Coactivators and Androgen Receptors in Prostate Cancer

前列腺癌中的共激活剂和雄激素受体

• 批准号: 7057211
• 负责人: Nancy L. Weigel
• 金额: $ 33.07万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7057211
• 关键词: androgen receptor; biological signal transduction; carcinogenesis; cell growth regulation; gene expression; genetic promoter element; genetic regulation; genetic transcription; hormone regulation /control mechanism; immunoprecipitation; laboratory mouse; ligands; male; posttranslational modifications; prostate neoplasms; receptor binding; receptor expression

DESCRIPTION (provided by applicant): Prostate cancer is initially an androgen dependent disease and there is mounting evidence that the androgen receptor (AR) continues to play a role in androgen ablation resistant prostate cancer. Great strides have been made in recent years in elucidating the structure of AR, identifying candidate coregulatory proteins, and in identifying some of the genes regulated by AR. Nonetheless, our understanding of the mechanism of action of AR and our knowledge of the identity of the key genes regulated by AR to induce growth of prostate cancer cells is insufficient to provide a good basis for rational development of alternate means of blocking AR activity. Our understanding of and ability to effectively block the growth of androgen independent prostate cancer is even less well advanced. Although many candidate AR coactivators have been identified by molecular biological techniques, virtually nothing is known about their individual roles in cells, their contributions to positive and negative regulation of AR target genes and to AR dependent cell growth. Moreover, their role in androgen independent AR action has not been determined. To address these issues, we propose: Specific Aim 1: To test the hypothesis that the androgen receptor plays a key role in the growth and gene expression in some androgen independent prostate cancer cells as well as in androgen dependent prostate cancer cells and to evaluate the role of receptor coactivators in both hormone dependent and independent cell growth and gene regulation. We will selectively reduce expression of candidate coactivators and assess the effects on cell growth and target gene expression. Specific Aim 2: To identify genes regulated by androgen receptor and to test the hypothesis that coactivator requirements are target gene/promoter specific. We will utilize two approaches, Affymetrix U133A chips and a novel Protein A-AR chimera containing a TEV (tobacco etch virus) protease site linker to identify AR target genes and to evaluate the consequences of coactivator reduction on target gene expression. Specific Aim 3: To utilize chromatin immunoprecipitation (CHIP) assays to identify coregulators involved in hormone dependent and ligand independent transcriptional regulation by androgen receptors and to assess the consequences of elimination of specific coregulators on the composition of proteins associated with promoters and on the post-translational modifications of proteins associated with the promoter.

描述（由申请人提供）： 前列腺癌最初是一种雄激素依赖性疾病，并且越来越多的证据表明雄激素受体（AR）继续在雄激素消融抵抗性前列腺癌中发挥作用。近年来，在阐明 AR 结构、鉴定候选共调节蛋白以及鉴定一些受 AR 调节的基因方面取得了长足的进步。尽管如此，我们对AR作用机制的理解以及对AR调节诱导前列腺癌细胞生长的关键基因的了解不足以为合理开发阻断AR活性的替代方法提供良好的基础。我们对有效阻止雄激素非依赖性前列腺癌生长的理解和能力甚至还不够先进。尽管许多候选的 AR 共激活剂已通过分子生物学技术鉴定出来，但实际上对它们在细胞中的各自作用、它们对 AR 靶基因的正向和负向调节以及对 AR 依赖性细胞生长的贡献一无所知。此外，它们在不依赖于雄激素的 AR 作用中的作用尚未确定。为了解决这些问题，我们提出： 具体目标 1：检验雄激素受体在一些雄激素非依赖性前列腺癌细胞以及雄激素依赖性前列腺癌细胞的生长和基因表达中发挥关键作用的假设，并评估受体共激活剂在激素依赖性和非依赖性细胞生长和基因调节中的作用。我们将选择性地减少候选共激活剂的表达，并评估对细胞生长和靶基因表达的影响。具体目标 2：鉴定受雄激素受体调节的基因，并检验共激活因子需求是靶基因/启动子特异性的假设。我们将利用两种方法，Affymetrix U133A 芯片和含有 TEV（烟草蚀刻病毒）蛋白酶位点连接子的新型蛋白 A-AR 嵌合体来识别 AR 靶基因并评估共激活剂减少对靶基因表达的影响。具体目标 3：利用染色质免疫沉淀 (CHIP) 测定来鉴定参与雄激素受体的激素依赖性和配体非依赖性转录调节的辅助调节因子，并评估消除特定辅助调节因子对与启动子相关的蛋白质组成和后转录调节的影响。与启动子相关的蛋白质的翻译修饰。