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作 者:赵静[1,2] 华美娟[2] 陈林君[2] 颜桂军[2] 李朝军[1] 孙海翔[2]
机构地区:[1]南京师范大学生命科学学院,江苏省分子医学生物技术重点实验室,江苏南京210046 [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:《生物技术通讯》2009年第5期612-615,共4页Letters in Biotechnology
摘 要:目的:筛选与鉴定转录因子同源框蛋白(HOX)A10下游靶基因。方法:用Ad-HOXA10重组腺病毒感染人子宫内膜间质细胞,通过染色体免疫共沉淀(ChIP)方法,筛选HOXA10的下游靶基因;采用萤光素酶报告基因实验结合腺病毒介导的HOXA10过表达和小干扰RNA介导的基因沉默实验,分析HOXA10对下游靶基因的转录调控作用。结果:ChIP-PCR筛选并鉴定p/CAF(p300/CBP相关因子)为HOXA10直接作用的靶基因;过表达HOXA10抑制p/CAF启动子活性达60%;基因沉默内源性HOXA10的表达可以激活p/CAF启动子活性超过2倍以上。结论:p/CAF是HOXA10新的靶基因,HOXA10可能通过调节p/CAF的表达来调控子宫内膜的发育。Objective: To screen and identify the target genes of transcription factor homeobox protein(HOX) A10. Methods: Using a chromatin immunoprecipitation screen to identify the target genes of HOXA10 in human endometrial stromal eells(hESCs). Luciferase reporter assay and adenovirus-mediated over-expression and siRNA-specific knockdown of HOX- AIO were performed to study the regulation of the target genes promoter activity by HOXA10 in hESCs. Results: The p300/CBP-associated factor(p/CAF) is a direct target of HOXA10 in hESCs. Over-expression of HOXA10 decreased luciferase activities by about 60% in hESCs. Knockdown of HOXAIO increased the basal p/CAF promoter activity by greater than twofold in hESCs. Conclusion: p/CAF is a novel HOXA10 target gene, and HOXA10 promotes human endometrial development, at least in part, through the regulation of p/CAF gene.
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