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| 利用Tet-on调控系统建立人肝癌HepG2Tet-on细胞系 | |
| 引用本文： | 金炜东,陈孝平,陈钢.利用Tet-on调控系统建立人肝癌HepG2Tet-on细胞系[J].中国普通外科杂志,2009,18(2):10-149. |
| 作者姓名： | 金炜东 陈孝平 陈钢 |
| 作者单位： | 1. 广州军区武汉总医院,普通外科,湖北,武汉,430070 2. 华中科技大学同济医学院附属同济医院,肝脏外科,湖北,武汉,430030 3. 温州医学院附属第一医院,肝胆外科,浙江,温州,325000 |
| 摘 要： | 目的 构建可以用强力霉素调控表达的人肝癌HepG2Tet-on细胞系,为进一步研究肝癌相关基因功能奠定基础.方法 用脂质体转染法将pWHE146质粒转染到人肝癌HepG2细胞中,用G418筛选出稳定表达细胞克隆;单克隆分别扩增后,瞬时转染pTRE-hyg-luc质粒;强力霉素诱导表达后,检测荧光素酶表达活性,挑选出受强力霉素调控的低背景、高表达的HepG2Tet-on细胞株.结果 成功构建了一株受强力霉素调控的高表达低背景的HepG2Tet-on细胞株(诱导倍数达154.106倍).结论 HepG2Tet-on细胞株可用于外源基因的真核调控高表达,为研究真核基因功能提供一种可靠的细胞株. |
| 关 键 词： | 肝肿瘤 转染 诱导 基因表达 Tet-on基因表达调控系统 |
| 收稿时间： | 1900/1/1 0:00:00 |
| 修稿时间： | 1900/1/1 0:00:00 |
Establishment of HepG2Tet-on cell line controlled by the Tet-on regulatory system |
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| JIN Weidong,CHEN Xiaoping,CHEN Gang.Establishment of HepG2Tet-on cell line controlled by the Tet-on regulatory system[J].Chinese Journal of General Surgery,2009,18(2):10-149. | |
| Authors: | JIN Weidong CHEN Xiaoping CHEN Gang |
| Institution: | （1.Department of General Surgery, Wuhan General Hospital of Guangzhou Command, Wuhan 430070, China; 2.Department of Hepatic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China; 3.Department of Hepatobiliary Surgery, the First Affiliated Hospital, Wenzhou Medical College， Wenzhou,Zhejiang 325000,China） |
| Abstract: | Objective:To establish tetracycline-controlled inducible system (Tet-on) in HepG2 cell for further research of the function of related gens.Methods :The HepG2 cells were transfected with pWHE146 vector by using liposome transfection reagent. The transfected cells were selected in medium containing G418, and G418-resistant clones were isolated. All individual G418-resistant clones were screened by transient transfection with plasmid pTRE-hyg-luc for clones with low background and high induction of luciferase in response to Dox. Results:One HepG2 cell line, which exhibited high levels of induction（154,106 times）and high gene expression levels, was obtained.Conclusions:The HepG2 cell line can be used to highly express eukaryotic gene and this Tet-on system is available for use in eukaryotic gene function studies. |
| Keywords: | Liver Neoplasms Transfection Inducible Gene Expression Tet-on System |
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