ABCG2食管癌耐药细胞系Eca109/ABCG2的建立  

Establishment of Eca109/ABCG_2,a drug resistant ABCG_2 esophageal cancer cell line

作　　者：刘亮[1] 左静[2] 赵丽[1] 王静[1] 郭建文[1] 刘江惠[1] 左连富[1] 

机构地区：[1]河北医科大学第四医院肿瘤研究所,石家庄050011 [2]河北医科大学第四医院肿瘤内科,石家庄050011

出　　处：《解放军医学杂志》2010年第1期32-35,共4页Medical Journal of Chinese People's Liberation Army

基　　金：河北省自然科学基金(C2007001060);河北省普通高等学校强势特色学科肿瘤学建设经费资助课题(冀教高[2005]52号)

摘　　要：目的探讨腺苷三磷酸结合转运蛋白G超家族成员2(ABCG2)在介导食管癌多药耐药中的作用。方法扩增ABCG2基因,与pcDNA3.1载体连接,构建pcDNA3.1-ABCG2重组质粒,转染Eca109细胞后用G418筛选稳定转染细胞。采用流式细胞术检测细胞内阿霉素的含量,评价细胞对药物的外排作用。MTT法检测阿霉素对Eca109/ABCG2、Eca109/pcDNA3.1、Eca109细胞生长的抑制作用,并计算半数抑制浓度(IC50)和细胞的耐药指数。采用Western blotting和RT-PCR法检测细胞中ABCG2蛋白及mRNA的表达量。结果成功建立转染ABCG2基因的Eca109/ABCG2细胞。Eca109/ABCG2细胞中ABCG2的mRNA和蛋白表达量升高。阿霉素对Eca109/ABCG2、Eca109/pcDNA3.1、Eca109细胞的IC50值分别为18.61±3.94、4.18±0.14、4.69±0.88μg/ml。Eca109/AB-CG2细胞对阿霉素的耐药指数为3.97,且外排阿霉素的作用增强。结论Eca109/ABCG2细胞具有ABCG2的耐药表型,能够稳定表达ABCG2蛋白,可作为研究ABCG2介导的多药耐药相关机制的细胞模型。Objective To explore the role of ATP binding cassette G2（ABCG2）in mediating multidrug resistance（MDR）in esophageal cancer.Methods The recombinant plasmid pcDNA3.1-ABCG2 was constructed by amplifing ABCG2 gene and then conjugating with pcDNA3.1 vector.The plasmid was then transfected into Eca109 cells,and positive cell clones were selected with G418.The concentration of adriamycin（ADM）within cells were examined by flow cytometry to evaluate the eliminating effects of cells.The inhibition effects of ADM on the growth of Eca109/ABCG2,Eca109/pcDNA3.1 and Eca109 cells were detected by MTT assay,and the 50% inhibitory concentration（IC50）of ADM and resistence index of cells were calculated.The expressions of protein and mRNA of ABCG2 were detected by Western blotting and RT-PCR.Results The Eca109/ABCG2 cells which were transfected with ABCG2 gene were successfully established.The up-regulated expressions of ABCG2 protein and mRNA were detected in Eca109/ABCG2 cells.The IC50 values of ADM against Eca109/ABCG2,Eca109/pcDNA3.1 and Eca109 cell were 18.61±3.94,4.18±0.14 and 4.69±0.88,respectively.Moreover,Eca109/ABCG2,with a resistance index of 3.97 to ADM,showed a stronger eliminating effect on ADM than the other type of cells.Conclusion Eca109/ABCG2 cell line,with a typical phenotype of ABCG2,may stably express ABCG2 protein,and may be an ideal cell model for further research on drug resistance.

关 键 词：三磷酸腺苷结合转运蛋白G超家族成员2 多药耐药相关蛋白质类 食管肿瘤 

分 类 号：R735.1[医药卫生—肿瘤]