【摘要】 目的:研究p38MAPK在卵巢上皮癌顺铂化疗耐药中的作用,并探讨其作用机制。方法:蛋白质印迹法检测顺铂对卵巢癌中p38MAPK的激活情况;MTT法检测p38MAPK抑制剂SB203580处理后及p38shRNA干扰后细胞顺铂耐药指数的变化;实时定量PCR技术检测SB203580处理后及p38shRNA干扰后,耐药蛋白ERCC1、MDR、LRP、GST-π及凋亡蛋白Caspase-3、Survivin等mRNA的表达变化。结果:在一定时间浓度梯度下,顺铂可诱导卵巢癌顺铂敏感株COC1及耐药株COC1/DDP细胞内p38MAPK的激活,但耐药株的激活程度弱于敏感株;p38MAPK抑制剂SB203580及p38shRNA干扰可增加卵巢癌细胞株的耐药指数(t=4.610,P=0.041;t=11.621,P=0.000);SB203580及p38shRNA干扰后SurvivinmRNA(t=5.152,P=0.007;t=6.008,P=0.004)、ERCC1mRNA(t=13.742,P=0.005;t=11.621,P=0.000)及LRPmRNA(t=8.173,P=0.001;t=16.815,P=0.000)的表达明显上调。结论:p38MAPK受抑制可能导致卵巢上皮性癌顺铂耐药的产生,其机制可能是通过上调Survivin、ERCC1及LRP的mRNA表达来实现。更多还原

【Abstract】 OBJECTIVE:To investigate the role of p38MAPK in cisplatin resistant ovarian cancer and to explore its mechanism.METHODS:Using Western blot to detect the activation of p38MAPK in ovarian cancer treated with cisplatin;Using MTT assay to detect the change of cisplatin resistant index after adding p38MAPK inhibitor SB203580 or disturbed by p38 shRNA;Real-time quantitative PCR was used to detect the change of resistance protein ERCC1,MDR,LRP,GST-π and the apoptotic protein Caspase-3,Survivin mRNA’s after adding p38MAPK inhibitor SB203580 or disturbed by p38 shRNA.RESULTS:At a certain time/concentration gradient,cisplatin could activate p38MAPK both in ovarian cancer cisplatin-sensitive line COC1 and cisplatin-resistant line COC1/DDP cell but the activation level of COC1/DDP cell was weaker;SB203580 and p38 shRNA could increse the RI(Resistant Index) of ovarian cancer cell lines(t=4.610,P=0.041;t=11.621,P=0.000);Survivin mRNA(t=5.152,P=0.007;t=6.008,P=0.004),ERCC1 mRNA(t=13.742,P=0.005;t=11.621,P=0.000)and LRP mRNA(t=8.173,P=0.001;t=16.815,P=0.000) were significantly upregulated after treating with SB203580 or p38 shRNA.CONCLUSION:Inhibition of p38MAPK may result in cisplatin resistance in epithelial ovarian cancer cells and it is probably caused by the up-regulation of the mRNA expression of Survivin,ERCC1 and LRP.更多还原