CAJ | 학술논문

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。
목적：관찰억제survivin기인표체후대골육류세포주U-2OS조망적영향。방법：하조골육류세포주U-2OS중survivin적표체，병채용면역인적법(western blot)화역전록취합매련반응(RT-PCR)검측U-2OS세포주중survivin적단백화mRNA표체；동시채용갑기우담왜염(MTT)법、류식세포술화western blot검측하조survinin대화료약물순박증가세포조망화감소존활솔작용적영향，병탐토기대항조망분자Bcl-2이급촉조망분자Bax적단백표체적작용。결과：survivin-siRNA성공전염U-2OS세포주후，western blot화RT-PCR검측결과균현시U-2OS세포주중survivin적단백화mRNA표체수평현저저우공백대조조(P<0.05)；MTT비색법결과현시하조survivin가촉진화료약물순박대U-2OS세포주존활솔강저급생장억제솔증가적작용；western blotting검측결과표명，여공백대조조상비，Bcl-2적단백표체현저감소(P<0.05)，이Bax적단백표체현저증가(P<0.05)。결론：하조survivin기인표체가촉진골육류U-2OS세포주적조망，병증가기대화료약물순박적민감성，위이survivin작위파기인치료골육류제공실험의거。
Objective:To investigate the effect of surviving on the apoptosis of human osteosarcoma cell line U-2OS. Methods:Atfer 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 lfow 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 deifciency 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.