摘要
目的观察抗死亡受体5(DR5)单克隆抗体mDRA-6对HL-60细胞的凋亡作用。方法制备抗人DR5单克隆抗体mDRA-6;荧光显微镜下观察mDRA-6作用后HL-60细胞的形态变化;MTT法测定不同浓度mDRA-6在不同作用时间对HL-60细胞存活的影响;通过FITC-Annexin V及PI标记细胞,以流式细胞仪检测mDRA-6对HL-60细胞凋亡率的影响;琼脂糖凝胶电泳检测mDRA-6对HL-60细胞DNA片段化的影响。结果mDRA-6导致HL-60细胞染色质浓缩、断裂,细胞出芽,凋亡小体形成;mDRA-6对HL-60细胞具有明显的杀伤作用,24 ng/mL mDRA-6作用HL-60细胞10 h,细胞死亡率达21.2%;1μg/mL的mDRA-6作用8 h,可使HL-60细胞死亡44.1%;Annexin V及PI双染显示,10μg/mL mDRA-6作用2 h,HL-60细胞的凋亡率达48.1%;20μg/mL mDRA-6作用HL-60细胞3 h,DNA琼脂糖凝胶电泳显示明显的“梯形”条带。结论抗DR5单克隆抗体mDRA-6具有诱导HL-60细胞凋亡作用。
Objective To observe apoptotic effect of anti-human DR5 （death receptor5 of TRAIL） monoclonal antibody （mDRA-6） on HL-60 cell. Methods mDRA-6 with different concentrations was used to treat HL-60 cells for differerd times. Morphologic changes of HL- 60 cells treated by mDRA-6 were observed under fluorencence microscope. Cytotoxic and apoptodc effects of mDRA-6 on HL-60 cells were measured by MTT analysis, flow cytometry, and Annexin V-FITC/PI staining. DNA fragmentation was detected by agarose gel electrophoresis. Results Chromatin condensation, budding, and apoptotic bodies were observed in HL-60 cells treated by mDRA-6; Death and apoptosis of HL-60 cells treated by mDRA-6 were increased obviously. ‘DNA ladder＇ in HL-60 cells treated with 20 μg/mL mDRA-6 for 3 h was exhibited on agarose gel electrophoresis. Conclusion Apoptosis of HL-60 cells can be induced by mDRA-6.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2007年第3期295-297,共3页
Immunological Journal
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)资助
