CAJ | 학술논문

目的：我们前期研究中进行全基因组表达谱检测发现在雄激素受体(androgen receptor, Ar)基因睾丸支持细胞特异性敲除小鼠( S-Ar-/ y )睾丸组织中泛素特异性蛋白酶25(ubiquitin specific peptidase 25, Usp25)基因表达较低。本研究的目的是了解雄激素及其受体是否可以作用于 Usp25基因,并测定其雄激素反应元件(androgen-responsive element, ARE)。方法采用 RT-qPCR 方法检测 Usp25基因表达量。通过生物信息学预测 Usp25基因上游可能的 ARE,构建 Usp25基因 ARE 报告质粒 pGLP/ Usp25。在 TM4细胞中,采用荧光素酶报告系统分析雄激素及其受体对 Usp25基因 ARE 活性的调控作用。结果在 S-Ar-/ y小鼠睾丸组织中 Usp25基因表达量比在野生型小鼠中显著降低。在 TM4细胞中睾酮可以显著提高 Usp25基因表达量。在 TM4细胞中,雄激素可以显著提高 pGLP/ Usp25的荧光素酶活性。结论 Usp25基因表达可以被雄激素睾酮激活, Usp25基因第一内含子519至1102 bp 区域含有 ARE,可以调控启动子的转录水平。
목적：아문전기연구중진행전기인조표체보검측발현재웅격소수체(androgen receptor, Ar)기인고환지지세포특이성고제소서( S-Ar-/ y )고환조직중범소특이성단백매25(ubiquitin specific peptidase 25, Usp25)기인표체교저。본연구적목적시료해웅격소급기수체시부가이작용우 Usp25기인,병측정기웅격소반응원건(androgen-responsive element, ARE)。방법채용 RT-qPCR 방법검측 Usp25기인표체량。통과생물신식학예측 Usp25기인상유가능적 ARE,구건 Usp25기인 ARE 보고질립 pGLP/ Usp25。재 TM4세포중,채용형광소매보고계통분석웅격소급기수체대 Usp25기인 ARE 활성적조공작용。결과재 S-Ar-/ y소서고환조직중 Usp25기인표체량비재야생형소서중현저강저。재 TM4세포중고동가이현저제고 Usp25기인표체량。재 TM4세포중,웅격소가이현저제고 pGLP/ Usp25적형광소매활성。결론 Usp25기인표체가이피웅격소고동격활, Usp25기인제일내함자519지1102 bp 구역함유 ARE,가이조공계동자적전록수평。
Objective It was found that the expression of ubiquitin specific peptidase 25 (Usp25) gene was decreased in the testes of Sertoli cellArknockout (S-Ar-/ y ) mice. The upregula-tion ofUsp25gene by testosterone and AR was further demonstrated and androgen-responsive element ( ARE ) of theUsp25was validated. Methods The expression ofUsp25was detected by RT-qPCR. Bioinformatics methods were used to predict the potential ARE ofUsp25gene. The mouseUsp25ARE recombinant of pGLP was constructed as a reporter plasmid. Then, the activation effects of testosterone and androgen receptor on theUsp25ARE were analyzed by the fluorescent re-porter system in TM4 cells. Results The expression ofUsp25gene was significantly decreased in the testes of S-Ar-/ y mice and upregulated by testosterone in TM4 cells. The luciferase activity of pGLP/Usp25 was also upregulated by testosterone in TM4 cells. Conclusion TheUsp25gene expression can be activited by testosterone. There is ARE in the sequence from 519 to 1102 of the first intron ofUsp25gene.