摘要
目的:鉴定雌激素上调LRP16基因表达关键的靶启动子区域并探讨其分子机制。方法:对LRP16基因5′-侧翼2.6kb中的-213bp至-24bp启动子区进行亚克隆,构建5个控制不同启动子长度的荧光素酶报告重组子pS7-11,以既往构建的pS5、pS6为对照,与雌激素受体α(ERα)真核表达载体共转染MCF-7和Hela细胞,加入雌二醇(17β-E2)刺激后通过Luciferase assay方法测定相对荧光素酶活性。结果:MCF-7和Hela两个细胞系各组相对荧光素酶活性值上升趋势相平行,-213bp至-24bp区域各启动子克隆均具有较高的转激活活性,特别是pS10,在MCF-7细胞中对报告基因的上调活性达427倍,远远高于pS5。结论:E2主要通过-213bp至-24bp区域增强LRP16基因的转录,其中-213bp至-126bp(pS10)应为关键的启动子区。
Objective：To identify the primary promoter of LRP16 gene and explore the possible regulation mechanism of LRP16 gene expression up-regulated by estrogen. Methods：The human LRP16 gene promoter, -213 to -24bp of 5′-flanking DNA fragment （2.6kb）, were sub-cloned and inserted into pGL3-Basic vectors. Then the five constructs pS7-11 plus two previous constructs pS5 and pS6 were transient cotransfection with estrogen receptoret （ERot） gene expression vector in MCF-7 and Hela cells. The relative Luciferase activity induced by 17β-estradiol （β-E2） was detected by Luciferase Assay. Results： All constructs can up-regulate the relative luciferase activity in MCF-7 and Hela cells cotransfected with ERα expression vector, and the increased extent in MCF-7 and Hela cells was nearly consistent. The E2-induced transactivation activity conferred by pS10 reach 427 times, was significantly increased than those conferred by pS5. Conclusion：LRP16 gene expression is induced by E2 via the region of -213 to -24 bp, and the fragment of -213bp to -126bp（pS10）may be the primary promoter.
出处
《军医进修学院学报》
CAS
北大核心
2006年第3期161-163,共3页
Academic Journal of Pla Postgraduate Medical School
基金
国家自然科学基金项目(30471990)
北京市自然科学基金项目(5052024)
