交联抗人DR5单抗-YM366EC诱导Jurkat细胞凋亡机制研究.pdfVIP

白慧玲等 交联抗人DR5单抗一YM366EC诱导Jurkat细胞凋亡机制研究 第9期 · 肿瘤免疫学 · 交联抗人DR5单抗．YM366EC诱导Jurkat细胞凋亡机制研究① 白慧玲 杜耀武 王雪垠 李淑莲 王 靖 刘广超 马远方 (河南大学免疫学研究所河南大学细胞与分子免疫学重点实验室，开封475004) 中国图书分类号 R392．1 文献标识码 A 文章编号 1000-484X(2007)09-0789-05 [摘 要] 目的：探讨抗肿瘤坏死因子相关凋亡诱导配体(TRAIL)的死亡受体5(DR5)单抗．YM366EC引起Jurkat细胞 凋亡信号传导通路，阐明抗DR5抗体的抗肿瘤效应机制，为相关肿瘤治疗提供依据。方法：光镜下观察交联366EC作用下 Jurkat细胞形态变化，并利用MqT法检测Jurkat细胞的增殖，利用FITC-Annexin V及PI标记流式细胞仪检测交联YM366EC 对Jurkat细胞凋亡率影响。进一步用Western blot法检测交联YM366EC作用于Jurkat细胞2、4、6、8、10、12小时时Bcl-2、Cyt- C、Bax、caspase3、caspase9的表达变化。结果：DR5抗体YM366EC单独应用无凋亡作用，但应用抗小鼠IgG交联366EC后可致 Jurkat细胞染色质边集、断裂，细胞出芽，凋亡小体形成，并可直接诱导TRAIL敏感的Jurkat细胞凋亡改变。Western blot检测 到随着抗体诱导时间的延长Jurkat细胞Bcl·2蛋白表达减少；Cyt-C、Bax、有活性的caspase3和caspase9蛋白的表达增加。结 论：交联抗DR5抗体YM366EC对Jurkat细胞增殖有明显的抑制作用，诱导的Jurkat细胞凋亡的分子机制涉及到Cyt·C、Bax、 caspase3、caspase9。 [关键词] 交联；Jurkat细胞；抗DR5抗体；凋亡；流式细胞术；免疫印迹 The study on the apoptosis mechanism of the Jurkat cell induced by cross linking anti Dl antibody．YM366EC BAIHui—Ling，DU Yao—Wu，WANG Xue·Yin，LI Shu—Lian，WANG Jing LlU Guang—Chao，MA Yuan—Fang．Research Institute ofImmunology Henan Unwemity，Laboratory of Cell and Molecule Immunology，Henan University，Kalfeng 475004，China [Abstract] Objective：To detect the effect of CIOSS·linking anti·DR5 YM366EC antibody OH the Jurkat cells，and to elucidate the anti tumour mechanism of anti-DR5 antibody．Methods：Morphologic changes of Jurkat cells were observed by a microscope；cytotox· ic and apoptotic effects of cross·linking anti·DR5 to Jurkat cells were detected by MTr and FCM assays．Bcl-2，Cyt-C，caspase3，Bax and caspase9 were detected with Western blot．Results：Chromatin aggregation，budding and apoptotic bodies were seen in Jurkat cells treated by cross-linking anti·DR5 antibody；viability of Jurkat cells was decreased and apoptosis increased obviously．Expression of Bcl- 2，Cyt-C，caspase3，Bax and caspase9 were detected，and they were changed with time of cross·linking anti-DR5 antibody treated．Con· clusion：A