亚毒性剂量阿霉素提高抗人DR4、DR5抗体诱导的神经胶质瘤细胞凋亡的研究
Enhancement of glioma cell apoptosis induced by anti-human DR5/DR4 monoclonal antibodies by sub-toxic dose of doxorubicin in human
摘要
目的探讨亚毒性剂量的阿霉素影响抗DR4、DR5单克隆抗体(mAb)FMU1.4和FMU1.5诱导3株神经胶质瘤细胞株U343(TRAIL敏感株)、U138(TRAIL部分敏感株)及U373(TRAIL耐受株)凋亡的作用及可能的机制。方法采用流式细胞术检测DR4、DR5的表达及神经胶质瘤细胞中DNA倍增。用MTT比色法检测mAbFMU1.4和FMU1.5对3株神经胶质瘤细胞增殖的抑制作用。用共聚焦显微镜观察3株细胞中Ca2+的浓度。以Westernblot检测细胞内色素C、FLIP[FLICE-inhibitoryprotein,为一组含有死亡效应结构域(DED)的胞浆蛋白]的表达。结果亚毒性剂量的阿霉素作用后,DR5、DR4在3株细胞中的表达提高;而mAbFMU1.4、FMU1.5诱导U138和U373细胞凋亡的作用增强,细胞内细胞色素C的表达提高,FLIP的表达降低,Ca2+浓度增加。结论亚毒性剂量的阿霉素与抗DR4、DR5mAb联合应用后,可提高mAb诱导靶细胞凋亡的效应,其作用机制与DR4、DR5、细胞色素C、FLIP的表达及Ca2+的含量有关。
【英文摘要】 AIM: To study the cytotoxic effects of doxorubicin on apoptosis in glioma cell lines U343, U138, U373 induced by anti-human DR4/DR5 monoclonal antibodies (FMU1.4/FMU1.5) and the underlying mechanism. METHODS: Expression of DR4/DR5 was quantitated by flow cytometry. Cytotoxicity exerted by FMU1.4/FMU1.5 on three cell lines was measured by MTT colorimetry and the induced apoptosis was determined by agarose gel electrophoresis. The expression of cytochrome C, FLIP and Ca 2+ concentration were also measured. RE...