MANIPULATING GLUCOCORTICOID RECEPTOR IN LIVE TUMOR CELLS

我的长期目标是了解 受体介导的糖皮质激素作用。糖皮质激素是 既有免疫抑制作用，又有抗炎作用 作为治疗白血病的药物和治疗工具 淋巴瘤。目前，关于以下方面的知识还远远不完整 激素敏感型肿瘤细胞的病因和/或控制 类固醇在肿瘤中诱导反应的分子机制 在正常细胞上，或一些细胞逃避荷尔蒙的途径 控制力。这在一定程度上是由于传统方法的性质。 类固醇受体的研究，因为他们依赖于间接的 需要形成和维护的检测战略 类固醇受体复合体，通常在细胞- 自由系统。该项目的目的是开发和开发 评估一个新的实验系统，在这个系统中，分子和 将直接探索类固醇激素作用的细胞生物学 在活细胞中。即将引入的生物探针 系统是1)结合到一系列 糖皮质激素受体的表位和修饰功能， 2)免疫亲和力纯化的完整受体及其 包含特定功能结构域的离散片段。一个 基于红细胞幽灵介导法的精化策略 会被用来在生物上“注射”大量这样的物质 活性大分子进入糖皮质激素的批量培养中- 反应灵敏和耐药的肿瘤细胞。按照规定，这一点 实验系统具有显著的通用性，因此将 使体内受体中和、竞争、 以及任何可用的糖皮质激素的替代研究- 响应型和变异型抗性细胞系。抗体库I 希望发展也应该提供强大的新的基础 分析程序，并允许对结构和 糖皮质激素受体的功能，特别是类固醇 一般情况下，受体。

My long-term objective is to understand the mechanism of receptor-mediated glucocorticoid action. Glucocorticoids are important both as immunosuppressive and anti-inflammatory agents and as therapeutic tools in the treatment of leukemias and lymphomas. Currently, knowledge is far from complete regarding the etiology and/or control of steroid-sensitive tumor cells, the molecular mechanism of steroid-induced responses in neoplastic on normal cells, or the route by which some cells escape hormonal control. This is due in part to the nature of traditional methods of steriod receptor study in that they have relied on indirect detection strategies requiring the formation and maintenance of steroid-receptor complexes, and are usually performed in cell- free systems. The purpose of this project is to develop and evaluate a novel experimental system in which the molecular and cellular biology of steriod hormone action will be directly probed in living cells. The biological probes to be introduced into the system are 1) monoclonal antibodies that bind to an array of epitopes on, and modify function of the glucocorticoid receptor, and 2) the immunoaffinity-purified intact receptor and its discrete fragments containing specific functional domains. A refined strategy based on the erythrocyte ghost-mediated method will then be used to "inject" large numbers of these biologically active macromolecules into bulk cultures of glucocorticoid- responsive and resistant tumor cells. As formulated, this experimental system possesses marked versatility, and thus will make feasible novel in vivo receptor neutralization, competition, and replacement studies in any of the available glucocorticoid- responsive and variant resistant cell lines. The antibody bank I hope to develop should also provide the basis of powerful new analytical procedures and allow new insight into the structure and function of the glucocorticoid receptor in particular, and steroid receptors in general.