mTOR基因转染对NIH3T3成纤维细胞增生的影响  

Effects of mTOR Transfection on Proliferation of NIH3T3 Fibroblasts

作　　者：李小明[1] 杨作成[1] 陈淳媛[1] 李卓颖[1] 

机构地区：[1]中南大学湘雅三医院儿科,长沙410013

出　　处：《医学研究杂志》2008年第3期32-34,共3页Journal of Medical Research

基　　金：湖南省卫生厅科研基金资助(B2005-072);教育部留学回归启动基金资助

摘　　要：目的探讨mTOR基因转染对NIH3T3成纤维细胞增生的影响。方法运用电穿孔法将pcDNA3-mTOR质粒转染NIH3T3成纤维细胞,经G418筛选抗性克隆,RT-PCR和Western印迹分析检测转染组和对照组细胞mTOR mRNA与蛋白的表达,MTT方法检测细胞增生。结果转染组细胞mTOR mRNA和蛋白质的表达显著增强(P<0.01);MTT法检测转染组光吸收值显著大于对照组(P<0.01);结论mTOR基因转染成纤维细胞可获稳定表达,并明显促进成纤维细胞增生。Objective To study the effects of mammalian target of rapamycin （mTOR） transfection on the proliferation of NIH3T3 fibroblasts. Methods The plasmid of pcDNA3 - roTOR was transfected into NIH3T3 fibroblasts with electroporation method. The positive cell clones were selected with G418. The stable mRNA and protein expressions of mTOR in the cells were determined by RT- PCR and Western blotting analysis respectively. MTT assay was employed to observe the proliferation of NIH3T3 fibroblasts. Results mTOR mRNA and protein expressions increased obviously in transfected group than that of in control group（ P 〈 0.01 ）, MTT assay showed the Aborption values in transfected group rose significantly than that in control group （ P 〈 0.01 ）. Conclusions The stable expression of mTOR gene in NIH3T3 fibroblasts transfected with pcDNA3 - mTOR could be obtained, mTOR gene transfection can stimulate the proliferation of NIH3T3 fibroblasts.

关 键 词：MTOR 基因转染 成纤维细胞 细胞增生 

分 类 号：R364.3[医药卫生—病理学] R977[医药卫生—基础医学]