TRAIL联合顺铂诱导卵巢癌耐药细胞株COC1/DDP凋亡机制的研究

摘要

Objective:To investigate the effect of TRAIL combined with Cisplatin on the growth of ovarian cancer resistant COC1/DDP cells,and the effect of drug combination on the expression of TRAIL death receptors(DR4,DR5) and the apoptosis related genes Smac and Survivin.Method:The effects of different concentrations of TRAIL protein combined with DDP on the growth inhibition of COC1/DDP cells were detected by the MTT assay.Apoptosis of COC1/DDP cells was detected by V-FITC Annexin method.The expression of TRAIL death receptors(DR4,DR5) and the apoptosis related genes Smac,Survivin after treated with DDP were studied by RT-PCR technique.Result:TRAIL protein inhibited the growth of COC1/DDP cells,and the inhibition rate of DDP and TRAIL protein was significantly increased(P<0.05).The COC1/DDP cell expression level of DR5 significantly enhanced as the control group of 3.45 times(P<0.001).The expression of Smac for the control group of 2.82 times(P<0.001).DR4 level without obvious change.Survivin expression compared with the control group decreased significantly(P<0.001).Conclusion:TRAIL protein inhibited the growth of COC1/DDP cells,and the combination of DDP and TRAIL enhanced the inhibition of COC1/DDP cells.TRAIL reversing the drug resistance of COC1/DDP cells to DDP may cause by the increase of TRAIL receptor DR5 and Smac expression,and promote the apoptosis of tumor cells through the mitochondrial pathway.%目的：研究TRAIL联合顺铂对卵巢癌耐药细胞COC1/DDP生长的影响，以及联合用药对TRAIL死亡受体（DR4、DR5）、凋亡相关基因Smac、Survivin表达的影响，揭示TRAIL联合顺铂可能逆转COC1/DDP细胞耐药性的机制。方法：采用MTT法检测不同浓度TRAIL蛋白与DDP联合用药对COC1/DDP细胞生长的影响，用Annexin V-FITC法检测COC1/DDP细胞凋亡，用RT-PCR方法检测DDP对TRAIL受体（DR4、DR5）、凋亡相关基因Smac、Survivin表达。结果：TRAIL蛋白对COC1/DDP细胞生长有抑制作用，DDP与TRAIL蛋白联合作用后细胞生长抑制率显著升高（P<0.05）。DDP使COC1/DDP细胞的DR5表达水平显著增强，为正常对照组的3.45倍（P<0.001）；Smac表达为对照组的2.82倍（P<0.001）；DR4水平无明显变化；Survivin表达较对照组显著减少（P<0.001）。结论：TRAIL蛋白对COC1/DDP细胞生长有抑制作用，DDP与TRAIL联合增强了对COC1/DDP细胞生长抑制；TRAIL逆转COC1/DDP细胞对DDP的耐药性可能与DDP导致TRAIL受体DR5水平升高、Smac表达增加，通过线粒体途径促进了肿瘤细胞的凋亡有关。