Adhesion Signaling and Tumor Cell Progression

粘附信号传导和肿瘤细胞进展

• 批准号: 6300589
• 负责人: J THOMAS PARSONS
• 金额: $ 13.25万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-08 至 2001-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6300589
• 关键词: athymic mouse; biological signal transduction; cell adhesion; cell line; cell motility; extracellular matrix; extracellular matrix proteins; flow cytometry; focal adhesion kinase; growth factor receptors; integrins; male; metastasis; mitogen activated protein kinase; neoplastic cell; neoplastic process; paxillin; phosphorylation; prostate neoplasms; protein tyrosine kinase; receptor expression

The long range goal of this Project is to understand how alterations in cellular environment, particularly change in the interactions of cells with the extracellular matrix (ECM) and growth factors with their receptors, influencing the progression and metastatic behavior of prostate tumor cells. We probe to investigate the changes in adhesion signaling pathways using a well characterized panel of prostate cell lines, representing different stages of progression along a pathway from poorly tumorigenic to highly tumorigenic and metastatic. We will focus on key components of the integrin signaling pathway: the protein tyrosine kinases. Focal Adhesion Kinase (FAK) and Src; a putative downstream effector of FAK/Src function, paxillin (a focal adhesion protein); downstream effectors of integrin signaling, MAP kinase; and putative integrin regulated genes. We will also investigate the role of growth factor mediated signaling pathways as co-regulators of integrin signaling, focusing on the possible synergistic role of growth factors in modulating adhesion signaling through the integrin pathway. The specific aims are three-fold: 1) Using a panel of prostate cancer cell lines we will compare the repertoire of integrin receptors and determine their functional binding to ECM proteins; characterize the integrin-specific signaling responses in the different cell lines by assessing the activation of FAK, the tyrosine phosphorylation of paxillin, and the activation of MEK/MAP kinase; compare the repertoire of growth factor receptors expressed in these cells, and establish biological parameters of adhesion signaling by comparing the motility and tumorigenicity of individual cell lines. 2) Using the poorly tumorigenic cell line LNCaP as a recipient cell, we will determine whether the exogenous activation of integrin signaling pathways significantly enhances "progression" of cells to a more metastatic cell population. 3) Using the tumorigenic cell lines, C4-2, derived from LNCaP, as well as the unrelated tumorigenic cell lines, PC3, we will determine if inhibitor of the integrin signaling pathway using specific dominant negative )dn) expression constructs, block tumorigenicity and/or metastasis. These experiment seek to verify the importance of the adhesion signaling pathways in the development and progression of prostate cancer.

本项目的长期目标是了解细胞环境的改变，特别是细胞与细胞外基质（ECM）和生长因子与其受体相互作用的变化，如何影响前列腺肿瘤细胞的进展和转移行为。我们使用一组特征良好的前列腺细胞系来研究粘附信号通路的变化，这些细胞系代表了从低致瘤性到高致瘤性和转移性通路的不同进展阶段。我们将集中在整合素信号通路的关键组成部分：蛋白酪氨酸激酶。粘着斑激酶（FAK）和Src; FAK/Src功能的推定下游效应物桩蛋白（paxillin）（粘着斑蛋白）;整联蛋白信号传导的下游效应物MAP激酶;和推定整联蛋白调节基因。我们还将研究生长因子介导的信号传导途径作为整合素信号传导的共同调节因子的作用，重点关注生长因子通过整合素途径调节粘附信号传导的可能协同作用。具体目标有三个：1）使用一组前列腺癌细胞系，我们将比较整联蛋白受体的库并确定它们与ECM蛋白的功能结合;通过评估FAK的活化、桩蛋白的酪氨酸磷酸化和MEK/MAP激酶的活化来表征不同细胞系中的整联蛋白特异性信号传导应答;比较这些细胞中表达的生长因子受体的库，并通过比较单个细胞系的运动性和致瘤性来建立粘附信号传导的生物学参数。2)使用致瘤性差的细胞系LNCaP作为受体细胞，我们将确定整合素信号通路的外源性激活是否显著增强细胞向更具转移性的细胞群的“进展”。3)使用衍生自LNCaP的致瘤细胞系C4-2以及不相关的致瘤细胞系PC 3，我们将确定使用特异性显性失活（dn）表达构建体的整联蛋白信号传导途径的抑制剂是否阻断致瘤性和/或转移。这些实验旨在验证粘附信号通路在前列腺癌发生和发展中的重要性。