pTet-on-PTEN-U87MG胶质瘤细胞系的构建

Establishment of pTet-on-PTEN-U87MG glioma cell line

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摘要目的建立高诱导、低背景的pTet-on-PTEN-U87MG神经胶质瘤细胞系。方法 pTet-on质粒经脂质体介导稳定转染PTEN缺失的U87MG细胞系,G418筛选,荧光素酶活性分析,挑选阳性单克隆,建立pTet-on-U87MG细胞系;然后,pTRE-PTEN反应质粒经脂质体介导稳定转染pTet-on-U87MG细胞系,潮霉素筛选,荧光素酶活性分析,挑选阳性单克隆,构建pTet-on-PTEN-U87MG细胞系。结果成功构建pTet-on-PTEN-U87MG细胞系。结论 pTet-on-PTEN-U87MG神经胶质瘤细胞系的建立,为进一步研究神经胶质瘤发病相关因素及PTEN相关的信号传导机制,提供较为理想的细胞模型。 Objective To establish pTet-on-PTEN-U87MG glioma cell line with higher induction and lower background.Methods Firstly,pTet-on-U87MG cell line was established by liposome-mediated stable transfection of PTEN-deleted U87MG cell line with pTet-on plasmid,U87MG cell line undergone G418 selection and luciferase activity analysis,and positive cell monoclone of pTet-on-U87MG was confirmed.Secondly,pTet-on-PTEN-U87MG cell line was established by liposome-mediated stable transfection of pTet-on-U87MG cell line with pTRE-PTEN responsive plasmid,pTet-on-U87MG cell line undergone hygromycine selection and luciferase activity analysis,and positive cell monoclone of pTet-on-U87MG was confirmed.Results pTet-on-PTEN-U87MG glioma cell line was successfully established.Conclusion Establishment of pTet-on-PTEN-U87MG glioma cell line makes the basis for the research of glioma pathogenesis and PTEN signal transduction with suitable cell model.

作者黄安炀程光章翔刘新平

机构地区重庆市第九人民医院神经外科第四军医大学西京医院神经外科第四军医大学生物化学教研室

出处《重庆医学》 CAS CSCD 北大核心 2012年第14期1402-1403,1406,共3页 Chongqing medicine

关键词神经胶质瘤细胞系基因报告磷酸单酯水解酶类 glioma cell line genes reporter phosphoric monoester hydrolases

分类号 R739.4 [医药卫生—肿瘤]

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