腐胺对小鼠成纤维细胞中Atp8a1基因表达的影响  

Effects of putrescine on Atp8a1 gene expression in mouse NTH3T3 cells

作　　者：万涛[1] 李朝幸[2] 田园园[2] 王李英[2] 秦栋栋[2] 李凯[2] 曲嘉琳[2] 汤华[2] 

机构地区：[1]重庆医科大学附属第一医院呼吸科,重庆400016 [2]重庆医科大学感染性疾病分子生物学教育部重点实验室,重庆400016

出　　处：《重庆医科大学学报》2011年第2期133-135,共3页Journal of Chongqing Medical University

基　　金：国家自然科学基金(编号:30671080)

摘　　要：目的:研究腐胺对小鼠成纤维细胞中Atp8a1基因表达的影响,初步探讨其作用机制。方法:用基因芯片和Real-timePCR检测正常小鼠和Azin1(Antizyme inhibitor 1)基因敲除小鼠肝脏组织中Atp8a1基因的在mRNA水平上的表达;构建Atp8a1基因启动子的虫荧光素酶报告质粒pGL3-Atp8a1-P;将重组质粒转染NTH3T3成纤维细胞中,分别在含有4μg/ml腐胺和不含腐胺的条件下,用双荧光素酶检测系统检测虫荧光素酶活性。结果:在含有腐胺的培养条件下,转染Atp8a1基因启动子虫荧光素酶报告质粒的细胞虫荧光素酶的活性明显改变。结论:腐胺能够抑制Atp8a1基因启动子活性而降低其在NIH3T3成纤维细胞中的表达。Objective： To investigate the effects of putrescine on the expression of Atp8a1 in NIH3T3 cells and reveal its regulatory mechanism.Methods： DNA microarray and Real-time PCR were performed to detect the Atp8a1 gene expression in normal mouse liver and Azin1 knockout mouse liver.Atp8a1 promoter luciferase reporter plasmid,pGL3-Atp8a1-P,was constructed.NTH3T3 cells were transiently transfected with pGL3-Atp8a1-P,and cultured in medium with and without putrescine respectively.Then the luciferase activity was detected.Results： The relative luciferase activity was changed in the cells which were transfected with pGL3-Atp8a1-P when cultured in medium added with putrescine.Conclusion： Putrescine could down-regulate the Atp8a1 gene expression in NIH3T3 cells by inhibiting its promoter＇s activity.

关 键 词：腐胺 Atp8a1 启动子 基因表达 Azin1基因敲除小鼠 

分 类 号：Q553[生物学—生物化学]