抗人DR4、DR5单克隆抗体诱导神经胶质瘤细胞凋亡的实验研究

Anti-human DR5/DR4 monoclonal antibody induce glioma cell lines apoptosis

目的:探讨抗死亡受体DR4、DR5单克隆抗体(mAb)FMU1·4、FMU1·5对3株神经胶质瘤细胞株U343(敏感株)、U138(部分敏感株)、U373(耐受株)的杀伤作用及机制。方法:采用流式细胞术、RT-PCR、免疫细胞化学法、MTT比色法、电泳、DNA倍体分析和Westernblot等方法。结果:DR5在U343高表达;DR4在U373低表达,U343对FMU1·5敏感并呈剂量依赖性、对FMU1·4部分敏感;U138对FMU1·5部分敏感,对FMU1·4耐受;U373对两种抗体耐受。结论:FMU1·4、FMU1·5能不同程度地诱导三株细胞凋亡,其机制与DR4、DR5、细胞色素C和FLIP的表达有关。

Objective：To study the cytotoxic effects on three glioma cell lines U343, U138, U373 induced by anti-human DRS/DR4 moncelonal antibodies（FMU1.5/FMU1.4） and the underlying mechanism. Methods：Expression of DR4/DR5 was quantitated by flow cytometry and DIL/4DR5 mRNA detected by RT-PCR. Cytotoxicity exerted by FMU1.4/FMU1.5 on three cell lines was measured by MTr colorimetry and the induced apoptosis was determined by agarose gel electrophoresis, DNA ploldy analysis was studled by flow cytometry. Results：Tbe expression of DR5 on U343 cells was higher and the expression of DR4 on U373 cells was lower. Cell line U343 was sensitive to FMU1.5 and in a dose dependent manner, but it was partially sensitive to FMU1.4; Cell line U138 was partially sensitive to FMU1.5 and resistant to FMU1.4; Cell line U373 was insensitive to two antibodies. Conclusion：Apoptosis induced by moncelonal antibedie FMU1.4/FMU1.5 vary among three cell lines. The underlying mechanism may be relevant to DR4/ DR5 expression, the release of cytcehrome C and FLIP.