FUNCTIONAL ANALYSIS OF THE 9E3/CEF4 GENE

9E3/CEF4 基因的功能分析

• 批准号: 3468962
• 负责人: MANUELA M. MARTINS-GREEN
• 金额: $ 10.36万
• 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/3468962
• 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的成员 有时被称为GRO家族，其产品被分泌 对炎症介质具有强大同源的蛋白质，并且是 进化保守。 表达之间的明显相关性 这些基因和细胞生长已在培养中得到证明。 建筑 在这些研究中，我研究了体内的9E3基因。 这是 在按细胞分裂生长的正常组织中表达，未表达 在不按细胞分裂生长的组织中，并且是瞬时的 当细胞离开静止阶段时，在细胞周期的早期表达。 伤口愈合期间受伤后9E3表达增强 （特别是在新血管形成的地区），并与 肿瘤。 培养物中的细胞损伤和已知的特定生长因子 血管生成体内还刺激该基因的表达。 我的 观察扩展了GRO基因表达之间的相关性 和增长并暗示在自分泌或旁分泌生长中发挥作用 规定。 此外，受伤和 伤口因子对表达的高刺激指向额外 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对于 理解对伤害，伤口愈合的反应，发展 肿瘤和生长调节的各个方面。 为了启动9E3基因的功能研究，我已经开发了一个 识别原位蛋白质的多克隆抗体和 免疫沉淀从培养的细胞中。 我的工作假设是 该基因在伤口愈合和细胞中起着重要作用 体内生长。 为了检验这一假设，我建议：（1）净化和 以生化为特征9E3蛋白； （2）评估 受伤期间蛋白质的分泌形式，伤口愈合和 肿瘤的发展； （3）确定蛋白质是否在细胞中起作用 生长; （4）确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，在体内具有趋化因子或血管生成特性， 并影响伤口愈合AN/或肿瘤生长。