JNK信号传导通路在抗人DR5单克隆抗体mDRA-6诱导白血病细胞凋亡中的作用  

The role of c-jun N-terminal Kinase (JNK) signaling pathway in the apoptosis of leukemic cells induced by anti-human DR5 monoclonal antibody mDRA-6

作　　者：蔡静[1] 李淑莲[1] 王靖[1] 王赟[1] 袁艳军[1] 蒋祺[1] 秦方园[1] 马远方[1] 

机构地区：[1]河南省天然药物与免疫工程重点实验室河南大学免疫学研究所,开封475004

出　　处：《中国免疫学杂志》2009年第10期916-919,共4页Chinese Journal of Immunology

基　　金：国家863计划(2006AA02A254);河南省杰出人才创新基金(074200510014)资助

摘　　要：目的:探讨JNK信号传导通路在抗人DR5单克隆抗体mDRA-6诱导白血病细胞Jurkat和U937凋亡过程中的作用。方法:MTT法检测mDRA-6对Jurkat和U937细胞的生长抑制作用;以Caspase抑制剂Z-VAD-FMK和JNK抑制剂SP600125进行干预,采用Annexin V-FITC/PI双染流式细胞术定量分析Jurkat和U937细胞凋亡;利用免疫印迹技术检测mDRA-6作用下凋亡细胞的JNK蛋白的表达情况。结果:mDRA-6对Jurkat和U937细胞具有明显的生长抑制作用;Annexin V-FITC/PI双染流式细胞术检测显示,10μg/ml的mDRA-6作用Jurkat和U937细胞3小时,其凋亡率分别为62.62%和35.65%,JNK抑制剂干预后细胞凋亡明显增加;免疫印迹技术检测显示,mDRA-6作用细胞5分钟后JNK即呈现活性片段表达,并随作用时间的延长其活性增强,而Caspase全抑制剂干预后,5、15分钟JNK磷酸化增强,但随后随时间递减。结论:mDRA-6抑制Jurkat和U937细胞生长并诱导其凋亡,其凋亡过程中有JNK的激活并发挥抗凋亡作用。Objective： To explore of the role of JNK/SAPK signal transduction system in the apoptosis of hurman leukemic cell lines Jurkat and U937 induced by anti-human DR5 monoclonal antibody mDRA-6. Methods： Human leukemic cell lines Jurkat and U937 were treated with different concentrations of mDRA-6. MTT assay was used to observe the proliferation of Jurkat and U937cells, and Flow cytomctry was used to examine apoptosis rate before and after JNK signal was blocked by SP600125, a JNK inhibitor. The expression of p-JNK was detected by Western blot. Results： mDRA-6 inhibited the proliferation of Jurkat and U937 cells in a concentration- and time-dependent manner. Flow cytometry showed that mDRA-6 induced the apoptosis of Jorkat and U937 cells, while the apoptosis rate was increased after JNK signal was blocked by SP600125, a JNK inhibitor. Western blot exhibited that mDRA-6 induced the activation of p-JNK. The expression of p-JNK was detected 5 minutes after mDRA-6 treatment, while the activation of p-iNK was gradually suppressed after Caspase inhibitor Z-VAD-FMK treatment. Con- clusion： mDRA-6 can inhibit the proliferation and induce the apoptosis of leukemic cell lines Jurkat and U937 in vitro, mDRA-6-induced apoptosis activated p-JNK,SP600125 inhibits the activation of JNK, and therefore increases the apoptosis rate of Jurkat and U937 cells induced by mDRA-6.

关 键 词：JNK 抗死亡受体5单克隆抗体 白血病细胞 凋亡 

分 类 号：R392.1[医药卫生—免疫学]