稳定表达核仁素的H9C2细胞株的建立和鉴定  

Establishment of H9C2 Cell Line Stably Expressing Nucleolin

作　　者：宋娟[1,2] 张任飞[3] 张彬[1] 蒋碧梅[1] 

机构地区：[1]中南大学湘雅医学院,在读研究生410078 [2]长治医学院病理生理学教研室 [3]长治医学院附属和平医院

出　　处：《长治医学院学报》2009年第2期88-90,共3页Journal of Changzhi Medical College

基　　金：湖南省科技计划资助(2007JT3022)

摘　　要：目的:建立稳定表达核仁素的H9C2大鼠心肌细胞株,为进一步研究核仁素在心肌细胞凋亡中的作用提供细胞模型。方法:将含有核仁素全长的质粒pCMV-Tag2B-Nuc用脂质体转染技术导入H9C2细胞,使核仁素在细胞内表达,用G418筛选出稳定表达核仁素的细胞株;用双酶切、DNA测序鉴定质粒,用RT-PCR及Western blot检测核仁素基因在靶细胞中的整合与表达。结果:经G418反复筛选的H9C2细胞株中,核仁素基因在mRNA及蛋白质水平表达均较野生型细胞及转空载体细胞升高。结论:用含核仁素全长的质粒转染H9C2大鼠心肌细胞,经筛选后可稳定表达核仁素。Objective：To establish H9C2 cell line that can stably express the nucleolin,which can provide a cell line with which we can analyze the function of nucleolin in apoptosis of cardiomyocytes. Methods：The pCMV - Tag2B - Nuc plasmid was transfeeted into H9C2 cells with lipofectin,the stable transfectants screened by G418 and the expression of nucleolin was identified and analyzed by RT - PCR and Western blot. The pCMV - Tag2B - Nuc plasmid was confirmed by double - enzyme digestion and sequencing. Resuits：After G418 screening, compared with the control group, the mRNA and protein of nucleolin in transfected cell line were highly expressed, as demonstrated by the RT - PCR and Western blot analysis. Conclusion ： The rat H9C2 cell line stably expressing nucleolin was successfully constructed.

关 键 词：核仁素 H9C2细胞 基因转染 

分 类 号：R363[医药卫生—病理学]