FUNCTIONAL ANALYSIS OF THE 9E3/CEF4 GENE

9E3/CEF4 基因的功能分析

• 批准号: 2185895
• 负责人: MANUELA M. MARTINS-GREEN
• 金额: $ 9.92万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2185895
• 关键词: angiogenesis; biochemistry; cell growth regulation; chemotaxis; gene expression; immunoprecipitation; neoplastic growth; protein purification; protein structure function; wound healing

The 9E3/CEF4 gene is a member of a recently discovered group of genes, sometimes referred to as the gro family, whose products are secreted proteins that have strong homologies to inflammatory mediators and are evolutionarily conserved. A clear correlation between the expression of these genes and cell growth has been demonstrated in culture. Building on these studies, I have investigated the 9E3 gene in vivo. It is expressed in normal tissues that grow by cell division, is not expressed in tissues that do not grow by cell division, and is transiently expressed early in the cell cycle when cells leave the resting stage. 9E3 expression is enhanced upon wounding, during wound healing (especially in areas of neovascularization), and in association with tumors. Cell damage in culture and specific growth factors known to be angiogenic in vivo also stimulate the expression of this gene. My observations extend the correlation between expression of the gro genes and growth and suggest a role in autocrine or paracrine growth regulation. Moreover, the elevated expression upon wounding and the high stimulation of expression by wound factors point to additional roles in wound response and/or angiogenesis. Therefore, discovering the specific function(s) of the products of these genes may be important for the understanding of response to injury, wound healing, development of tumors, and aspects of growth regulation. To initiate functional studies of the 9E3 gene, I have developed a polyclonal antibody that recognizes the protein in situ and immunoprecipitates it from cultured cells. My working hypothesis is that this gene plays an important role in wound healing and in cell growth in vivo. To test this hypothesis, I propose to: (1) Purify and characterize the 9E3 protein biochemically; (2) assess the role of the secreted forms of the protein during injury, wound healing and development of tumors; (3) determine if the protein plays a role in cell growth; (4) determine the kinetics and pathway of protein secretion. These experiments will establish whether or not 9E3 stimulates or represses growth, has chemoattractant or angiogenic properties in vivo, and affects wound healing an/or tumor growth.

9E3/CEF4基因是最近发现的一组基因中的一员， 有时被称为GRO家族，他们的产品是秘密的 与炎症介质有很强同源性的蛋白质 进化上保守的。两种基因表达之间有明显的相关性 这些基因和细胞生长已经在培养中得到证实。建房 在这些研究中，我研究了体内的9E3基因。它是 在通过细胞分裂生长的正常组织中表达，但不表达 在不通过细胞分裂而生长的组织中， 在细胞周期的早期，当细胞离开静止期时表达。 9E3在创伤后、伤口愈合过程中表达增强 (特别是在新生血管领域)，并与 肿瘤。培养中的细胞损伤和已知的特定生长因子 体内血管生成也刺激该基因的表达。我的 观察结果扩大了gro基因表达之间的相关性。 和生长，并暗示在自分泌或旁分泌生长中起作用 监管。此外，伤人时的表情升高和 创伤因子对表达的高刺激指向额外的 在伤口反应和/或血管生成中的作用。因此，发现 这些基因产物的特定功能(S)可能对 对创伤的反应、伤口愈合、发展的理解 肿瘤，以及生长调节的各个方面。 为了启动9E3基因的功能研究，我开发了一种 识别原位蛋白质的多克隆抗体和 免疫沉淀法将其从培养细胞中沉淀出来。我的工作假设是 这种基因在伤口愈合和细胞中起着重要作用 体内生长。为了验证这一假设，我建议：(1)提纯和 对9E3蛋白进行生化表征；(2)评估9E3蛋白的作用 在损伤、伤口愈合和修复过程中蛋白质的分泌形式 肿瘤的发展；(3)确定蛋白质是否在细胞中发挥作用 生长；(4)测定蛋白质分泌的动力学和途径。 这些实验将确定9E3是否刺激或 抑制生长，在体内具有化学吸引或血管生成的特性， 并影响伤口愈合和/或肿瘤生长。