Transcriptional activation of steroidogenic factor-1 by hypomethylation of the 5′ CpG island in endometriosis
Transcriptional activation of steroidogenic factor-1 by hypomethylation of the 5′ CpG island in endometriosis
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DOI：
10.1210/jc.2007-0494
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发表时间：
2007-08-01
影响因子：
5.8
通讯作者：
Bulun, Serdar E.
中科院分区：
文献类型：
--
作者：
Xue, Qing;Lin, Zhihong;Bulun, Serdar E.
背景：子宫内膜异位症是一种雌激素依赖性疾病。甾体生成因子-1 （SF-1）是激活雌激素生物合成的多种甾体生成基因所必需的转录因子，在正常子宫内膜间质细胞中检测不到，而在子宫内膜异位症间质细胞中异常表达。目的：研究SF-1在子宫内膜和子宫内膜异位症间质细胞中的表达差异机制。设计：我们确定了一个位于SF-1启动子和外显子I区域两侧的CpG岛，并确定了其在子宫内膜和子宫内膜异位症细胞中的甲基化模式。环境：该研究在美国西北大学进行。患者或其他参与者：研究来自无疾病受试者的异位子宫内膜（n = 8）和卵巢囊性子宫内膜异位症病变的壁（n = 8）。干预：从这两种组织中分离基质细胞。主要结果测量指标：测量指标在结果中提及。结果：子宫内膜异位症间质细胞中SF-1 mRNA和蛋白水平显著高于子宫内膜间质细胞（P < 0.001）。亚硫酸氢盐测序显示，与子宫内膜异位症细胞相比，子宫内膜细胞的甲基化显著增加（P < 0.001）。5-aza-2(')-脱氧胞苷去甲基化可使子宫内膜细胞中SF-1 mRNA水平升高55.48倍（P < 0.05）。荧光素酶检测结果显示，CpG岛所在的-85/+239区调控其活性（P < 0.01）。该区域的自然或体外甲基化显著降低了两种细胞类型中SF-1启动子的活性（P < 0.01）。染色质免疫沉淀试验表明，甲基cpg结合域蛋白2在子宫内膜中与SF-1启动子结合，而在子宫内膜异位症细胞中不与SF-1启动子结合。结论：这是首次在任何哺乳动物组织中证明了SF-1的甲基化依赖性调节。这些发现指出了子宫内膜异位症靶向局部雌激素生物合成的新机制。
Context: Endometriosis is an estrogen-dependent disease. Steroidogenic factor-1 (SF-1), a transcriptional factor essential for activation of multiple steroidogenic genes for estrogen biosynthesis, is undetectable in normal endometrial stromal cells and aberrantly expressed in endometriotic stromal cells.Objective: The objective of the study was to unravel the mechanism for differential SF-1 expression in endometrial and endometriotic stromal cells.Design: We identified a CpG island flanking the SF-1 promoter and exon I region and determined its methylation patterns in endometrial and endometriotic cells.Setting: The study was conducted at Northwestern University.Patients or Other Participants: Eutopic endometrium from diseasefree subjects (n = 8) and the walls of cystic endometriosis lesions of the ovaries (n = 8) were investigated. Intervention(s): Stromal cells were isolated from these two types of tissues.Main Outcome Measure(s): Measures are mentioned in Results.Results: SF-1 mRNA and protein levels in endometriotic stromal cells were significantly higher than those in endometrial stromal cells (P < 0.001). Bisulfite sequencing showed strikingly increased methylation in endometrial cells, compared with endometriotic cells (P < 0.001). Demethylation by 5-aza-2(')-deoxycytidine increased SF-1 mRNA levels by up to 55.48-fold in endometrial cell (P < 0.05). Luciferase assays showed that the -85/+239 region bearing the CpG island regulated its activity (P < 0.01). Natural or in vitro methylation of this region strikingly reduced SF-1 promoter activity in both cell types (P < 0.01). Chromatin immunoprecipitation assay showed that methyl-CpG-binding domain protein 2 binds to the SF-1 promoter in endometrial but not endometriotic cells.Conclusions: This is the first demonstration of methylation-dependent regulation of SF-1 in any mammalian tissue. These findings point to a new mechanism for targeting local estrogen biosynthesis in endometriosis.