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

作　　者：庄国洪[1] 宋玉国[2] 毕胜利[2] 杜柏榕[3] 朱迅[3] 

机构地区：[1]厦门大学医学院抗癌研究中心,福建厦门361005 [2]北华大学医学院临床免疫研究室,吉林吉林132011 [3]吉林大学基础医学院免疫学教研室,吉林长春130021

出　　处：《细胞与分子免疫学杂志》2007年第2期120-123,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：教育部跨世纪优秀人才培养计划资助项目(2000年)

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。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 （FMUI. 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. RESULTS： Following the treat- ment of doxorubicin DR4 and DR5 were highly expressed on the cell surface; The apoptosis of U138 and U373 induced by FMUI. 4 and FMU1.5 was stronger, expression of cytochrome C and Ca^2＋ concentration were enhanced, whereas the expression of FLIP was downregulated. CONCLUSION： Subtoxic doxorubicin applied with antibodies caused higher cell death rate of glioma cells, which may be relevant to DR4/DRS, the release of cytochrome C and FLIP and Ca^2 ＋ concentration.

关 键 词：TNF相关凋亡诱导配体 死亡受体 凋亡 阿霉素 

分 类 号：R392.12[医药卫生—免疫学]