Zhou Z, Lin C, Chen F, Luo X, Wei H
Cancer Institute, CAMS, Beijing.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1997 Feb;19(1):67-71.
Human lung cancer leads the mortality of cancers and the chemotherapy is often uneffective because of drug resistance. In order to study the role of mdr-1 gene in resistant lung cancer, the fully length mdr-1 cDNA was transferred into a sensitive lung cancer cell line GLC. The mdr-1 cDNA was constructed in a retroviral vector, pDORneo. The transfection of recombinant plasmid was carried out by lipofectin. Supernatant containing infective viruses derived from a G418 resistant clone of package cell PA317 was used to infect GLC cell which is sensitive to chemotherapeutic agents. After G418 and adriamycine selections, three P-glycoprotein positive clones were isolated and the integration of mdr-1 cDNA was demonstrated by PCR of genomic DNA. The relative resistance of 3 clones to adriamycine as and elevated by 5.4, 6.0 respectively 7.8 times compared with the untransfected cell and the transcription of mdr-1 gene in these transfected cells as obviously enhanced by in situ hybridization. This results suggest that the mdr-1 gene plays a role in increasing drug resistance of human lung cancer.
人类肺癌导致癌症死亡率居高不下，且由于耐药性，化疗往往无效。为了研究多药耐药基因1（mdr-1）在耐药肺癌中的作用，将全长mdr-1 cDNA转入敏感肺癌细胞系GLC。mdr-1 cDNA构建于逆转录病毒载体pDORneo中。重组质粒的转染通过脂质体转染法进行。含有源自包装细胞PA317的G418抗性克隆的感染性病毒的上清液用于感染对化疗药物敏感的GLC细胞。经过G418和阿霉素筛选，分离出三个P-糖蛋白阳性克隆，并通过基因组DNA的PCR证明了mdr-1 cDNA的整合。与未转染细胞相比，3个克隆对阿霉素的相对耐药性分别提高了5.4倍、6.0倍和7.8倍，并且通过原位杂交明显增强了这些转染细胞中mdr-1基因的转录。这些结果表明mdr-1基因在增加人类肺癌耐药性中起作用。
