摘要
目的 为研究脑胶质瘤的耐药、机理及逆转 ,建立了C6耐阿霉素细胞株 (C6/adr)MDR细胞系。方法 采用逐渐增加药物浓度联合反复短期暴露法建立了C6/adr细胞系 ,并对其形态学、超微结构及生长特点进行了研究 ,流式细胞术对细胞周期分布及DNA与RNA含量的变化进行了分析 ,观察了其克隆形成率 ,并对染色体进行了分析 ,用RT PCR法、免疫组织化学及流式细胞免疫学方法研究了多药耐药基因mdr 1及其表达的P gp定性、定量研究 ,用RT PCR法对端粒酶的活性进行了研究 ,建立了该耐药株的动物模型 ,并对其耐药性进行了分析 ,采用MTT法分析了耐药谱。结果 HE染色C6/adr与C6相似 ,透射电镜畸形核多见 ,胞浆内有较多的线粒体、核糖体及滑面内质网 ;群体倍增时间 5 5 2h ;传代 2、 3天 ,C6/adrDNA指数及RNA指数无变化 ;细胞周期分布以G0 +G1期为主 ,克隆形成率可达 5 5 % ;染色体众数多集中于 40～ 42条 ,以亚二倍体居多 ;mdr 1基因表达阳性 ,P gp含量为42 5 1%± 7 82 % ,P gp强阳性 ,主要位于细胞膜上 ,端粒酶呈强阳性 ;动物模型其形态学及P gp含量均无变化 ;该细胞株对ADM ,VM 2 6 ,VCR ,5 Fu ,VP 16 ,CDDP耐药 ,对MTX ,CCNU无耐药性。结论 该耐药细胞系C6/adr耐药表型主要为MDR 1,为研究脑胶质瘤的耐药。
Objective To study drug resistant mechanism and reversal of glioma we established C 6 MDR drug resistant cell line(C 6/adr). Methods This cell line was obtained by sucessive exposure to increasing amount of adriamycin and associated with high dose short explosure of ADM.A series of biological characteristics of C 6 adr were examined, The morphology and microstructure were studied with HE and electron microscope, cell line growth characteristics were studied by MTT. The distribution of cell cycle, the content of DNA and RNA were analysed by FCM. Clone forming ratio and chromosome were observed. Multidrug resistance gene mdr 1 and its encoded P gp were tested quantitatively and qualitatively though RT PCR, immunocytochemical technique and FCM. The telomerase was also examined by RT PCR. C 6/adr SD models were established successfully and their drug resistance was analysed. Meanwhile the sensitivity test of C 6/adr to anticancer drugs were performed with MTT method. Results C 6/adr was the same as C 6 on the morphology(HE), but by transmission electron microscope, there were lots of malformed nuclei, abundant mitochondria and ribosomes, endoplasmic reticular. Populaion doubling time was 55 2 hrs, After C 6/adr generating 2,3 days, DNA and RNA index had no changes; Cell cycle distribution was major in G 0+G 1,clone forming rate was 55%; Chromosomes were on No 40 to 42, most in subdiploid, P gp(42 5±7 82%) and telomerase expressed positive strongly. No changes in morphology (HE) and content of P gp were observed in SD rat model.Drug resistance was to ADM,VM 26,VCR,5F r,VP 16,CDDP,no drug resistance to MTX,CCNU.Conclusions C 6/adr cell line encoded drug resistance gene is MDR 1. This results show that this cell line may be useful in further studying on drug resistance and reversan of glioma.
出处
《中华神经外科杂志》
CSCD
北大核心
2001年第2期109-112,共4页
Chinese Journal of Neurosurgery
基金
国家自然科学基金资助项目!(30 0 70 2 71)
鲁卫攻关课题资助项目!(0 8)
