精胺对Azin1基因敲除小鼠体内Atp8a1基因表达调控的研究

Effects of Spermine on Atp8a1 Gene Expression in Azin1 Knockout Mice

目的 研究精胺对Azin1基因敲除小鼠体内Atp8a1基因表达的影响,并对其作用机理进行初步的探讨.方法 利用Real-time PCR检测正常小鼠和Azin1基因敲除小鼠体内Atp8a1基因的在mRNA水平上的差异表达情况;构建Atp8a1基因的启动子虫荧光素酶报告质粒;将启动子重组质粒转染NTH3T3细胞后,在细胞培养液中加入精胺,检测精胺对启动子活性的影响.结果 Atp8a1基因在Azin1基因敲除小鼠体内的表达量明显增加;精胺能够增强Atp8a1的启动子活性.结论 精胺能够通过增强Atp8a1的启动子活性而增强其在Ain1基因敲除小鼠体内转录水平的表达.

Objective To investigate the effects of spermine on the expression of Atp8al in Azinl knockout mice and reveal its regulatory mechanism. Methods Real-time PCR was performed to detect the Atp8al expression in normal mice and Azinl knockout mice. Atp8al promoter luciferase reporter plasmid, pGL3-AtpSal-P, was constructed. NTH3T3 cells were transiently cotransfected with the same amount of pGL3-AtpSal-P and pRL-TK, and then the transfected NTH3T3 cell culture medium was added with spermine. The promoter activity and the effect of spermine on the promoter activity were analyzed by dual luciferase reporter system. Results The Atp8al gene expression in Azinl knockout mice was higher than that in normal mice. The relative luciferase activities were enhanced by spermine. Conclusion Spermine increased the Atp8al gene expression at the mRNA level in Azinl knockout mice by enhancing promoter activity.