阿霉素增强抗人DR5单克隆抗体mDRA-6诱导HL-60细胞凋亡的作用

目的观察抗死亡受体5（DR5）单克隆抗体（单抗）mDRA-6与阿霉素（Adr）对HL-60细胞的协同杀伤作用。方法用DR5蛋白免疫BALB／c小鼠，融合筛选抗DR5杂交瘤细胞，制备抗人DR5单抗mDRA-6；流式细胞术测定Adr对HL-60细胞表面DR5表达的影响；荧光显微镜下观察mDRA-6与Adr协同作用下HL-60细胞的形态变化；MTT法测定1μg／ml Adr与不同浓度的mDRA-6对HL-60细胞存活的影响；琼脂糖凝胶电泳检测mDRA-6与Adr联合对HL-60细胞DNA片断化的影响。结果Adr诱导HL-60细胞表面DR5表达，mDRA-6作用后HL-60细胞出现染色质浓缩、断裂，细胞出芽，凋亡小体形成，mDRA-6与Adr联用细胞形态变化更明显；mDRA-6与Adr对HL-60细胞具有明显的协同杀伤作用，20ng／ml的mDRA-6作用HL-60细胞10h，细胞死亡率为9．32％，1μg／mlAdr作用HL-60细胞10h，细胞死亡率为17.47％，20ng／ml mDRA-6联合1μg／mlAdr，可使HL-60细胞死亡率增至65．06％；20μg／ml mDRA-6与1μg／mlAdr联合作用于HL-60细胞3h，DNA琼脂糖凝胶电泳显示明显“梯形”条带。结论抗DR5单抗mDRA-6与Adr对HL-60细胞具有强大的协同杀伤作用。

Adriamycin enhances anti-human DR5 monoclonal antibody (mDRA-6) induced HL-60 cells apoptosis

Objective To investigate synergistic killing effect of anti-human DR5(death receptor 5 of TRAIL)monoclonal antibody(mDRA-6)and adriamycin(Adr)on HL-60 cells.Methods mDRA-6 was prepared by immunizing BALB/c mice with DR5 protein.DR5 expression on Adr-treated HL-60 cells was de- tected by flow cytometry.Morphologic changes of HL-60 cells were observed under fluorencence microscope. Cytotoxic and apoptotic effects of mDRA-6 and Adr on HL-60 cells were measured by MTT analysis.DNA fragmentation was detected by agarose gel electrophoresis.Results Adr induce DR5 expression on HL-60 cells.Cell budding,chromatin condensation and apoptotic body formation were observed in HL-60 cells trea- led by mDRA-6 and Adr,Death and apoptosis of these cells and DNA ladder were exhibited on agarose gel e- lectrophoresis.Conclusion mDRA-6 and Adr have synergistic killing effect on HL-60 cells.