 作者: |
1张文韬,
1段宁,
1程辉光,
1宋涛,
1杨团民,
1同志超
1 西安市红会医院骨科，西安 710054 |
| 通讯: |
张文韬
Email: zhangwentao1975@126.com |
| DOI: | 10.3978/j.issn.2095-6959.2015.05.028 |
| 基金: | 西安交通大学医学院附属红会医院基金, YJ2013012 |
摘要
目的：观察抑制survivin基因表达后对骨肉瘤细胞株U-2OS凋亡的影响。方法：下调骨肉瘤细胞株U-2OS中survivin的表达，并采用免疫印迹法(western blot)和逆转录聚合酶链反应(RT-PCR)检测U-2OS细胞株中survivin的蛋白和mRNA表达；同时采用甲基偶氮哇盐(MTT)法、流式细胞术和western blot检测下调survinin对化疗药物顺铂增加细胞凋亡和减少存活率作用的影响，并探讨其对抗凋亡分子Bcl-2以及促凋亡分子Bax的蛋白表达的作用。结果：survivin-siRNA成功转染U-2OS细胞株后，western blot和RT-PCR检测结果均显示U-2OS细胞株中survivin的蛋白和mRNA表达水平显著低于空白对照组(P<0.05)；MTT比色法结果显示下调survivin可促进化疗药物顺铂对U-2OS细胞株存活率降低及生长抑制率增加的作用；western blotting检测结果表明，与空白对照组相比，Bcl-2的蛋白表达显著减少(P<0.05)，而Bax的蛋白表达显著增加(P<0.05)。结论：下调survivin基因表达可促进骨肉瘤U-2OS细胞株的凋亡，并增加其对化疗药物顺铂的敏感性，为以survivin作为靶基因治疗骨肉瘤提供实验依据。
关键词：
survivin
siRNA
肉骨瘤细胞株U-2OS
化疗敏感性
Blockade of survivin promotes the apoptosis of human osteosarcoma cell line U-2OS
CorrespondingAuthor: ZHANG Wentao Email: zhangwentao1975@126.com
DOI: 10.3978/j.issn.2095-6959.2015.05.028
Abstract
Objective: To investigate the effect of surviving on the apoptosis of human osteosarcoma cell line U-2OS. Methods: After transfection with small inference RNA targeting survivin in U-2OS, survivin protein and mRNA expression were measured with western blot and RT-PCR, respectively. U-2OS cells were knockdown with survivin, and then were incubated with cisplatin for the indicated times, cell survival rate, growth inhibitory rate and apoptosis rate were measured with MTT and flow cytometry, respective; Bcl-2 and Bax protein expressions were measured with western blot. Results: Knockdown of survivin significantly decreased the protein and mRNA expression of survivin (P<0.05). Furthermore, survivin deficiency dramatically augmented the effect of cisplatin on cell survival rate, growth inhibitory rate, apoptosis rate, and the expression of Bcl-2 and Bax (P<0.05). Conclusion: Absence of survivin accelerated the apoptosis of U-2OS, and increased the chemosensitivity of U-2OS towards cisplatin; the present study reveals a novel role of surviving in the treatment of osteosarcoma.