MOB-5 IN COLORECTAL & PANCREATIC CANCER

描述：MOB-5在ras介导的RIE-1参与中的作用 细胞转化是这一修订后的新应用的主要焦点。Dr。 梁假设，Mob-5是一种信号分子，位于 致癌RAS。他建议通过以下方式来描述这种基因的功能 鉴定MoB-5相互作用蛋白(S)或受体(S)及其活性 在ras介导的细胞转化中的作用，使用 方法包括逆转录病毒基因转移、反义RNA和 抗体中和。具体目标是1)确定 MOB-5在永生化大鼠肠上皮细胞RAS转化中的作用 (RIE-1)，以及2)确定Mob-5是否在人胰腺中发挥作用 和结直肠癌。第一个目标将通过以下方式实现) 高效表达Mob-5蛋白及制备抗体，b)检测Mob-5表达 在大鼠组织和确定亚细胞位置，c)确定动力学 突变型K-ras基因稳定转染RIE-1细胞对Mob-5表达的影响 细胞，d)逆转录病毒后Mob-5表达的功能特征 RIE-1细胞的介导转化，e)确定MoB-5的作用 反义或Mob-5中和抗体对ras转化的RIE-1细胞的影响 用免疫共沉淀法鉴定与Mob-5相互作用的蛋白质，和g) 确定可能存在Mob-5的细胞表面受体 转化的细胞。具体目标2的研究与建议的研究类似 对于老鼠基因，此外，还评估了一大系列 组织样本和开发基于抗体的临床测试的尝试 癌症患者的血清。

DESCRIPTION: The role of Mob-5 in the participation of ras-mediated RIE-1 cell transformation is the major focus of this revised new application. Dr. Liang hypothesizes that Mob-5 is a signaling molecule downstream of oncogenic ras. He proposes to functionally characterize the gene by identifying Mob-5-interacting protein(s) or receptor(s) and to assess its role in ras-mediated cell transformation, using a combination of methodologies including retroviral gene transfer, antisense RNA, and antibody neutralization. The specific aims are 1) to determine the role of Mob-5 in Ras transformation of immortalized rat intestinal epithelial cells (RIE-1), and 2) to determine whether Mob-5 plays a role in human pancreatic and colorectal cancer. The first aim will be accomplished by a) overexpressing Mob-5 protein and preparing Ab, b) surveying Mob-5 expression in rat tissues and determining subcellular location, c) determining kinetics of Mob-5 expression following stable transfection of mutant K-ras into RIE-1 cells, d) functionally characterizing Mob-5 expression following retrovirus mediated transformation of RIE-1 cells, e) determining the effect of Mob-5 antisense or Mob-5 neutralizing Ab's on ras-transformed RIE-1 cells, f) identifying proteins interacting with Mob-5 by coimmunoprecipitation, and g) determining the presence of a possible cell surface receptor for Mob-5 in transformed cells. Studies in Specific Aim 2 are similar to those proposed for the rat gene with, in addition, an evaluation of a large series of tissue samples and an attempt to develop an Ab based clinical test using serum from cancer patients.