ANALYSIS OF INSULIN AND MSA ACTION IN CELL HYBRIDS

拟议研究的长期目标是应用这些技术 体细胞遗传学(分离变异细胞系、体细胞 杂交)用于研究胰岛素和胰岛素的作用机制 胰岛素样生长因子。我们之前已经展示过，鼠标 黑色素瘤细胞株PG19对其促生长作用无反应 胰岛素和MSA(一种关系密切的大鼠胰岛素样生长因子 到人类IGF-II)。胰岛素和MSA在PG19黑色素瘤细胞中的作用 似乎在受体后步骤被阻断或解偶联。第一部分 所提议的研究的目的将是确定确切的生化 黑色素瘤细胞对胰岛素的反应被阻断的步骤 在这一点上，杂交种的生长反应发生互补。 我们在这方面的主要工作将包括研究 胰岛素在黑色素瘤和杂交瘤中作用的细胞内介质 细胞。将努力量化参与调查的调解人 对磷酸化和去磷酸化反应均有刺激作用。一个 这些实验的长期目的是识别细胞内 参与胰岛素调节细胞生长的介质。第二 拟议研究的一部分将涉及绘制人类基因图谱 胰岛素、IGF-I和IGF-II受体。在这个项目中，我们将使用 针对受体的单抗以识别 人×鼠体细胞杂交体细胞表面的受体。混合动力车 我们在此项目中使用的克隆面板是通过交叉开发的 人成纤维细胞与小鼠A9细胞株。第三部分的目的 该项目的重点是分离H4-II-E-C3‘大鼠肝癌的变种 对胰岛素的生长反应有缺陷的细胞系。少校 该项目这一部分的重点将是分离出具有 胰岛素作用中的受体后缺陷。该项目与以下方面的相关性 临床医学是，从长远来看，它可能会导致 对胰岛素和胰岛素样生长因子的生化和遗传“途径”的认识 行动。在人类疾病中，这种理解的提高可能会 最终导致改善治疗的是与胰岛素有关的疾病 耐药性，包括一种常见病(II型糖尿病)和一种 以严重胰岛素抵抗为特征的罕见综合征的数量，如 如脂肪萎缩性糖尿病、小妖精和A型胰岛素综合征 抗性和黑棘皮病。

The long term objective of the proposed research is to apply the techniques of somatic cell genetics (isolation of variant cell lines, somatic cell hybridization) to a study of the mechanism of action of insulin and insulin-like growth factors. We have shown previously that the mouse melanoma cell line PG19 is unresponsive to the growth-stimulatory action of insulin and MSA (a rat insulin-like growth factor that is closely related to human IGF-II). Insulin and MSA action in the PG19 melanoma cells appears to be blocked or uncoupled at a postreceptor step. The first part of the proposed research will be aimed at identifying the exact biochemical step at which the response to insulin is blocked in the melanoma cells and at which complementation for the growth response occurs in the hybrids. Our major effort in this regard will involve an examination of intracellular mediators of insulin action in the melanoma and hybrid cells. An effort will be made to quantify mediators involved in the stimulation of both phosphorylation and dephosphorylation reactions. A long term purpose of these experiments is to identify intracellular mediators involved in the regulation of cell growth by insulin. The second part of the proposed research will involve mapping of the human genes for the insulin, IGF-I and IGF-II receptors. In this project, we will use monoclonal antibodies directed against the receptors to identify the receptors on the surface of human x mouse somatic cell hybrids. The hybrid clone panel that we are using for this project was developed by crossing human fibroblasts with the mouse A9 cell line. The aim of the third part of the project will be to isolate variants of the H4-II-E-C3' rat hepatoma cell line having a defective growth response to insulin. The major emphasis of this part of the project will be to isolate variants having postreceptor defects in insulin action. The relevance of this project to clinical medicine is that it may in the long term lead to an improved understanding of the biochemical and genetic "pathway" of insulin and IGF action. Among the human diseases for which this improved understanding may ultimately lead to improved treatment are diseases involving insulin resistance, including a common disease (type II diabetes mellitus), and a number of rare syndromes characterized by severe insulin resistance, such as lipoatrophic diabetes, leprechaunism, and type A syndrome of insulin resistance and acanthosis nigricans.