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申请的主要焦点。 博士 Liang假设Mob-5是一种下游的信号分子， 致癌ras 他建议通过以下方法来描述该基因的功能 鉴定Mob-5相互作用蛋白或受体，并评估其 在ras介导的细胞转化中的作用，使用 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在大鼠肠上皮细胞Ras转化中的作用 （RIE-1），和2）确定Mob-5是否在人胰腺癌中起作用。 和结肠直肠癌。 第一个目标将通过以下方式实现： 过表达Mo B-5蛋白并制备Ab，B）测定Mo B-5表达 在大鼠组织中并确定亚细胞位置，c）确定动力学 突变K-ras稳定转染RIE-1后Mob-5的表达 d）功能性表征逆转录病毒感染后Mob-5的表达， e）确定Mob-5的作用， ras转化的RIE-1细胞上的反义或Mob-5中和Ab，f） 通过免疫共沉淀鉴定与Mob-5相互作用的蛋白质，和g） 确定Mob-5的可能的细胞表面受体的存在， 转化细胞 具体目标2中的研究与提议的研究相似 对于大鼠基因，此外，还评估了大量的 组织样本，并尝试开发一种基于抗体的临床试验， 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.