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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。
Objective To establish a bladder cancer resistant cell line by transgenic method and study the mechanism of cell resistance. Methods The full-length cDNA of mdr1 was transfected into invasive bladder cancer cell line T2 4 by lipofectamine (DOTAP) -mediated gene transfer. The adherent cells were screened by doxorubicin to obtain TADM resistant T2 4 cell line. Immunohistochemistry MTT, flow cytometry, genomic PCR, RT PCR and other methods to identify the resistant phenotype of TADM. Results The relative resistance index of TADM cells was 41.6. The mdr1 cDNA was inserted into the genome and the expressions of P-glycoprotein and mdr1 mRNA were increased. Conclusion TADM cells have a good resistance phenotype. The resistance of TADM cells is caused by the stable integration of mdr1 full-length cDNA in the genome of T2 4 cells and the massive expression of P-glycoprotein. Transgenic method to establish bladder cancer drug resistant cell lines with short time-consuming, high drug resistance and stability and so on.