乳腺癌细胞株MCF-7-PC-1-46的建立  

Establishment of a breast cancer cell line MCF-7-PC-1-46

作　　者：常晓彤[1] 万里川[1] 张浩[1] 王健[1] 李杰之[1] 周建光[1] 黄翠芬[1] 

机构地区：[1]军事医学科学院生物工程研究所

出　　处：《生物技术通讯》2003年第1期36-38,63,共4页Letters in Biotechnology

基　　金：国家自然科学基金资助项目(30070296)

摘　　要：以人前列腺癌C4-2细胞基因组DNA为模板,扩增出PC-1基因N端编码46个氨基酸残基及其上游非编码区共599bp的DNA序列,将其正向克隆到真核表达载体pIRES2中,并在脂质体介导下,转染人乳腺癌细胞MCF-7,经G418筛选获得阳性单克隆,细胞扩大培养后,进行PCR和RT-PCR分析,检测外源PC-1基因在靶细胞中的整合与转录。PCR和RT-PCR结果表明,稳定转染细胞株MCF-7-PC-1-46具有外源目的基因的整合和相应mRNA的高表达。说明成功建立了稳定表达外源PC-1基因N端46个氨基酸的人乳腺癌细胞株,为进一步研究PC-1基因的生物学功能提供了实验材料。The eukaryotic expression vector pIRES2-PC-1-46was construc te d by inserting a599bp DNA fragment ,which consisted of the N-terminus46am ino acids sequence and its upstream untranslation region of human PC-1gene,in to the vector pIRES2.The human breast cancer cell MCF-7was transfected with pIRES2-PC-1-46using Lipofectamine regent.The positive cell clones were s elected with G418.Utilizing PCR and RT-PCR technologies,the insertion and expression of exogenous PC-1gene in MCF-7cells were analysed.The results of stable transfec-tion and high expression of exogenous PC-1gene in MCF-7cel ls were confirmed by PCR and RT-PCR.The breast cancer cell line MCF-7-PC-1-46that stably expressed PC-1gene N-terminus46amino acids was establis hed.It will facilitate further studies on biological function of the PC-1gene .

关 键 词：乳腺癌细胞株 MCF-7-PC-1-46 PC-1基因 稳定转染 基因表达 

分 类 号：R737.9[医药卫生—肿瘤]