FUNCTIONAL ANALYSIS OF THE 9E3/CEF4 GENE

9E3/CEF4 基因的功能分析

• 批准号: 3468961
• 负责人: MANUELA M. MARTINS-GREEN
• 金额: $ 10.47万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 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 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3468961
• 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，其产品是分泌型的 与炎症介质具有很强同源性的蛋白质 进化上保守的。 表达之间存在明显的相关性 这些基因和细胞生长已在培养物中得到证实。 建筑 在这些研究中，我对9E3基因进行了体内研究。 这是 在通过细胞分裂生长的正常组织中表达，不表达 在不通过细胞分裂生长的组织中，并且是短暂的 当细胞离开静息阶段时，在细胞周期早期表达。 9E3 表达在受伤时和伤口愈合过程中增强 （特别是在新生血管形成的区域），并与 肿瘤。 培养物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 观察结果扩展了 gro 基因表达之间的相关性 和生长并表明在自分泌或旁分泌生长中的作用 规定。 此外，受伤时表达升高 伤口因素对表达的高度刺激表明额外的 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可能对于 了解对伤害的反应、伤口愈合、发展 肿瘤和生长调节方面。 为了启动 9E3 基因的功能研究，我开发了一种 原位识别蛋白质的多克隆抗体 从培养的细胞中免疫沉淀它。 我的工作假设是 该基因在伤口愈合和细胞中发挥着重要作用 体内生长。 为了检验这个假设，我建议：（1）纯化和 对 9E3 蛋白进行生化表征； (2) 评估角色的作用 在损伤、伤口愈合和 肿瘤的发展； (3)确定蛋白质是否在细胞中发挥作用 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，在体内具有化学引诱或血管生成特性， 并影响伤口愈合和/或肿瘤生长。