雌激素上调LRP16基因表达靶启动子区的研究  

Study on the target promoter of LRP16 gene expression upregulated by estrogen

作　　者：司艺玲[1] 韩为东[1] 赵亚力[1] 李琦[1] 李雪[1] 宋海静[1] 母义明[2] 

机构地区：[1]解放军总医院基础医学所分子生物室,北京100853 [2]解放军总医院内分泌科,北京100853

出　　处：《军医进修学院学报》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)

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.

关 键 词：雌激素 基因表达调控 LRP16 启动区(遗传学) 

分 类 号：R349.64[医药卫生—基础医学]