Analysis of a molecular mechanism by which JNK-binding proteins regulate turnover of focal adhesions and formAnalysis of a molecular mechanism by which JNK-binding proteins regulate turnover of focal adhesions and formation of cell polarity

JNK结合蛋白调节粘着斑周转和形成的分子机制分析JNK结合蛋白调节粘着斑周转和细胞极性形成的分子机制分析

基本信息

批准号：
18590287
负责人：
TAKINO Takahisa
金额：
$ 2.57万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590287/
关键词：
Cancer Invasion Cell migration MAPK JNK ERK MT1-MMP ECM 情報伝達インテグリン

项目摘要

Membrane-type 1 matrix metalloproteinase (MT1-MMP) has been implicated in tumor invasion and metastasis. We previously reported that extracellular matrix degradation by MT1-MMP regulates cell migration via modulating sustained integrin-mediated signals. In this study, MT1-MMP-expressing cells were plated onto fibronectin-coated plates and monitored for cell-matrix adhesion formation and fibronectin degradation. The fibronectin was degraded and removed in line with the cell migration track. The migrating cells showed a polarized morphology and were in contact with the edge of fibronectin through the leading edge in which cell-matrix adhesions are concentrated. Expression of MT1-MMP targeted to cell-matrix adhesions by fusing with the focal adhesion targeting (FAT) domain of focal adhesion kinase (FAK) promoted the initial fibronectin lysis at the cell periphery immediately after adhesion. These results suggest that fibronectin is degraded by MT1-MMP located at cell-matrix adhesions which are concentrated at the leading edge of the migrating cells. lb inhibit MT1-MMP at cell-matrix adhesion, the dominant negative form of MT1-MMP (MT1-Pex) was targeted to the cell-matrix adhesion by fusing with the FAT domain (MT1-Pex-FAT). MT1-Pex-FAT accumulated at cell-matrix adhesions and inhibited fibronectin degradation as well as FAK phosphorylation more effectively than parental MT1-Pex. MT1-Pex-FAT was also shown to suppress the invasion of tumor cells into 3-dimensional collagen gel more strongly than MT1-Pex. These results suggest that MT1-MMP-mediated extracellular matrix lysis at cell-matrix adhesions induces the establishment of cell polarity, which facilitates cell-matrix adhesion turnover and subsequent cell migration. This model highlights the role of MT1-MMP at the leading edge of migrating cells.

膜型1基质金属蛋白酶（MT1-MMP）与肿瘤浸润和转移有关。我们先前报道说，MT1-MMP的细胞外基质降解通过调节持续的整联蛋白介导的信号来调节细胞迁移。在这项研究中，将表达MT1-MMP的细胞铺在纤连蛋白涂层的板上，并监测细胞矩阵粘附的形成和纤连蛋白降解。与细胞迁移轨道一致降解纤连蛋白。迁移的细胞表现出极化的形态，并通过浓缩细胞基质粘附的前缘与纤连蛋白的边缘接触。通过与局灶性粘附激酶（FAK）的局灶性靶标（FAT）融合，靶向细胞基质粘附的MT1-MMP表达促进了粘附后的细胞周围的初始纤连蛋白裂解。这些结果表明，纤连蛋白被位于细胞 - 矩阵粘附处的MT1-MMP降解，该粘连集中在迁移细胞的前缘。 LB在细胞矩阵粘附时抑制MT1-MMP，MT1-MMP（MT1-PEX）的主要负形式通过与脂肪结构域（MT1-PEX-FAT）融合来靶向细胞 - 矩阵粘附。比亲属MT1-PEX更有效地在细胞矩阵粘附和抑制纤连蛋白降解以及FAK磷酸化的MT1-PEX-FAT和FAK磷酸化更有效。 MT1-PEX-FAT还显示出比MT1-PEX更强烈地抑制肿瘤细胞进入三维胶原蛋白凝胶的侵袭。这些结果表明，MT1-MMP介导的细胞矩阵粘附处的细胞外基质裂解会诱导细胞极性的建立，从而促进细胞 - 矩阵粘附周转率和随后的细胞迁移。该模型强调了MT1-MMP在迁移细胞前缘的作用。