HOXA10与连续TTAT基序结合抑制p/CAF表达  

HOXA10 suppresses p/CAF expression via binding to three consecutive TTAT units

作　　者：华美娟[1,2] 赵静[2] 陈林君[2] 颜桂军[2] 刁振宇[2] 胡娅莉[1,2] 孙海翔[1,2] 

机构地区：[1]南京医科大学鼓楼临床医学院,江苏南京210029 [2]南京大学医学院附属鼓楼医院生殖医学中心,江苏南京210008

出　　处：《医学研究生学报》2009年第11期1143-1146,1149,I0003,共6页Journal of Medical Postgraduates

基　　金：江苏省"兴卫工程"医学重点学科资助基金(批准号:XK200709)

摘　　要：目的:转录因子HOXA10在胚胎着床和子宫内膜蜕膜化过程中发挥重要作用。p/CAF是HOXA10新的靶基因,文章旨在筛选并鉴定p/CAF启动子区与HOXA10功能性结合的序列。方法:制备重组腺病毒HOXA10(ad-HOXA10),用于感染人子宫内膜间质细胞(hESC);构建p/CAF启动子区多种突变报告基因质粒;采用萤光素酶报告基因实验检测结合腺病毒介导的HOXA10基因过表达,分析hESC中HOXA10对p/CAF启动子转录活性的调控。结果:获得滴度为2.4×1011ifu/mL的重组ad-HOXA10,对hESC的感染率达到80%以上。HOXA10通过结合连续的3个TTAT(-710^-674 nt)基序抑制p/CAF启动子活性,连续的3个TTAT基序的一级结构影响HOXA10的功能性结合。结论:p/CAF启动子区连续的3个TTAT基序是HOXA10功能性结合所必须的,HOXA10可能通过调控p/CAF的表达来控制hESC的增殖与分化。Objective： HOXA10, a homeobox transcription factor, plays a key role in-embryo implantation and endometrial decidualization. The p300/CBP-associated factor （p/CAF） is a novel HOXA10 target gene identified by CHIP assay. This study aimed to identify the functional binding sites of HOXA10 in the p/CAF promoter region. Methods ： Adenovirus-HOXA10 was prepared to infect human endometrial stromal cells （hESCs） and generate a series of promoter-Luc reporter plasmids in the p/CAF promoter region. The role of HOXA10 in the regulation of p/CAF promoter activity in the hESCs was analyzed by luciferase reporter assay and adenovirus-mediated overexpression of HOXA10. Results： We obtained 2.4 × 10^11 ifu/mL ad-HOXA10 and an 80% infection of the hESCs. HOXA10 suppressed the p/CAF promoter activity via binding to three consecutive TYAT elements （ -710 - -674 nt）, whose primary structures influenced the functional binding of HOXA10. Conclusion： Three consecutive TTAT elements within the p/CAF promoter are required for I-IOXA10 functional binding. HOXAI0 may regulate the proliferation and differentiation of hESCs bv changing the expression of B/CAF.

关 键 词：HOXA10 p/CAF 人子宫内膜间质细胞 

分 类 号：Q756[生物学—分子生物学]