肝癌多药耐药基因工程细胞的建立及特性分析  

Analysis of biological characteristics for the hepatocellular carcinoma engineering cell lines with multidrug resistance

作　　者：李波[1] 夏洪芬[2] 李德华 陈永兵[4] 苏松[1] 张孟瑜[1] 贺凯[1] 冯春红[1] 徐松波[1] 倪建彬[1] 罗亮[1] 夏先明[1] 

机构地区：[1]泸州医学院附属第一医院肝胆外科,四川泸州646000 [2]泸州医学院附属第一医院手术室,四川泸州646000 [3]成都地奥制药集团有限公司基因工程药物研究室,四川成都610041 [4]首都医科大学附属佑安医院肝胆外科,北京100069

出　　处：《中国现代医学杂志》2009年第14期2123-2126,共4页China Journal of Modern Medicine

基　　金：泸州医学院和四川省科技厅科研资助项目(No:05JY029-146)

摘　　要：目的利用转基因方法建立表达mdr1基因的肝癌基因工程细胞,为深入肝癌多药耐药现象研究提供理想的细胞模型。方法构建表达mdr1cDNA全长序列的真核表达载体PCI/mdr1,利用脂质体将mdr1cDNA转染入人肝癌细胞株HepG2细胞,通过阿霉素短暂诱导和G418筛选转染阳性细胞,筛选出稳定表达mdr1及P-gp蛋白的细胞株HepG2/R。结果HepG2/R细胞与出发株HepG2细胞相比较,MTT法检测生长曲线并无明显差异,对阿霉素和丝裂霉素的耐药性分别增加了35.7倍和125倍,RT-PCR检测HepG2/R细胞的mdr1mRNA表达明显增加,流式细胞仪检测HepG2/R细胞P-gp的表达状况明显增加。结论成功建立表达mdr1基因的肝癌工程细胞株。[Objective] To construct human hepatocenular genetic engineering ceils expressing mdr1 gene for the experiment of studying the muhidrug resistance in hepatocellular carcinoma. [Methods] The mammalian expression vector pCI/mdr1 expressing the whole human mdr1 eDNA sequences has been constructed. The plasmid pCI/mdr1 was transferred to human hepatocellular carcinoma cell line HepG2 by liposome. The engineering cell line HepG2/R only stably expressing mdr1 gene and p-glycoprotion was developed by transient adriamycin inducement and G418 filtration. [Results] Compared with the parental HepG2 cells, the growth and proliferation of HepG2/R cells was not affected by the mdr1 transfer, the sensitivity of cells to adriamycin and mitomyein was increased by 35 times and 125 times respectively by MTT assay, the levels of mdr1 mRNA and p-glycoprotion expression were upregulated obviously by RT-PCR and flow cytometry respectively. [Conclusions] HepG2/R cells transfected with pCI/mdr1 can efficiently express mdr1 gent, which is potentially applicable to multidrug resistance experiment of hepatocellular carcinoma.

关 键 词：多药耐药性 多药耐药基因 肝癌细胞株 工程细胞株 

分 类 号：Q813[生物学—生物工程] R735.7[医药卫生—肿瘤]