基于H9C2细胞株的ADRB1基因多态性稳定过表达模型的构建  

Establishment of stable over-expression model of ADRB1gene polymorphism based on H9C2 cell line

作　　者：陈丽萍 肖婷 邓懋清 谢珊珊 吴定昌 Chen Liping;Xiao Ting;Deng Maoqing;Xie Shanshan;Wu Dingchang(Department of Clinical Laboratory,Longyan First Hospital,Longyan First Affiliated Hospital of Fujian Medical University,Longyan 364000,Fujian,China)

机构地区：[1]福建省龙岩市第一医院,福建医科大学附属龙岩第一医院检验科,福建龙岩364000

出　　处：《右江民族医学院学报》2022年第2期180-183,194,共5页Journal of Youjiang Medical University for Nationalities

基　　金：福建省自然科学基金项目(2018J01403);福建省医学创新课题(2016-CX-49)。

摘　　要：目的构建β1肾上腺素能受体(ADRB1,1165G>C)基因多态性稳定过表达的心肌细胞模型,为其在高血压、心力衰竭及不良心血管事件中的功能及机制研究奠定了实验基础。方法将ADRB1cDNA克隆片段和PMT399载体连接后获得pSE4077和pSE4078重组质粒,再与慢病毒包装质粒共转染293T细胞,收集上清液感染H9C2细胞,利用定量聚合酶链式反应(QPCR)方法筛选稳转细胞株。结果pSE4077和pSE4078组的ADRB1蛋白表达水平显著高于未转染的细胞对照组和转染空病毒的对照组,且pSE4078组也高于pSE4077组。结论基于H9C2细胞株的ADRB1基多态性稳定过表达模型构建成功。Objective To establish a cardiomyocyte model with stable over-expression ofβ1adrenergic receptor(ADRB1,1165G>C)gene polymorphism,so as to lay an experimental foundation for the study of its function and mechanism in hypertension,heart failure and other adverse cardiovascular events.Methods After the ligation of ADRB1cDNA clone fragments and PMT399vector,the recombinant plasmids pSE4077and pSE4078were co-transfected with lentivirus package plasmid into 293Tcells.The supernatant was collected to infect H9C2cells.Stably transfected cells were screened by quantitative polymerase chain reaction(QPCR).Results The expressions of ADRB1in pSE4077and pSE4078groups were significantly higher than those in the control group of untransfected cells and those in the control group transfected with empty virus.And those of the pSE4078group was also higher than those of the pSE4077group.Conclusion The stable over-expression models of ADRB1gene polymorphism have been successfully constructed based on H9C2cell line.

关 键 词：高血压 ADRB1 H9C2细胞株 慢病毒载体 基因多态性 

分 类 号：R544.1[医药卫生—心血管疾病]