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目的 建立高效、稳定的人膀胱癌多药耐药性细胞模型。方法 应用逆转录病毒转染法建立人膀胱癌耐药性细胞模型 ;MTT法检测细胞系在不同化疗药物作用下的生存率 ,免疫组化检测肿瘤的 P- GP表达 ,原位杂交检测肿瘤细胞 mdr1m RNA的表达量 ,PCR检测 mdr1基因转移至细胞基因组片段。结果 转基因细胞系对阿霉素、秋水仙素的耐药性分别提高 8倍和 2 1倍 ,免疫组化示转基因细胞系 P-GP表达量增强 ,原位杂交结果示肿瘤细胞内 m dr1m RNA的表达量明显增加 ,PCR表明基因转移后细胞内能扩增出mdr1片段。结论 逆转录病毒转染法 ,是建立人膀胱癌多药耐药性细胞模型的一种较理想的方法。
Objective To establish an efficient and stable multidrug resistant human bladder cancer cell model. Methods The human bladder cancer drug-resistant cell model was established by retrovirus transfection method. The survival rate of the cell line was detected by MTT assay, the expression of P-GP was detected by immunohistochemistry and the tumor cells were detected by in situ hybridization mdr1m RNA expression, PCR detection of mdr1 gene transfer to the cell genome fragment. Results The resistance of the transgenic cell lines to adriamycin and colchicine increased by 8-fold and 21-fold, respectively. The expression of P-GP in the transgenic cell lines was enhanced by immunohistochemistry. The in situ hybridization results showed that the m dr1m RNA The expression of mdr1 gene was significantly increased after PCR. Conclusion Retrovirus transfection method is an ideal method to establish multidrug resistance cell model of human bladder cancer.