人TCF21基因真核表达载体的构建及稳定转染肺腺癌顺铂耐药细胞系A549/DDP的建立  

Construction of eukaryotic expression vector of human TCF21 and establishment of stably transfected human lung adenocarcinoma cisplatin-resistant cell line A549/DDP

作　　者：邬成业[1] 轩伟霞[2] 杨会珍 张群成 李玉光[2] WU Cheng-ye;XUAN Wei-xia;YANG Hui-zhen;ZHANG Qun-cheng;LI Yu-guang(Institute of Hematology,Henan Province People ＂s Hospital,Zhengzhou,Henan 450003,Chin)

机构地区：[1]河南省人民医院血液病研究所,河南郑州450003 [2]河南省人民医院呼吸内科,河南郑州450003

出　　处：《现代预防医学》2018年第14期2610-2614,共5页Modern Preventive Medicine

基　　金：河南省科技攻关项目(152102310413)TCF21逆转人肺腺癌细胞耐药的作用及其机制的研究

摘　　要：目的建立稳定高表达人转录因子21(TCF21)基因的肺腺癌顺铂耐药细胞系(A549/DDP)。方法从人肝癌细胞Hep G2中提取总RNA,应用RT-PCR技术获得人TCF21基因的c DNA,并构建p EGFP-N1-TCF21真核表达载体;用脂质体转染A549/DDP细胞,经G418筛选获得稳定表达人TCF21基因的细胞系;运用RT-PCR、Western-Blot技术对细胞系进行鉴定。结果成功构建了人TCF21基因真核表达载体,建立了能够稳定高效表达人TCF21基因的A549/DDP细胞系。结论 TCF21基因真核表达载体的构建及稳定转染A549/DDP细胞系的建立为探究其逆转人肺腺癌细胞耐药的作用及其分子机制奠定了坚实的实验基础。Objective To establish the human lung adenocarcinoma cisplatin-resistant cell line（A549/DDP）that stably over expresses transcription Factor 21（TCF21）. Methods The total RNA was extracted from human Hep G2, and TCF21 c DNAs were cloned by RT-PCR, then they were inserted into the eukaryotic expression vectors p EGFP-N1. The A549/DDP cell line was stably transfected with p EGFP-N1-TCF21 plasmids by Lipofectamine2000, then the transfected cells were selected by G418（400 μg·m L-1）. The expression of TCF21 on transfected cells was verified through RT-PCR, Western-Blot. Results The eukaryotic expression vectors were successfully constructed and the A549/DDP cell line with stable TCF21 over expression was obtained. Conclusion The construction of the eukaryotic expression vector p EGFP-N1-TCF21 and the establishment of stable A549/DDP cell line with TCF21 expression provide a strong experimental basis for study its role and molecular mechanism in reversing human lung adenocarcinoma cell resistance.

关 键 词：TCF21基因 转染 A549/DDP细胞 

分 类 号：R181.24[医药卫生—流行病学]