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.
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