论文部分内容阅读
目的:探讨鼠抗人DR5单克隆抗体(mAb)mDRA-6对Jurkat细胞的细胞毒作用及其机制。方法:以流式细胞术测定mAbmDRA-6对Jurkat细胞的细胞毒作用和细胞凋亡作用,以及caspase8、9的抑制剂对mAbmDRA-6诱导的Jurkat细胞凋亡的影响。在荧光显微镜下,观察mAbmDRA-6对Jurkat细胞形态的影响。以琼脂糖凝胶电泳检测Jurkat细胞中的DNA片段化。结果:mAbmDRA-6对Jurkat细胞具有显著的细胞毒作用,并呈剂量和时间依赖性。经mAbmDRA-6处理后,Jurkat细胞可出现典型的细胞凋亡的形态特征:细胞膜皱缩,出泡,染色质浓缩,形成凋亡小体等。经mAbmDRA-6处理后,Jurkat细胞膜表面高表达丝氨酸磷脂,并可导致Jurkat细胞中的DNA片段化。caspase8的抑制剂可明显抑制mAbmDRA-6诱导的Jurkat细胞凋亡,caspase9的抑制剂的影响很小。结论:mAbmDRA-6可通过死亡受体信号传导途径诱导Jurakt细胞凋亡,对Jurkat细胞产生细胞毒作用,其在以TRAIL/DR5系统进行的肿瘤治疗和探讨DR5功能结构域方面具有广阔的应用前景。
Objective: To investigate the cytotoxic effect of mDRA-6, a murine anti-human DR5 monoclonal antibody (mAb) on Jurkat cells and its mechanism. Methods: The cytotoxicity and apoptosis of mAb mDRA-6 on Jurkat cells were determined by flow cytometry and the effect of inhibitors of caspase 8 and 9 on the apoptosis of Jurkat cells induced by mAbmDRA-6. The effect of mAbmDRA-6 on the morphology of Jurkat cells was observed under a fluorescence microscope. DNA fragmentation in Jurkat cells was detected by agarose gel electrophoresis. Results: mAbmDRA-6 has significant cytotoxicity on Jurkat cells in a dose and time-dependent manner. After treatment with mAbmDRA-6, typical apoptotic morphological features of Jurkat cells are observed: cell membrane shrinkage, effervescence, chromatin condensation, formation of apoptotic bodies and the like. After treatment with mAbmDRA-6, serine phospholipids were highly expressed on the surface of Jurkat cell membranes and could lead to DNA fragmentation in Jurkat cells. Inhibitors of caspase8 significantly inhibited the apoptosis of Jurkat cells induced by mAbmDRA-6, and the effect of caspase9 inhibitors was small. CONCLUSION: mAbmDRA-6 can induce Jurakt cell apoptosis via death receptor signaling pathway and induce cytotoxic effect on Jurkat cells. It has broad application prospect in the treatment of tumors with TRAIL / DR5 system and exploration of DR5 functional domains .