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: 7583966
• 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/7583966
• 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; Prevention; Protein Isoforms; 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; mammary gland development; 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.

该研究计划的长期目标是阐明其功能和潜在机制 雌激素信号在乳腺和乳腺正常发育生物学中的作用 肿瘤发生。 在我们的初步研究中，我们鉴定并克隆了雌激素受体 (ER) 的 36 kDa 新变体 -a (ER-a36) 介导膜启动的雌激素和抗雌激素信号传导。它抑制基因组 ER-a 和-(3 介导的雌激素信号传导。ER-a36 在 60% 的乳腺癌标本中表达 进行了检查，甚至在 79% 的肿瘤中也发现了 ER 阴性乳腺癌。我们还发现 乳腺癌易感基因 1 (BRCA1) 负向调节 ER-a36 表达。我们将追求这些 通过以下实验方法进一步进行初步研究。 1. 我们将研究 ER-cc36 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 阴性乳腺癌细胞和乳腺转化中观察到 使用体外和体内细胞生长和转化测定结合 BRCA1 功能障碍 siRNA 敲低方法。 2. 我们将研究ER-cc36在抗雌激素的恶性生长和发育中的功能 使用表达不同水平 ER-a36 的 MCF7 细胞变体来治疗 ER 阳性乳腺癌的耐药性。 我们还将研究 ER-a36 在雌激素/抗雌激素信号传导中发挥作用的机制 微阵列方法和生物化学测定。 3. 我们将研究ER-a36调节ER-a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-a36 的表达受 ER-a、MARK 通路和 BRCA1 通过结构和 使用 DNA-蛋白质结合测定和荧光素酶测定对 ER-a36 启动子进行功能分析。 这些研究的意义在于它们将提供重要且新颖的信息 关于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。 这些知识将极大地推动我们在预防和治疗人类乳腺癌方面取得进展。