BIOLOGICAL AND BIOCHEMICAL ASPECTS OF RAS SIGNALLING

RAS 信号转导的生物学和生化方面

• 批准号: 2023003
• 负责人: DENNIS W STACEY
• 金额: $ 18.37万
• 依托单位: CLEVELAND CLINIC FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-01-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2023003
• 关键词: biological signal transduction; biomarker; cell cycle; cell growth regulation; combination cancer therapy; enzyme linked immunosorbent assay; gel electrophoresis; gene mutation; immunoprecipitation; laboratory rabbit; lipid metabolism; microinjections; molecular genetics; molecular oncology; neutralizing antibody; phospholipids; protein tyrosine kinase; protooncogene

Proto-oncogenes play a critical role in the regulation of cell growth, but it has been difficult to relate the structure of these molecules to their biological functions within the cell.. The goal of this proposal is to determine the role of several critical proto-oncogenes in the control of cell division. In past work a neutralizing antibody to the proto-oncogene ras was identified and microinjected into living cells. The consequences of this treatment upon the proliferative capacity of the cell then indicated the function of ras proteins in normal proliferation. It was clear not only that cellular ras proteins were critical for proliferation, but that ras activity was essential to the function of some but not all retroviral oncogenes. On the basis of these results we have suggested that ras proteins might function to transmit a proliferative signal from one class of proto-oncogenes to another, and that phospholipid metabolism is involved in this signalling. The objective of this proposal is to test these critical hypotheses. First, neutralizing antibodies to other proto-oncogenes believed to function at different stages of the postulated signal transduction pathway will be produced and tested. These antibodies will allow a direct test of the hypothesis that various proto-oncogenes function together in a signal transduction pathway. In addition, previous studies with the anti-ras antibody suggested that phospholipid metabolism might be involved in regulating the biological activity of ras proteins. Recent studies have provided direct biochemical evidence that this might be the case. If these results can be verified and expanded it might provide the first indication of how the activity of cellular ras proteins might be controlled during mitogenesis. The second objective of the proposed study is, therefore, to verify and expand these initial studies. The importance of this work is emphasized by the fact that while cellular ras proteins are believed to play a central role in the control of proliferation, no other indication has yet been reported to explain the biological activation of these proteins. Finally, these microinjection and biochemical studies will be placed in a biological context. The action of oncogenes will be placed in temporal relationship to the known biological marker of cell division . These studies constitute a unique approach in the biological analysis of proliferative genes. the information obtained will undoubtedly aid directly in a molecular understanding of tumor formation since these genes are among those most often mutated in natural human tumors. Furthermore, the biological markers to be tested included antiproliferative agents commonly utilized in anti-tumor therapy.

原癌基因在调节细胞生长中起着关键作用， 但很难将这些分子的结构与 它们在细胞内的生物学功能。 这项提案的目的是 是为了确定几个关键的原癌基因的作用， 控制细胞分裂。 在过去的工作中， 鉴定原癌基因ras并显微注射到活细胞中。 这种处理对细胞增殖能力的影响是不明显的。 细胞则表明ras蛋白在正常增殖中的功能。 很明显，细胞ras蛋白不仅对 增殖，但ras活性是必不可少的功能， 一些但不是全部逆转录病毒致癌基因。 根据这些结果，我们 已经表明ras蛋白可能具有传递 从一类原癌基因到另一类原癌基因的增殖信号，和 磷脂代谢参与了这种信号传导。 本提案的目的是检验这些关键假设。 首先，针对其他原癌基因的中和抗体被认为 在假定的信号转导的不同阶段起作用 将进行生产和测试。 这些抗体将允许 对各种原癌基因功能假说的直接检验 在信号转导通路中结合在一起。 此外，以往的研究 与抗ras抗体的关系表明，磷脂代谢可能 参与调节ras蛋白的生物活性。 最近的研究提供了直接的生物化学证据， 就是这样。 如果这些结果能够得到验证和扩展， 首次表明细胞ras蛋白的活性 可能在有丝分裂过程中受到控制。 的第二个目标 因此，拟议的研究是为了验证和扩大这些初步研究。 这项工作的重要性是强调的事实，虽然细胞 ras蛋白被认为在控制 扩散，尚未报告其他迹象来解释 这些蛋白质的生物活性。 最后，这些微注射 生物化学研究将被置于生物学背景下。 的 癌基因的作用将与已知的 细胞分裂的生物标志物。 这些研究构成了生物学分析的独特方法 增殖基因 所获得的信息无疑将有助于 直接在肿瘤形成的分子理解，因为这些 基因是在自然人类肿瘤中最常突变的基因之一。 此外，待检测的生物标志物包括 抗肿瘤治疗中常用的抗增殖剂。