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: 7345429
• 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 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7345429
• 关键词: Adherens Junction; Androgens; Animals; Area; Armadillo Repeat; Binding; Brain; Cancer Etiology; Cell Line; Cell physiology; Cell-Cell Adhesion; Cell-Matrix Junction; Cells; Co-Immunoprecipitations; Complex; Confocal Microscopy; Cultured Cells; Cytoplasm; DNA-Binding Proteins; Disease; Disruption; Down-Regulation; E-Cadherin; Event; Family; Family Dasypodidae; GTP Binding; Gene Expression; Glutathione S-Transferase; Goals; Growth Factor; Hepatocyte Growth Factor; 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; Protein Overexpression; 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; 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）是美国男性中最常见的恶性肿瘤，也是前列腺癌的第二大病因。 癌症死亡率为了降低显著的发病率和死亡率，预防和治疗CaP的新策略 治疗取决于确定这种疾病中涉及的特定分子机制。 8-连环蛋白是一种独特的含有Armadillo（ARM）结构域的蛋白质，因为它是神经特异性的，并且主要是神经元特异性的。 在大脑中表达。然而，8-连环蛋白表达在前列腺癌中增加， E-cadherin/120 ctn从细胞-细胞连接处脱落，并促进CaP细胞的生长和侵袭。调查这一 在癌症的外周组织中，神经元连环蛋白基因表达的大部分未被探索的区域， 本项目的目的是检验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。这项建议有三个具体目标。 特异性目标1将确定8-连环蛋白如何响应于特异性抗体促进细胞生长/存活和运动性。 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-连环蛋白诱导的信号分子改变。 这些研究将确定8-catenin参与的癌症特异性途径，并将研究它们的作用。 使用蛋白质免疫共沉淀和酵母双杂交分析与S-连环蛋白的相互作用。具体目标 2将确定8-连环蛋白如何与Rho家族小GTP酶相互作用以破坏粘附连接， 促进CaP细胞生长/存活和侵袭。我们将首先确定8-连环蛋白序列负责 这一行动。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/p120 ctn通路之间的相互作用， 揭示了对细胞-细胞粘附和细胞生长/存活和侵袭的可能的差异调节。我们将 还决定了β-连环蛋白的细胞质积累和核信号传导，β-连环蛋白是从E- 通过IQGAP-1检测钙粘蛋白复合物。Specific Aim 3将应用Y311，一种独特的（去磷酸化特异性）单克隆 抗体，和定点诱变，以确定如何8-连环蛋白修饰发生，以及它们是什么。 在CAP中的角色我们还将研究S-连环蛋白的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。最后，我们将产生转基因小鼠， 前列腺组织特异性S-连环蛋白表达，以在动物水平上解决这些问题。 这些研究将把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相关的机器。理解控制8-连环蛋白的翻译后修饰 功能将揭示S-连环蛋白调节基因表达和促进体内CaP的机制。