逆转录病毒转染法建立耐药性人膀胱癌细胞系

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ｍｄｒ１ ｍＲＮＡ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２１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

MDR HUMAN BLADDER CANCER CELLS LINE ESTABLISHED BY RETROVIRUS INFECTION

Objective To establish a MDR human bladder cancer cells line.Methods Human bladder cancer cells were transfected by mdr1 retrovirus supernatant. Multidrug resistance of cancer cells was tested by MTT assay; expression of P GP on membrane and mdr1 mRNA in cells was detected with immunocytochemistry and in situ hybridization. The presence of mdr1 cDNA in genome of pT24mdr1 was detected by polymerase chain reaction (PCR). Results The pT24mdr1 cell line was found to be 8 times more resistant to adriamycin and 21 times to colchicine than T24, and showed stable resistance to both adriamycin and colchicine; through immunocytochemistry assay with McAb against P GP, pT24mdr1 cells expressed P GP higher than T24 on the membrane. The pT24mdr1 cells showed much higher mdr1 mRNA level in contrast to T24 by in situ hybridization. The result of polymerase chain reaction verified the presence of mdr1 cDNA in genome of pT24mdr1.Conclusion Gene transfer by retrovirus is a satisfied method in establishing MDR human bladder cancer cells line. $$$$