INTERACTIONS OF TUMOR PROMOTERS WITH RECEPTORS

肿瘤启动子与受体的相互作用

• 批准号: 3173795
• 负责人: CURTIS L. ASHENDEL
• 金额: $ 14.25万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-01-01 至 1989-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3173795
• 关键词: chemical carcinogen; chemical carcinogenesis; electron microscopy; membrane activity; phorbols; skin neoplasms; tumor promoters

The molecular mechanism of action of the tumor promoting phorbol esters will be investigated by determining the normal physiological function of the phorbol ester receptor protein in cells and the effect of tumor promoters on receptor function. Phorbol esters elicit numerous changes in cellular morphology, growth, and differentiation. This is generally assumed to occur via binding reversibly and with high affinity and specificity to a cellular receptor protein which has protein kinase activity tightly associated with it. Both membrane bound and soluble cytosolic receptors exist in cells although the latter requires calcium and phospholipid to function. This and other data suggest that as part of its normal function in cells the soluble cytosolic receptor becomes associated with cell membranes and that phorbol esters may directly affect this aspect of its function. The research plan involves analysis of changes in the localization of the receptor in stimulated cells by quantitation of phorbol ester binding activity, ultraviolet light microscopic observation of cells stained with fluorecently labeled receptor specific antibodies, and viewing of cells stained with receptor specific antibodies covalently bound to ferritin using a trasmission electron microscope. The antibodies will be produced by the monoclonal hybridoma method using as antigen receptor protein purified from mouse brain. Initial biochemical experiments will use as a model system primary mouse spleen cells stimulated with lectins, phorbol esters, and lymphokines. These results will be applied to a study of primary mouse epidermal keratinocytes and C3H 10T-1/2 fibroblasts in which growth and morphological controls are affected by calcium, polypeptide growth factors, and phorbol esters. Ultimately the investigation will be extended to mouse epidermis, in vivo, so that the effects of in vivo treatments known to affect the carcinogenic process in epidermis can be compared to the results obtained with cells in culture. These studies as well as a biochemical investigation of interactions of the receptor with other cellular proteins will lead to the development of a cell free system in which the actions of phorbol esters can be observed. These studies will help characterize the biological action of phorbol esters and aid in understanding the biological processes of tumor promotion and chemical carcinogenesis.

佛波醇酯促肿瘤作用的分子机制 将通过确定正常的生理功能进行研究， 细胞内佛波酯受体蛋白与肿瘤作用 启动子对受体功能的影响 佛波酯引起许多变化， 细胞形态、生长和分化。 这通常是 假设通过可逆结合发生，并具有高亲和力， 对具有蛋白激酶的细胞受体蛋白的特异性 活性紧密相关。膜结合和可溶性 胞质受体存在于细胞中，尽管后者需要钙， 磷脂发挥作用。 这一数据和其他数据表明，作为其 在细胞中的正常功能，可溶性胞质受体变得相关 与细胞膜和佛波醇酯可能直接影响这方面 它的功能。 研究计划包括分析 通过定量佛波醇在受刺激细胞中的受体定位 酯结合活性，细胞的紫外光显微镜观察 用荧光标记的受体特异性抗体染色，观察 用受体特异性抗体染色的细胞， 用透射电子显微镜观察铁蛋白。 抗体会 通过单克隆杂交瘤方法产生，用作抗原受体 从小鼠脑中纯化的蛋白质。 最初的生化实验将 使用凝集素刺激的原代小鼠脾细胞作为模型系统， 佛波醇酯和淋巴因子。 这些结果将应用于一项研究 原代小鼠表皮角质形成细胞和C3 H 10 T-1/2成纤维细胞在 其生长和形态控制受钙影响， 多肽生长因子和佛波醇酯。 最终 研究将扩展到小鼠表皮，在体内，以便 已知影响致癌过程的体内治疗的影响， 可以将表皮的结果与用培养中的细胞获得的结果进行比较。 这些研究，以及生物化学调查的相互作用， 受体与其他细胞蛋白质的结合将导致 无细胞系统，其中可以观察到佛波醇酯的作用。 这些研究将有助于描述佛波醇的生物学作用 酯和帮助理解肿瘤促进的生物学过程 和化学致癌作用。