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肝癌先天性多表达多药耐药基因,严重影响肝癌的化疗效果,筛选肝癌细胞中的耐药基因,研究其耐药机制有助于提高肝癌化疗效果,提高肝癌的治愈率。首先构建肝癌细胞逆转录病毒的cDNA文库,感染成纤维细胞,使得逆转录病毒基因整合进细胞,加药筛选,存活细胞中的基因再次包装成病毒,用于下一轮筛选。采用循环包装回收(Cyclical packaging rescue,CPR)技术进行肝癌细胞耐药基因的筛选即是通过病毒包装将基因从细胞中钓取出来,相比于常规筛选方法,仅通过一轮筛选可能会出现很多假阳性基因,采用CPR技术则是通过多轮筛选,很大程度减少了假阳性细胞的出现。通过该方法经过四轮筛选获得核糖体蛋白S11(RPS11)、核糖体蛋白L6(RPL6)、核糖体蛋白L11(RPL11)、核糖体蛋白L24(RPL24)等几种基因,经初步检测,增加了细胞的耐药性。
Liver cancer congenital multi-drug resistance gene expression, serious impact on the efficacy of chemotherapy of liver cancer, screening of drug-resistant genes in liver cancer cells, study of its drug resistance mechanisms can help improve the efficacy of liver cancer chemotherapy and improve the cure rate of liver cancer. First of all, a cDNA library of retrovirus for liver cancer cells was constructed to infect the fibroblasts so that the retrovirus genes were integrated into the cells. Drug selection and screening, surviving cells were repackaged into viruses for further screening. Cyclical packaging rescue (CPR) technology for screening drug-resistant genes in hepatocellular carcinoma cells is a method of obtaining genes from cells by virus packaging. Compared with conventional screening methods, a lot of screening by one round of screening may occur False positive genes, using CPR technology is through multiple rounds of screening, greatly reducing the appearance of false positive cells. After four rounds of screening, several genes of ribosomal protein S11 (RPS11), ribosomal protein L6 (RPL6), ribosomal protein L11 (RPL11) and ribosomal protein L24 (RPL24) Cell resistance.