顺铂通过活性氧协同抗hDR5抗体诱导食管癌细胞凋亡  

Cisplatin augments apoptosis of esophageal carcinoma cells induced by agonistic anti-hDR5 antibody by DR5 and mitochondria death pathway

作　　者：刘广超[1,2] 都景芳[1] 刘世贵[2] 龙章富[2] 陶科[2] 高荣[2] 马远方[1] 

机构地区：[1]河南大学细胞与分子免疫学重点实验室,河南开封475000 [2]四川大学生命科学学院生物资源与生态环境教育部重点实验室,四川成都610064

出　　处：《第四军医大学学报》2009年第23期2768-2772,共5页Journal of the Fourth Military Medical University

基　　金：河南省杰出人才创新基金(0321001800)

摘　　要：目的:探讨顺铂与抗死亡受体5(DR5)激动性抗体mDRA-6联合应用对食管癌细胞系EC9706细胞凋亡的影响及作用机制.方法:顺铂、抗体mDRA-6单独或联合作用于食管癌细胞,用流式细胞术检测药物的细胞毒作用、食管癌细胞表面DR5表达、细胞凋亡、细胞内活性氧水平、以及线粒体膜电位的变化;在荧光显微镜下观察细胞凋亡的形态学变化及DR5表达.结果:顺铂和抗体mDRA-6对食管癌细胞均具有细胞毒作用,并具有剂量依赖性,而且顺铂明显提高食管癌细胞对抗体mDRA-6细胞毒的敏感性.顺铂、抗体mDRA-6单独或联合应用均导致食管癌细胞呈现典型细胞凋亡特征.流式细胞术分析结果显示,mDRA-6和顺铂均诱导食管癌细胞凋亡,而且顺铂明显提高细胞对mDRA-6诱导细胞凋亡的敏感性;顺铂增强食管癌细胞表面及胞内的DR5表达,提高细胞内活性氧水平,降低线粒体膜电位.结论:顺铂主要通过活性氧激活细胞内线粒体细胞凋亡信号传导途径以及增强DR5表达,与抗体mDRA-6联合协同诱导食管癌细胞凋亡.研究结果对进一步探讨抗DR5激动性抗体及TRAIL的抗肿瘤临床应用有一定的指导意义.AIM：To probe into the effect of cisplatin in combination with agonistic anti-hDR5 antibody mDRA-6 on apoptosis of human esophageal carcinoma cell line EC9706 and its mechanisms.METHODS：The cytotoxic action of mAb mDRA-6 and cisplatin on EC9706 cells,DR5 expression on EC9706 cell surfaces,apoptosis of EC9706 cells.Mitochondrial membrane potential and reactive oxygen species（ROS） were assessed by flow cytometry.The morphological changes and DR5 expression of EC9706 cells were observed by fluorencence microscope.RESULTS：mDRA-6 or cicplatin exerted cytotoxicity on EC9706 cells in dose-dependent manner,and cisplatin markedly augmented the apoptosis of EC9706 cells induced by mDRA-6.EC9706 cells treated with mDRA-6 or cisplatin exhibited typic apoptostic features in morphology.Apoptosis analysis showed that cisplatin and mDRA-6 could alone induce apoptosis of EC9706 cells and cisplatin sensitized EC9706 cells to mDRA-6 mediated apoptosis.Cytometric analysis revealed that cisplatin could up-regulate DR5 expression and ROS level of EC9706 cells,but decreased mitochondrial membrane potential of EC9706 cells.CONCLUSION：The results present that cisplatin increases DR5 expression on the surfaces of EC109 cells and activates mitochondrial apoptosis pathway by up-regulation of ROS so that cisplatin acts synergistically to induce apoptosis of EC9706 cells in combination with agonistic anti-hDR5 antibody.The results maybe provide potential instruction significance for clinical application of agonistic anti-hDR5 antibody and rTRAIL.

关 键 词：DR5 激动性抗体 TRAIL 细胞凋亡 顺铂 

分 类 号：R73-36[医药卫生—肿瘤]