乳腺癌耐药蛋白(BCRP)高表达耐药细胞系的建立及其耐药机制的初步研究.pdf

．768． 乳腺癌耐药蛋白(BCRP)高表达耐药细胞系的 建立及其耐药机制的初步研究：l。 cancerresistance 摘要 目的：建立稳定表达乳腺癌耐药蛋白(Breast protein，BCRP)的小鼠成纤维细胞系 导入真核表达载体pcDNA3．1，转化感受态E．Coli RT-PcR检测BCRP的mRNA表达，Westernblot检测BCRP蛋白表达。MrrI法检测耐药克隆PA317／BCRP细胞对米 blot检测 RT-PCR和Westemblot，均检测到细胞中BCRP基因转录及蛋白表达。与空质粒转染细胞、未转染细胞对照组相 的表达，观察到耐药细胞的AKT表达含量明显高于转染空质粒和未转染的PA317细胞。推测BCRP可能通过影响 AKT表达上调发挥其耐药作用。 关键词 乳腺癌耐药蛋白，转染PA317细胞耐药信号转导 doi：10．39690．issn．1000-8179．2009．13．014 Researchon EstablishmentofCeULinewith ofBCRPandBasic HighExpression Mechanism Drug—-resistant ZHANG Huilai。SUNYan，SONGZheng，LlUXianming，WANGHuaqing author：WANG 63．com Coffesponding Huaqing，E—mail：huaqingw@1 of Instituteand of Medical 300060，China DepartmentLymphoma，CancerHospitalTianjin University,Tianjin Grant NaturaIScienceFoundationof support：NationaI China acelIline themechanismofits Abstract establish BCRPand 0bjective：To expressing investigate drug resistarice．Methods：Theaimed wereisolatedfromthecelIIineMcf-7，ADRreverse segments by transcription chainreactJonandwerejnsertedjntoa toconstructa polymerase eukaryoticexpressionplasmidpcDNA3．1 first inEsche- recombinant recombinantwas expressionplasmidpcDNA3．I／BCRP．Theplasmid propagated richiacoliDH5a．thenwasextractedand and wi