交联抗人DR5单抗诱导的Jurkat细胞凋亡的信号转导

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对Jurkat细胞增殖的影响，阐明抗DR5抗体的抗肿瘤效应机制。方法MTF方法检测抗体对Jurkat细胞存活率，FrlE—AnnexinV／PI标记流式细胞仪检测交联YM366EC对Jurkat细胞凋亡率影响。交联YM366EC作用于Jurkat细胞2、4、6、8、10、12h收集细胞，提取蛋白Westernblot检测Bcl-2、Cyt—C、Caspase-8、Caspase-3、Bax、Caspase-9的变化。结果抗DR5抗体单独应用不能抑制高表达DR5分子的Jurkat细胞增殖，但应用抗小鼠IgG交联DR5抗体后可直接诱导TRAIL敏感的Jurkat细胞凋亡改变。并且Westernblot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-8、Caspase-3和Caspase-9蛋白的表达增加。结论交联抗DR5抗体YM366EC对Jur—kat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．8、Caspase-3、Caspase-9。

Apoptotic signal transduction of Jurkat cells by cross-linking anti-human DR5 monoclonal antibody

Objective To detect the effect of cross-linking anti-DR5 monoclonal antibody YM366EC on the proliferation of Jurkat cells, and to elucidate the mechanism of anti-tumor by anti-DR5. Methods Cell viability of Jurkat was assayed by MTT. Apoptosis of Jurkat cells induced by cross-linking YM366EC was determined with Annexin V/PI staining using flow cytometry. Bcl-2, Cyt-C, Caspase-8, Caspase-3, Bax and Caspase-9 were determined by Western blot analysis. Results Although inhibition of Jurkat cells were not fund by anti-DR5 monoclonal antibody alone, apoptosis of Jurkat cells were detected with cross-linking anti-DR5 antibody. Cross-linking anti-DR5 YM366EC antibody showed a decreased expression of Bcl-2 and an increased activation of Cyt-C, Bax, Caspase-8, Caspase-3, Caspase-9 by Western blot analysis. Conclusion Apoptosis of Jurkat cells can be induced by cross-linking anti-DR5 YM366EC antibody, and the apoptosis signal pathway is relevant to Cyt-C, Bax, Caspase-8, Caspase-3 and Caspase-9.