Estrogen receptor Alpha/Beta antagonism in osteoblasts

成骨细胞中雌激素受体α/β拮抗作用

• 批准号: 6444993
• 负责人: David G Monroe
• 金额: $ 4.37万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-18 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6444993
• 关键词: bone metabolism; cell line; estrogen receptors; estrogens; gene targeting; genetic transcription; genetically modified animals; hormone regulation /control mechanism; immunoprecipitation; laboratory mouse; microarray technology; osteoblasts; osteocytes; protein structure function; raloxifene; receptor expression; tamoxifen; transfection /expression vector; transforming growth factors; western blottings

DESCRIPTION (provided by applicant): Estrogen is a key regulator in the growth and maintenance of bone mass in both sexes. Two isoforms of estrogen receptor, ERalpha and ERbeta, mediate the transcriptional effects of estrogen by binding 1 7-beta.estradiol (E2) and subsequently binding to specific elements within the regulatory regions of genes. Although we, and others, have shown that ERalpha and ERbeta are functional in osteoblasts, little data are available concerning the interactions of ERalpha and ERbeta in bone. Recent gene deletion experiments in mice suggest that the ERalpha and ERbeta isoforms may have opposing actions on bone, by analogy with the A and B isoforms of PR, where PR-A is an inhibitor of PR-B. One mechanism to explain the potential antagonistic effects of ERalpha and ERbeta would involve the differential recruitment of steroid receptor coactivators (SRCs), which transmit the transcriptional signal to the basal transcriptional machinery. Therefore, the purpose of this research proposal is to understand the functions of ERalpha/alpha and ERbeta/beta homodimers in human osteoblasts and to understand the effects of the ERalpha/beta heterodimer on gene transcription in human osteoblasts. The project will attempt to address these basic biological questions using three approaches: 1) to understand the effects of ERalpha/beta heterodimers on the transcriptional potential at canonical estrogen response elements (EREs) and at SP1/(ERE1/2) sites in hFOB and MG-63 cell lines; 2) determine the recruitment of steroid receptor coactivators (SRCs) by various ER homo- and heterodimers; and finally 3) determine the in vivo consequences of ERalpha/alpha, ERbeta/beta and ERalpha/beta expression in human OB cells using gene chip technology.

描述(申请人提供)：雌激素是生长的关键调节剂。 并维持两性的骨量。雌激素受体的两种异构体， ERα和ERβ通过结合介导雌激素的转录效应 1，7-β-雌二醇(E2)，并随后与 基因的调控区域。尽管我们和其他人已经证明了 ERa和ERbeta在成骨细胞中具有功能，但数据很少 关于ERa和ERbeta在骨骼中的相互作用。最近的基因缺失 在小鼠身上的实验表明，ERpha和ERbeta亚型可能具有 对骨骼的相反作用，类推PR的A和B亚型，其中 PR-A是PR-B的抑制剂。一种解释这种潜力的机制 ERpha和ERbeta的拮抗作用将涉及到差异 类固醇受体共激活剂(SRCs)的招募，它传递 转录信号传递给基本的转录机制。因此， 本研究建议的目的是为了了解 人成骨细胞中ERα/α和ERbeta/β同源二聚体的表达及意义 ERα/β异源二聚体对人类基因转录的影响 成骨细胞。该项目将试图解决这些基本生物学问题 使用三种方法提问：1)了解ERAlpha/Beta的影响 异源二聚体对典型雌激素反应转录潜能的影响 HFOB和MG-63细胞中的元件(ERE)和SP1/(ERE1/2)位点；2) 测定不同内质网对类固醇受体共激活因子(SRCs)的募集 同源和异源二聚体；最后3)决定了在体内的后果 人卵巢上皮细胞ERα/α、ERbeta/β和ERAlpha/β的表达 基因芯片技术。