MCF-7/BCRP细胞系的建立及其生物学特征  

Establishment of breast cancer resistance protein expression cell line MCF-7/BCRP and its biological characterization

作　　者：李文通[1] 周庚寅[1] 宋现让[2] 迟伟玲[3] 任瑞美[2] 王兴武[2] 

机构地区：[1]山东大学医学院病理学教研室,山东济南250012 [2]山东省肿瘤医院基础研究中心,山东济南250117 [3]山东大学医学院分子生物学教研室,山东济南250012

出　　处：《山东大学学报（医学版）》2005年第6期469-472,共4页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金资助课题(30300124);高等学校博士学科点专项基金资助课题(20020422042)。

摘　　要：目的:建立表达乳腺癌耐药蛋白(BCRP)的耐药细胞系MCF-7/BCRP。方法:提取MCF-7细胞的总RNA,设计BCRP基因上下游引物,用RT-PCR法克隆出BCRP基因全长并连接到真核表达载体pcDNA3.1穴-雪上,转染MCF-7细胞后以G418筛选细胞。采用米托蒽醌外排实验、细胞毒性实验、细胞群体倍增时间、免疫荧光法鉴定构建的耐药细胞系。结果:MCF-7/BCRP对米托蒽醌耐药指数增加至14.72;外排米托蒽醌的作用增强;细胞群体倍增时间较亲代细胞延长;MCF-7/BCRP细胞能表达一定量的BCRP。结论:MCF-7/BCRP细胞能够表达BCRP并具有BCRP的耐药表型。Objective: To construct the breast cancer resistance protein(BCRP) expression cell line MCF-7/BCRP. Methods: The total RNA of MCF-7 cells was extracted, the whole length of BCRP gene by RT-PCR was cloned and ligated to pcDNA3.1(-), the positive clones were transfected to MCF-7 cells and were selected positive cell clones with G418, the survival cells were identified by MTT, efflux assay and immunofluorescencere assay. Results: The multidrug resistance index of MCF-7/BCRP to mitoxantrone increased to 14.72; the efflux of MCF-7/BCRP to mitoxantrone enhanced; BCRP could be detected by immunofluorescence assay and it mainly located on the membrane of MCF-7/BCRP cells. Conclusion: MCF-7/BCRP is a reliable BCRP expression cell model, the establishment of this cell line has laid basis for further study of the mechanism of BCRP.

关 键 词：多药耐药相关蛋白类 乳腺肿瘤 MCF-7/BCRP细胞系 基因表达 

分 类 号：Q786[生物学—分子生物学]