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作 者:刘立[1] 彭健[1] 李静琪[1] 吕立夏[1] 李学礼[1] 徐磊[1]
机构地区:[1]同济大学医学院生物化学教研室,上海200092
出 处:《同济大学学报(医学版)》2007年第5期43-47,共5页Journal of Tongji University(Medical Science)
摘 要:目的探讨TMEFF2基因过表达对体外培养SH-SY5Y细胞增殖的影响。方法克隆全长的TMEFF2基因,连接入pEGFP-N2载体。以脂质体包裹重组质粒转染SH-SY5Y细胞。在细胞中可表达生成TMEFF2-EGFP融合蛋白。经G418筛选,建立TMEFF2过表达细胞株,同时筛选作为对照的pEGFP-N2稳定转染细胞株。MTT法测定TMEFF2过表达细胞株和pEGFP-N2稳定转染细胞株以及SH-SY5Y细胞增殖。结果成功构建真核表达质粒pEGFP-TMEFF2,并筛选得到稳定表达细胞株。在细胞接种密度较高时,TMEFF2过表达细胞株的增殖较pEGFP-N2稳定转染细胞株以及SH-SY5Y细胞株均明显降低。结论TMEFF2基因可以抑制SH-SY5Y细胞的增殖,且抑制作用具有明显密度依赖性。Objective To investigate the effects of TMEFF2 gene on proliferation of SH-SY5Y cells. Methods Plasmid pEGFP-TMEFF2 was constructed and transfected into SH-SY5Y cells by using Lipofectamine, Cells stably expressing TMEFF2 were selected with G418. Cells stably transfected with pEGFP-N2 were selected with same method. Proliferation of pEGFP-TMEFF2 transfected cells, pEGFP-N2 transfected cells and SH- SY5Y cells were evaluated with MTT. Results Cells stably transfected with pEGFP-TMEFF2 and cells stably transfected with pEGFP-N2 were constructed successfully, When inoculated with high density, the proliferation activity of pEGFP-TMEFF2 transfected cells decreased more than pEGFP-N2 transfected cells and SH-SY5Y cells. Conclusion TMEFF2 significantly inhibits the proliferation of SH-SY5Y cells, and the inhibition is density dependent.
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