交联抗DR5单抗YM366EC诱导的Jurkat细胞凋亡机制探讨  

Investigation of the mechanism of apoptosis of the TNF-related apoptosis inducing ligand-susceptible Jurkat cells induced by cross-linking antideath receptor-5 YM366EC

作　　者：白慧玲[1] 杜耀武[1] 王雪垠[1] 李淑莲[1] 王靖[1] 刘广超[1] 马远方[1] 

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

出　　处：《现代免疫学》2007年第4期269-273,共5页Current Immunology

基　　金：河南省杰出人才创新基金(0321001800);河南省医学科技创新人才基金(2002-119)

摘　　要：为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细胞凋亡信号转导通路,阐明抗DR5抗体的抗肿瘤效应机制。观察了交联366EC对Jurkat细胞的形态变化,细胞增殖和细胞凋亡率影响。进一步用Western blot法检测YM366EC作用于Jurkat细胞不同时间Bcl-2、Cyt-C、Bax、Caspase 3、Caspase 9的表达变化。结果发现DR5抗体YM366EC单独应用无凋亡作用,但抗小鼠IgG交联366EC后可致Jurkat细胞染色质边集、断裂,细胞出芽,形成凋亡小体,并可直接诱导TRAIL敏感的Jurkat细胞凋亡。Western blot检测到随着抗体诱导时间的延长Jurkat细胞Bel-2蛋白表达减少;Cyt-C、Bax、有活性的Caspase 3和Caspase 9蛋白的表达增加。结果表明交联抗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、Caspase 3、Caspase 9。To observe the effect of the cross-linking anti-death receptor-5（DRS）YM366EC on the TNF-related apoptosis inducing ligand（TRAIL） and to elucidate the anti-tumor mechanism of their interaction, the morphological changes induced by the cross-linking anti-YM366EC monoclonal antibody were observed microscopically, and its cytotoxic and apoptotic effects were detected with MTT assay and flow cytometry（FCM）. Meanwhile, the changes in the expressions of Bcl-2, Cyt-C, Bax, Caspase-3, Caspase-5 and Caspase-9 after different period of time were further determined by Western blotting. The result showed that after treatment with DR5 antibody YM366EC alone, no apoptotic change could be found, but when Jurkat ceils were treating with the mouse IgG cross-linking with 366EC, chromatin margination, budding and appearance of the apoptotic bodies could be demonstrated in these cells, and it also induced the apoptosis of the TRAIL-susceptible Jurkat cells. As demonstrated by Western blotting analysis, along with the prolongation of the induction time with antibody, the expression of Bcl- 2 protein in Jurkat cells decreased, and those of Cyt-C, Bax, active Caspase-3 and Caspase-9 proteins increased. It is evident that definite inhibitory effect can be demonstrated on Jurkat cells after treatment with cross-linking anti-DR5 antibody YM366EC, and the molecular mechanism of apoptosis induced by this antibody may involve with Cyt-C, Bax, Caspase-3 and Caspase-9.

关 键 词：交联 JURKAT细胞 YM366EC 凋亡 流式细胞术 免疫印迹 

分 类 号：R392.1[医药卫生—免疫学]