ROLE OF RECEPTORS IN ESTROGEN ACTION IN THE UTERUS

受体在子宫雌激素作用中的作用

• 批准号: 3313135
• 负责人: EUGENE R DE-SOMBRE
• 金额: $ 20.56万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-08-01 至 1990-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3313135
• 关键词: autoradiography; chromatin; gel electrophoresis; gel filtration chromatography; gene expression; genetic regulation; glucocorticoids; growth /development; hormone regulation /control mechanism; molecular pathology; progestins; radiotracer; scintillation spectrometry; tritium; uterus

The objective is to understand the role of estrogen receptors in the regulation of uterine growth and function by elucidating the important receptor changes which occur concomitantly or are required for estrogen regulation of gene expression in target cells. This is to be accomplished by studying the steroid hormone-regulated expression of several recently characterized estrogen-dependent secretory rat endometrial proteins, correlated with estrogen-dependent receptor modulation in the same cells, documenting changes in the receptor protein with a library of monoclonal antibodies to the estrogen receptor as well as steroid binding methods. The estrogen-responsive, marker proteins of the rat endometrial epithelium will be isolated, purified and characterized and used for preparation of polyclonal and monoclonal antibodies. These marker protein antibodies will be used to identify the location and possible function of the proteins and to isolate the specific m-RNAs for the proteins. After preparation of c-DNA, detailed studies will characterize the nature of the regulation of the marker at the level of transcription and translation. The c-DNA will eventually be used to screen genomic libraries to identify the gene for the estrogen-dependent markers so that the molecular details of the receptor regulated expression of the marker can provide new insight into the estrogen regulation of gene expression in the uterus. Immunoblots, binding and fractionation studies as well as immunohistochemical assays with the monoclonal antibodies to the receptor will be used to characterize changes in the nature, content and localization of the estrogen receptor in the different types of uterine cells under various hormonal conditions. Immunomethods will be used to study the major receptor events, such as estrogen receptor synthesis, potentiation, activation, processing, recycling, degradation and receptor binding to chromatin and nuclear matrix. These studies can be expected to provide new information on changes in the estrogen receptor, not available simply from assay of estrogen-binding studies, and to improve our understanding of the molecular details of steroid hormone regulation of the individual cell types of the uterus at the molecular level. This new knowledge will be useful for better diagnosis and treatment of steroid hormone-dependent cancers and will provide useful information for the control of normal and abnormal reproductive function.

目的是了解雌激素受体在卵巢癌中的作用。 通过阐明子宫发育和功能的重要性来调节子宫的生长和功能 伴随发生的或雌激素所必需的受体变化 靶细胞中基因表达的调控。这是要完成的 通过研究最近几个类固醇激素调控的基因表达 以雌激素依赖的分泌型大鼠子宫内膜蛋白为特征， 与同一细胞中雌激素依赖受体的调节相关， 用单克隆库记录受体蛋白的变化 雌激素受体抗体以及类固醇结合方法。 大鼠子宫内膜上皮雌激素反应标志蛋白的研究 将被分离、纯化和表征，并用于制备 多克隆抗体和单抗。这些标志性蛋白抗体将 用来确定蛋白质的位置和可能的功能 分离蛋白质的特异性m-RNAs。在准备好后 C-DNA，详细的研究将表征调控的性质 转录和翻译水平上的标记。C-DNA将 最终用于筛选基因组文库，以鉴定该基因 雌激素依赖的标志物使受体的分子细节 该标记物的调节表达可以提供对 雌激素对子宫基因表达的调节。免疫印迹，结合 和分级研究以及免疫组织化学分析 针对受体的单抗将被用来表征变化。 在雌激素受体的性质、含量和定位中 不同激素条件下不同类型的子宫细胞。 免疫方法将用于研究主要的受体事件，如 雌激素受体的合成，增强，激活，加工， 再循环、降解和受体与染色质和核的结合 矩阵。这些研究可望提供有关 雌激素受体的变化，不能简单地从 雌激素结合的研究，并提高我们对分子的理解 类固醇激素对不同细胞类型的调节细节 在分子水平上的子宫。这一新知识将对 更好地诊断和治疗类固醇激素依赖型癌症和 将为正常和异常的控制提供有用的信息 生殖功能。