论文部分内容阅读
以逆转录病毒介导，将人全长mdr-1cDNA导入人肺腺癌细胞系GLC82中，经G418和阿霉素筛选，挑选出3个阳性克隆。用mdr-1cDNA的一种特异引物在3个克隆中都扩增到1条167bp的带，表明mdr-1基因cDNA已稳定地整合在染色体基因组中。原位杂交显示转染细胞的mdr-1转录升高，免疫细胞化学发现转染细胞中P170糖蛋白呈弱阳性。MTT药敏实验表明3个克隆对阿霉素的抗药性分别是GLC细胞的6.4、7.0和8.8倍。提示在mdr-1基因cDNA转染的细胞中，mdr-1基因低表达足以大大提高细胞对药物的耐受性。
The human full-length mdr-1 cDNA was introduced into human lung adenocarcinoma cell line GLC82 by retrovirus-mediated selection. Three positive clones were selected by G418 and adriamycin screening. Using a specific primer for mdr-1 cDNA, a 167 bp band was amplified in all three clones, indicating that the cDNA of mdr-1 gene has been stably integrated into the chromosomal genome. In situ hybridization showed that the mdr-1 transcription of the transfected cells was increased. Immunocytochemistry revealed that the P170 glycoprotein was weakly positive in the transfected cells. MTT susceptibility testing showed that the resistance of three clones to adriamycin was 6.4, 7.0 and 8.8 times that of GLC cells, respectively. This suggests that the low expression of the mdr-1 gene in cells transfected with the mdr-1 gene cDNA is sufficient to significantly increase the drug resistance of the cells.