摘要
应用Lipofectin介导的基因转移技术,将人MDR1-cDNA导入EJ膀胱癌细胞,经60、120和240μg/L秋水仙素的逐级筛选,挑出两个耐药的克隆细胞系,分别培养在240μg/L或480μg/L秋水仙素的培养基中,命名为EJ-R240或EJ-R480。对照组EJ膀胱癌细胞在60μg/L秋水仙素的培养基中不能生存。Southen、RNA斑点杂交分析和免疫组化分析显示,人MDR_1-cDNA已整合在转染细胞的基因组中,并有相应的mRNA和蛋白产物P-糖蛋白的表达。药敏试验结果显示EJ-R240和EJ-R480对秋水仙素的耐受性分别是对照组的73.6和84.2倍。资料表明,MDR_1基因及其产物P-糖蛋白的过度表达可赋予转染细胞的抗药表型。因此,本研究不仅从基因水平上证实MDR_1基因及P-糖蛋白的异常表达可能是膀胱癌耐药性产生的一个重要途径或机制,同时也为研究MDR_1基因介导的多项耐药以及寻找有效的逆转途径提供了一个理想的实验模型。
Human multidrug resistance(MDR1)gene was introduced into EJ bladder carcinoma cells by lipofectin-mediated gene transfer.The transfected cells were selected by stepwise increase of colchicine concentration(60,120 and 240μg/L).Finally,two drug resistant clones,growing stably in complete 1640 medium containing 240μg/L and 480μg/L colchcine,were obtained and were designated as EJ-R240 and EJ-R480.On the other hand,the control EJ cells could not live in the complete 1640 medium containing even 60μg/L colchicine.The southern, RNA dot hybridization and immunohistochemistry assay designated that human MDR1 cDNA was integraded into genome of the transfected cells.The drug sensitivity test indicated that resistance of EJ-R240 and EJ-R480 to colchicine has been 73.6 and 84.2 fold higher than the control parental cells. The result indicated that over expression of MDR1 gene and P -glycoprotein could confer the drug resistance on bladder carcinoma cells.It was claimed that such transfected cells might serve as an ideal model to study MDR1 mediated multidrug resistance and to search the effective means to reverse MDR1 gene mediated multidrug resistance.
出处
《中华泌尿外科杂志》
CAS
CSCD
北大核心
1996年第4期203-206,共4页
Chinese Journal of Urology
关键词
膀胱肿瘤
抗药性
耐药基因
Bladder neoplasms Drug tolerance Genes