转基因法建立膀胱癌耐药细胞株

目的　用转基因法建立膀胱癌耐药细胞株 ,研究细胞耐药机制。　方法　利用脂质体(DOTAP)介导的基因转移方法 ,在浸润性膀胱癌细胞株T2 4中转入mdr1全长cDNA ,经阿霉素筛选 ,获得耐药的T2 4细胞株TADM ;免疫组化、MTT、流式细胞仪、基因组PCR、RT PCR等方法鉴定TADM的耐药表型。　结果　TADM细胞的相对耐药指数为 4 1.6 ,基因组中有mdr1cDNA插入 ,P 糖蛋白及mdr1mRNA表达增加。　结论　TADM细胞具有良好的耐药表型 ,其耐药性的产生是由mdr1全长cDNA稳定整合在T2 4细胞基因组中 ,大量表达P 糖蛋白引起。转基因法建立膀胱癌耐药细胞株具有耗时短、耐药强度高而稳定等特点。

An experimental study on the establishment of bladder cancer MDR cell line by gene transfection

Objective To establish a bladder cancer MDR cell line by gene transfection and to lay a foundation for the research of occurrence and reversion mechanisms of MDR. Methods The invasive bladder cancer T24 cells were transfected with mdr1 cDNA by cationic liposome (DOTAP) introduced gene transfection.The adriamycin (ADM) resistant cells were screened as MDR cells,named TADM. The MDR phenotype of TADM was identified by MTT, immunohistochemistry,immunofluorescence,flow cytometry,PCR and RT PCR. Results The relative resistant index of TADM was 41.6,mdr1 cDNA being integrated into the genome of T24.As a result,the expression of P gp and mdr1 mRNA in TADM increased. Conclusions The integration of mdr1 cDNA into the T24 cells greatly increases the expression of P gp.TADM cells manifests excellent MDR phenotype.The establishment of MDR cell lines by gene tranfection has the benefits of less time consuming,stronger drug resistivity and more stable.