Estrogen Signaling in Normal and Transformed Cell Growth

正常和转化细胞生长中的雌激素信号传导

• 批准号: 7104684
• 负责人: ZHAO-YI WANG
• 金额: $ 29.42万
• 依托单位: CREIGHTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-03-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7104684
• 关键词: MCF7 cell; biological signal transduction; brca gene; breast neoplasms; chromatin immunoprecipitation; estrogen receptors; estrogens; gel mobility shift assay; gene targeting; mammary gland; microarray technology; mitogen activated protein kinase; neoplastic cell; neoplastic growth; neoplastic transformation; protein structure function; receptor expression; reproductive development; small interfering RNA

DESCRIPTION (provided by applicant): The long-term goals of this research program are to elucidate the functions and underlying mechanisms of estrogen signaling in the biology of normal development of mammary glands and mammary tumorigenesis. In our preliminary studies, we identified and cloned a 36-kDa novel variant of Estrogen Receptor (ER)-a (ER-a36) that mediates membrane-initiated estrogen-and antiestrogen-signaling. It inhibits genomic estrogen signaling mediated by ER-a and-(3. ER-a36 is expressed in 60% breast cancer specimens examined, and even in 79% tumors characterized as ER-negative breast cancers. We also found that breast cancer susceptibility gene 1 (BRCA1) negatively regulates ER-a36 expression. We will pursue these preliminary studies further through the following experimental approaches. 1. We will study the functions of ER-cc36 in membrane-initiated estrogen- and antiestrogen-signaling observed in established ER-negative breast cancer cells and in mammary transformation resulted from BRCA1 dysfunction using in vitro and in vivo cell growth and transformation assays combined with the siRNA knockdown approach. 2. We will study the functions of ER-cc36 in malignant growth and development of antiestrogen resistance in ER-positive breast cancer using MCF7 cell variants that express different levels of ER-a36. We will also study the mechanisms by which ER-a36 functions in estrogen/antiestrogen signaling using the microarray approach and biochemistry assays. 3. We will study the mechanisms by which ER-a36 regulates ER-a expression and function using biochemistry, the gel shift, ChIP and luciferase assays. We will also study the molecular mechanisms by which ER-a36 expression is regulated by ER-a, the MARK pathway and BRCA1 through structure and function analysis of the ER-a36 promoter using DNA-protein binding assays and the luciferase assay. The significance of these studies resides in the fact that they will provide important and novel information on the molecular mechanisms by which estrogen signaling functions in normal and transformed cell growth. Such knowledge will greatly advance our progress in prevention and treatment of human breast cancer.

描述（由申请人提供）：该研究计划的长期目标是阐明雌激素信号传导在乳腺正常发育和乳腺肿瘤发生的生物学中的功能和潜在机制。在我们的初步研究中，我们确定并克隆了雌激素受体（ER）-A（ER-A36）的36 kDa新颖变体，该变体介导了膜引起的雌激素和抗雌激素信号。它抑制了由ER-A介导的基因组雌激素信号传导和 - （3。ER-A36在检查的60％的乳腺癌标本中表达，甚至在79％的肿瘤中，以ER阴性乳腺癌为特征。我们还发现乳腺癌易感性基因1（BRCA1）负责ER-A36的陈述。 ER-CC36在膜引起的雌激素和抗雌激素信号中的功能在已建立的ER阴性乳腺癌细胞中观察到，在BRCA1功能障碍中，使用体外细胞的生长和体内转化分析与sirna敲低接近的功能相结合。使用MCF7细胞变体的乳腺癌表达不同水平的ER-A36。 3。我们将研究ER-A36使用生物化学，凝胶移位，芯片和荧光素酶测定法调节ER-A的表达和功能的机制。我们还将通过使用DNA-蛋白结合测定法和荧光素酶测定法调节ER-A，MARK途径和BRCA1的ER-A36表达，MARK途径和BRCA1的分子机制。这些研究的重要性在于它们将提供有关雌激素信号在正常和转化细胞生长中功能的分子机制的重要和新颖的信息。这种知识将大大推动我们在预防和治疗人类乳腺癌方面的进步。