抗DR5单抗增强顺铂诱导HeLa细胞凋亡作用的研究  

Anti-DR5 monoclonal antibody enhances cisplatin-inducing apoptosis of HeLa cells

作　　者：赵粤萍[1] 贾彩云[1] 马远方[1] 沈倍奋[1] 

机构地区：[1]河南大学免疫学研究所河南大学细胞与分子免疫学实验室,开封475004

出　　处：《现代免疫学》2007年第3期198-201,共4页Current Immunology

基　　金：国家自然科学基金(30571697);河南省杰出人才创新基金(0321001800)

摘　　要：肿瘤坏死因子相关凋亡诱导配体(TNF-related apoptosis-inducing ligand,TRAIL)主要通过死亡受体5(death recep- tor,DR5)诱导肿瘤细胞凋亡。本文旨在探讨顺铂对HeLa细胞表面DR5分子表达影响及抗DR5单抗mDRA-6对顺铂诱导HeLa细胞凋亡增强作用。用间接免疫荧光染色结合流式细胞术分析DR5分子表达;MTT法检测HeLa细胞毒作用;用AnnexinV/PI双染试剂盒检测细胞凋亡率;荧光显微镜观察凋亡细胞的形态学改变。结果:正常HeLa细胞表面DR5表达量为30.01%,顺铂不能上调HeLa细胞表面DR5表达;mDRA-6可以明显提高顺铂对HeLa细胞的细胞毒作用,且存在剂量效应关系。IC_(50)值约为12.5μg/ml。研究结果表明,mDRA-6能够明显增强顺铂对HeLa细胞的细胞毒及细胞凋亡作用。The TNF- related apoptosis-inducing ligand （TRAIL） induces apoptosis of tumor cells mainly via death receptor- 5 （DRS）. The aim of this study is to investigate the influence of cisplatin on the expression of DR5 on the surface of HeLa cells and the enhancing effect of anti-DR5 apoptotic monoclaonal antibody mDRA-6 on the cisplatin-inducing apoptosis of HeLa cells. The expression of DR5 on HeLa cells was detected by indirect immuno-fluorescence assay （IFA） and flow cytometry（FCM）. Meanwhile, the cytotoxicity of HeLa cells was examined by MTT assay, apoptosis was determined by FCM with Annexin V/ PI staining and the morphological chracteritics of the apoptotic cells was observed under digital high-sensitivity fluorescence microscopy. It was found that the expression percentage of DR5 on the surface of normal HeLa cells was 30.01%. Although cisplatin was unable to up-regulate this kind of expression, but the anti-DR5 apoptotic monoclonal antibody mDRA-6 could enhance remarkably the cisplatin-inducing apoptosis of HeLa cells with a concentration-dependent cytotoxicity, in wich the IC50 was proved to be 12.5 μg/ml, It is concluded that the anti-DR6 apoptotic monclonal antibody mDRA-6 can enhance the cisplatin-inducing apoptosis in HeLa cells.

关 键 词：MDRA-6 DR5 顺铂 HeLa细胞 细胞毒 凋亡 

分 类 号：R392.5[医药卫生—免疫学]