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: 7391742
• 负责人: ZHAO-YI WANG
• 金额: $ 27.99万
• 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/7391742
• 关键词: Agreement; Binding; Biochemistry; Biological; Biological Assay; Biology; Breast Cancer Cell; Cancer Etiology; Cancer Patient; Cancer-Predisposing Gene; Cell Proliferation; DNA-Binding Proteins; Data; Development; Dimerization; Epithelial Cell Proliferation; Epithelial Cells; Estrogen Antagonists; Estrogen Receptor Modulators; Estrogen Receptors; Estrogen receptor negative; Estrogen receptor positive; Estrogens; Functional disorder; Gel; Genes; Genomics; Goals; Growth; Growth and Development function; Human; In Vitro; Interferon Gamma Receptor Beta Chain; Introns; Knowledge; Ligand Binding Domain; Luciferases; MCF7 cell; Malignant - descriptor; Mammary Tumorigenesis; Mammary gland; Mediating; Membrane; Molecular; Molecular Mechanisms of Action; Neoplastic Cell Transformation; Pathway interactions; Personal Satisfaction; Prevention; Protein Isoforms; Rate; Regulation; Research; Research Personnel; Resistance; Risk; Role; Signal Transduction; Small Interfering RNA; Specimen; Structure; Therapeutic; Transcriptional Activation Domain; Variant; cell growth; cell transformation; design; in vivo; malignant breast neoplasm; novel; programs; promoter; research study; tumor; tumor progression

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.

本研究计划的长期目标是阐明功能和潜在机制 雌激素信号在乳腺和乳腺癌正常发育生物学中的作用 肿瘤发生 在我们的初步研究中，我们鉴定并克隆了一个36 kDa的雌激素受体（ER）-a的新变体， （ER-a36），其介导膜起始的雌激素和抗雌激素信号传导。它抑制基因组 由ER-α和-β介导的雌激素信号传导（3. ER-α 36在60%的乳腺癌标本中表达 检查，甚至在79%的肿瘤特征为ER阴性乳腺癌。我们还发现 乳腺癌易感基因1（BRCA 1）负调控ER-α 36表达。我们将继续努力， 通过以下实验方法进一步进行初步研究。 1.我们将研究ER-α 36在膜启动的雌激素和抗雌激素信号传导中的功能。 在已建立的ER阴性乳腺癌细胞中以及由ER阴性乳腺癌细胞引起的乳腺转化中观察到 使用体外和体内细胞生长和转化测定结合BRCA 1功能障碍， siRNA敲低方法。 2.本研究旨在探讨ER-cc36在恶性肿瘤发生、发展中的作用及抗雌激素作用。 使用表达不同水平的ER-a36的MCF7细胞变体在ER阳性乳腺癌中的耐药性。 我们还将研究ER-α 36在雌激素/抗雌激素信号传导中的作用机制， 微阵列方法和生物化学测定。 3.我们将研究ER-α 36调节ER-α表达和功能的机制， 生物化学、凝胶位移、ChIP和荧光素酶测定。我们还将研究分子机制， 其中ER-α 36表达通过结构由ER-α、MARK通路和BRCA 1调节， 使用DNA-蛋白质结合测定和荧光素酶测定进行ER-α 36启动子的功能分析。 这些研究的意义在于它们将提供重要而新颖的信息 雌激素信号在正常和转化细胞生长中发挥作用的分子机制。 这些知识将大大推动我们在预防和治疗人类乳腺癌方面的进展。