ESTROGEN STRUCTURE-RECEPTOR FUNCTION RELATIONSHIPS

雌激素结构-受体功能关系

• 批准号: 3187557
• 负责人: SAMUEL C BROOKS
• 金额: $ 28.72万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-04-15 至 1994-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3187557
• 关键词: DNA binding protein; chemical structure function; density gradient ultracentrifugation; estrogen analog; estrogen receptors; gene expression; genetic regulation; genetic transcription; hormone regulation /control mechanism; messenger RNA; nuclear runoff assay; plasminogen activator; progesterone receptors; protein structure; protein structure function; receptor binding; tissue /cell culture

DESCRIPTION (Adapted from the applicant's abstract): Initial studies have shown that the structure of the estrogen ligand can dictate the range of genes induced and the concentration limits for the ligand to stimulate an estrogen response in MCF-7 human breast cancer cells. Structural modifications to the 17beta-estradiol (E2) molecule in the A and D rings are influential in discriminating among the range of estrogen responsive genes. The proposed studies are designed to first evaluate the effects of these E2 derivatives on induction of four E2-responsive genes (pS2, progesterone receptor, tissue plasminogen activator, and cathepsin D) and MCF-7 cell growth. Further "mapping" of the estrogen molecule for other components which may be important in the induction of responsive genes will be carried out with a series of B-and C-ring based derivatives of E2. Studies of the aberrant ER complex formed with E2 analogues will be pursued by documenting structural alterations in the analogue-receptor complex in solution by sucrose density gradient analysis and when bound to the estrogen responsive element (ERE) by gel retardation analysis. It is hypothesized that ER complexed with structurally altered estrogens may bind with different affinities and/or conformations to the consensus ERE and functional EREs that vary from it by one or more nucleotides. Furthermore, the possibility that analogue-ER complexes may be interacting with other regulatory elements known to influence estrogen responsive gene expression will be considered. Investigations concerned with the gene-specific induction exhibited by E2 isomers will be pursued by examining the ability of each estrogen analogue to activate the CAT gene in MCF-7 cells transfected with plasmid constructs in which CAT expression is regulated by different EREs or other regulatory elements. The abnormal activity of specific estrogen analogues will also be examined in experiments which document variances in transcription rate, half-life of mRNAs of gene products, and the effect of estrogen structure on ER dimer formation. Knowledge gained from these studies will contribute to the understanding of gene regulation by estrogens and aid in the design of attenuated estrogens for use in the treatment of hormone dependent neoplasms.

描述（根据申请人的摘要改编）：初始研究 表明雌激素配体的结构可以决定 诱导的基因和配体的浓度极限刺激 MCF-7人类乳腺癌细胞中的雌激素反应。 结构 对A和D环中的17beta-雌二醇（E2）分子的修饰 对雌激素反应范围的区分有影响 基因。 拟议的研究旨在首先评估 这些E2衍生物诱导了四个E2响应基因（PS2， 孕酮受体，组织纤溶酶原激活剂和组织蛋白酶D）和 MCF-7细胞生长。 进一步的雌激素分子的“映射” 可能在诱导反应基因的组件将 用一系列基于B和C形的衍生物E2进行。 将研究由E2类似物形成的异常ER复合物的研究 通过记录模拟受体复合物中的结构变化 通过蔗糖密度梯度分析解决方案，并与 通过凝胶延迟分析，雌激素反应元件（ERE）。 这是 假设ER与结构变化的雌激素复合可能结合 对共识的不同亲和力和/或构造 与一个或多个核苷酸不同的功能性ER。 此外， 模拟-er复合物可能正在与其他相互作用的可能性 已知影响雌激素反应基因表达的调节元件 将被考虑。 与基因特异性有关的调查 E2异构体展示的归纳将通过检查能力来追求 每个雌激素类似物以激活MCF-7细胞中的CAT基因 用质粒构建体转染，其中CAT表达受到调节 不同的ERE或其他监管元素。 异常活性 在实验中也将检查特定的雌激素类似物 转录率的文档差异，基因mRNA的半衰期 产品，以及雌激素结构对ER二聚体形成的影响。 从这些研究中获得的知识将有助于理解 通过雌激素调节基因并有助于衰减的雌激素的设计 用于治疗依赖激素的肿瘤。