delta-Catenin and Cell-Cell Adhesion in Prostate Cancer

前列腺癌中的 δ-连环蛋白和细胞间粘附

• 批准号: 7554661
• 负责人: QUN LU
• 金额: $ 18.4万
• 依托单位: EAST CAROLINA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7554661
• 关键词: Adherens Junction; Androgens; Animals; Area; Armadillo Repeat; Binding; Brain; Cancer Etiology; Cell Culture Techniques; Cell Line; Cell physiology; Cell-Cell Adhesion; Cell-Matrix Junction; Cells; Co-Immunoprecipitations; Complex; Confocal Microscopy; Cytoplasm; DNA-Binding Proteins; Disease; Down-Regulation; E-Cadherin; Event; Family; Family Dasypodidae; GTP Binding; Gene Expression; Goals; Growth Factor; Hepatocyte Growth Factor; Human; Intercellular Junctions; Lasers; Lead; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Methods; Modeling; Modification; Molecular; Monoclonal Antibodies; Monomeric GTP-Binding Proteins; Morbidity - disease rate; Neurons; Nuclear; Nuclear Protein; Nuclear Proteins; Pathway interactions; Peptides; Peripheral; Phosphorylation; Phosphotransferases; Post-Translational Protein Processing; Prevention; Prostate; Prostate Adenocarcinoma; Prostatic Neoplasms; Protein Analysis; Protein Array; Proteins; RNA Interference; Research Personnel; Role; Screening procedure; Sequence Homology; Signal Transduction; Signaling Molecule; Site-Directed Mutagenesis; Study Section; Technology; Testing; Tissue Microarray; Tissues; Transgenic Mice; Transgenic Organisms; Yeasts; armadillo proteins; catenin p120ctn protein; cell growth; cell motility; delta-catenin; in vivo; insight; men; mortality; mutant; overexpression; relating to nervous system; response; rho; tumor; tumor xenograft; tumorigenesis; yeast two hybrid system

Prostate cancer (CaP) is the most common malignancy in men in US and the second leading cause of cancer mortality. To reduce the significant morbidity and mortality, new strategies for CaP prevention and treatment depend on the determination of specific molecular mechanisms involved in this disease. 8-Catenin is a unique armadillo (ARM)domain-containing protein in that it is neural specific and primarily expresses in the brain. However, 8-catenin expression increases in prostatic adenocarcinomas, redistributes E-cadherin/120ctn from the cell-cell junction, and promotes CaP cell growth and invasion. To investigate this largely unexplored area of neuronal catenin gene expression in peripheral tissues of cancers, the overall goal of this project is to test the hypothesis that 8-catenin promotes CaP formation and progression by interacting with multiple cellular machineries, including cell-cell and cell-matrix adhesion, cell growth/survival, and invasion. There are three specific aims in this proposal. Specific Aim 1 will determine how 8-catenin promotes cell growth/survival and motility in response to the hepatocyte growth factor and/or androgen using CWR22 tumor xenografts and their derived cell lines. We will broadly screen and profile the S-catenin induced alteration of signaling molecules by array technology. These studies will identify cancer specific pathways that 8-catenin is involved in and will investigate their interactions with S-catenin using protein co-immunoprecipitation and yeast two-hybrid analyses. Specific Aim 2 will determine how 8-catenin interacts with Rho family small GTPases to disrupt adherens junction and promote CaP cell growth/survival and invasion. We will first identify the 8-catenin sequence responsible for this action. Then, the analyses of 8-catenin RNAi or its mutants that are defective in Rac-1 or E-cadherin binding will determine the interactions between GTP-Rac-1/IQGAP-1 and E-cadherin/p120ctn pathways to reveal possible differential modulations on cell-cell adhesion and cell growth/survival and invasion. We will also determine the cytoplasmic accumulation and nuclear signaling of (3-catenin that is released from E- cadherin complexes by IQGAP-1. Specific Aim 3 will apply Y311, a unique (dePhospho-specific) monoclonal antibody, and site-directed mutagenesis to determine how 8-catenin modifications occur and what are their roles in CaP. We will also investigate S-catenin proteolytic fragments and their potentials as DNA binding proteins and nuclear signaling for gene expression. Finally, we will generate transgenic mice displaying prostate tissue specific S-catenin expression to approach these questions at the animal level. These studies will place 8-catenin into the broad context of growth factor and kinase signaling machineries relevant to CaP. Understanding the posttranslational modifications that control 8-catenin functions will unravel mechanisms by which S-catenin modulates gene expression and promotes CaP in vivo.

前列腺癌 (CaP) 是美国男性最常见的恶性肿瘤，也是导致前列腺癌的第二大原因。 癌症死亡率。为了降低显着的发病率和死亡率，CaP 预防和治疗的新策略 治疗取决于该疾病所涉及的特定分子机制的确定。 8-连环蛋白是一种独特的含有犰狳 (ARM) 结构域的蛋白质，因为它具有神经特异性，并且主要 表达在大脑中。然而，8-连环蛋白在前列腺腺癌中表达增加，重新分布 E-cadherin/120ctn 来自细胞与细胞连接处，促进 CaP 细胞生长和侵袭。为了调查此事 癌症周围组织中神经元连环蛋白基因表达的大部分尚未探索的区域，总体 该项目的目标是通过以下方式检验 8-catenin 促进 CaP 形成和进展的假设： 与多种细胞机制相互作用，包括细胞-细胞和细胞-基质粘附、细胞生长/存活、 和入侵。该提案有三个具体目标。 具体目标 1 将确定 8-连环蛋白如何促进细胞生长/存活和运动以响应 使用 CWR22 肿瘤异种移植物及其衍生细胞系的肝细胞生长因子和/或雄激素。我们 将通过阵列技术广泛筛选和分析 S-catenin 诱导的信号分子改变。 这些研究将确定 8-连环蛋白参与的癌症特异性途径，并研究其作用 使用蛋白质免疫共沉淀和酵母双杂交分析与 S-连环蛋白相互作用。具体目标 2 将确定 8-连环蛋白如何与 Rho 家族小 GTPases 相互作用以破坏粘附连接并 促进 CaP 细胞生长/存活和侵袭。我们将首先确定负责的 8-连环蛋白序列 这个动作。然后，分析 Rac-1 或 E-cadherin 缺陷的 8-catenin RNAi 或其突变体 结合将决定 GTP-Rac-1/IQGAP-1 和 E-cadherin/p120ctn 通路之间的相互作用 揭示了细胞间粘附以及细胞生长/存活和侵袭的可能差异调节。我们将 还确定了从 E- 释放的 (3-连环蛋白) 的细胞质积累和核信号传导 IQGAP-1 检测钙粘蛋白复合物。具体目标 3 将应用 Y311，一种独特的（dePhospho 特异性）单克隆抗体 抗体和定点诱变以确定 8-连环蛋白修饰如何发生以及它们的作用是什么 Cap 中的角色。我们还将研究 S-catenin 蛋白水解片段及其作为 DNA 结合的潜力 蛋白质和基因表达的核信号传导。最后，我们将产生表现出 前列腺组织特异性 S-连环蛋白表达可在动物水平上解决这些问题。 这些研究将 8-连环蛋白置于生长因子和激酶信号传导的广泛背景中 与 Cap 相关的机械。了解控制 8-连环蛋白的翻译后修饰 功能将揭示 S-catenin 在体内调节基因表达和促进 CaP 的机制。