抗人DR5单克隆抗体对白血病U937细胞凋亡的作用

U937 cell apoptosis induced by anti-human DR5 monoclonal antibody(mDRA-6)

目的:研究鼠抗人DR5单克隆抗体(mDRA-6)对白血病细胞U937的凋亡诱导作用。方法:光镜下观察mDRA-6作用后U937细胞的形态变化;流式细胞术检测U937细胞表面DR5表达率;MTT法检测mDRA-6对U937细胞生长的影响;An-nexinV-FITC/PI双染法流式细胞仪检测细胞凋亡率;琼脂糖凝胶电泳检测U937细胞DNA的片段化降解;JC-1单染流式细胞仪检测细胞线粒体膜电位改变。结果:mDRA-6作用U937细胞后呈现典型的细胞凋亡特征;MTT法检测显示mDRA-6作用U937细胞24小时死亡率为61·09%,并呈时间、浓度依赖性;流式细胞仪检测显示10mg/L的mDRA-6作用4小时,U937细胞凋亡率为79·12%;琼脂糖凝胶电泳显示DNA呈现明显梯状带型;mDRA-6作用U937细胞后线粒体膜电位明显下降。结论:mDRA-6能够诱导白血病U937细胞凋亡,是具有诱导细胞凋亡活性的功能性抗体。

Objective： To evaluate the effects of mouse anti-human DR5 monoclonal antibody （mDRA-6） on the apoptosis of leukemic cell U937.Methods： DR5 expression on leukemic cell surface was detected by flow cytometry with 366EC.The leukemic cell image following mDRA-6 （10 mg/L） treatment for 4 h was evaluated under microscope.The cytoxicity （FCM） induced by mDRA-6 in U937 cells was assayed with MTr method.The apoptosis of U937 after insulting by mDRA-6 （10 mg/L） for4 h or6 h was analyzed by FCM with AnnexinV-F ITC/PI staining and DNA ladder, respectively. Change of mitochondrial membrane potential was detected by FCM with JC-1 staining.Results： U937 cells treated with mDRA-6 （10 mg/L） exhibited typical apoptotic morphology.MTr analysis indicated that the death rate of U937 cells was 61.09% in the presence of 10 mg/L mDRA26 for 24 h.FCM detection indicated that the apoptosis rate of the cells was 79.12% in thre presence of 10 mg/L mDRA-6 for 4 h. Mitochondrial membrane potential declined in the presence of mDRA-6 for 4 h. Conclusion： mDRA-6 is a useful functional antibody with activity of inducing apoptosis of leukemic cell U937.