【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。更多还原

【Abstract】 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.更多还原