FLIP蛋白抑制顺铂诱导大鼠C6胶质瘤细胞凋亡  

FLIP inhibits induction of apoptosis by cisplatin in rat C6 glioma cells

作　　者：贺亚龙[1] 贺晓生[1] 章翔[1] 屈朔瑶[2] 王江[1] 

机构地区：[1]第四军医大学西京医院神经外科,陕西西安710032 [2]第四军医大学西京医院呼吸内科,陕西西安710032

出　　处：《中华神经外科疾病研究杂志》2009年第4期316-319,共4页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(30672371);陕西省自然科学基金资助项目(2005C209)

摘　　要：目的探讨自杀相关因子(Fas)相关死亡结构域样白介素1(IL-1)β转化酶抑制蛋白(FLIP)对于顺铂(CDDP)诱导大鼠脑胶质瘤细胞(C6细胞株)凋亡的抑制作用,为进一步研究胶质瘤的耐药性奠定分子生物学基础。方法利用由Ad-Max腺病毒包装系统成功构建的携载大鼠FLIP基因的腺病毒表达载体Ad-FLIP感染大鼠C6胶质瘤细胞,24h后经逆转录酶-多聚酶链反应(RT-PCR)及Western blot检测感染组及对照组细胞中FLIP基因的mRNA及蛋白表达水平;分别给予Ad-FLIP感染组及对照组细胞不同浓度的CDDP(0,1,2,4,8mg/ml),药物处理48h后,经流式细胞仪(FCM)分析细胞凋亡状况;四唑蓝显色法(MTT)测定并比较两组细胞活力。结果Ad-FLIP感染组细胞与对照组细胞相比,FLIP mRNA和蛋白表达水平明显增高;流式细胞仪检测结果显示Ad-FLIP感染组细胞凋亡率明显低于对照组。MTT法结果提示经CDDP处理后,Ad-FLIP感染组与对照组细胞活力均有下降,但FLIP蛋白具有明显的抑制作用。结论FLIP蛋白在大鼠C6胶质瘤细胞中具有抵抗化疗药物的作用。Objective To explore the effect of Fas-associated death domain-like interleukin-1 betaconverting enzyme-inhibitory protein （FLIP） on apoptosis of brain glioma C6 cell lines induced by cisplatin （CDDP）. Methods Recombined adenovirus Ad-FLIP was constructed by Ad-max system. The C6 glioma cells were transfected with Ad-FLIP, The expression of FLIP was detected by the techniques of reverse transcriptasepolymerase chain reaction （RT-PCR） and Western blot. When infected C6 glioma cells and control cells were treated with CDDP at different concentrations （0, 1, 2, 4, 8 mg/ml ） for 48 hours, apoptotic peaks were identified by flow cytometrle study. Meanwhile the cell viability was measured by methyl thiazolyl tetrazolium assay （MTT）. Results After transfected with the adenovirus vector carrying FLIP gene into brain glioma （26 cells, mRNA and protein level of FLIP elevated markedly. FLIP obviously suppressed the apoptosis induced by different eoneentrations of CDDP. The cell viability was elevated by Ad-FLIP. Conclusion FLIP can inhibit the brain glioma C6 cell apoptosis induced by CDDP, so FLIP might be considered as a target gene for treatment of glioma.

关 键 词：FLIP 胶质瘤 凋亡 顺铂 

分 类 号：R739.4[医药卫生—肿瘤]