Aspects of Cadherin/Catenin Complexes

描述：这个项目的长期目标是了解钙粘附素是如何 影响细胞行为的功能。特别是，我们感兴趣的是 N-钙粘附素如何增加人的细胞运动和侵袭行为 上皮性肿瘤细胞。钙粘附素是细胞外基质的跨膜成分 黏附连接是一种细胞间黏附结构，存在于上皮细胞和其他细胞中。 因此，认为黏附蛋白可能参与增加细胞数量的想法 运动性是一个新奇的概念。钙粘附素以组织特异性的方式在 正常组织。我们实验室发现了一种不适当的表达方式 上皮细胞的钙粘附素(在这种情况下，是N-钙粘附素而不是 正常表达的E-钙粘附素)导致细胞运动性增加 以及更多的入侵。 这项建议的具体目标集中在理解 N-钙粘附素在调节细胞运动中起作用。具体来说，我们建议： 1.确定N-钙粘素介导的细胞运动的分子基础。 2.利用转基因小鼠模型研究N-钙粘附素在肿瘤中的作用 进步。 3.确定N-钙粘附素是否通过调节血管生成在血管生成中发挥作用 内皮细胞的运动性。 为了实现这些目标，我们将利用分子构造，通常是 嵌合分子的结合。这些构造将在单元格中表示为 观察它们对细胞行为和表型的影响。蛋白质-蛋白质 互动将通过几种方式确定，其中包括 免疫共沉淀反应，共定位在组织培养细胞和 酵母双杂交分析。我们还将分析蛋白质之间的相互作用。 通过将两种蛋白质的相互作用结构域共转染入真核细胞 然后在完整细胞中共定位(免疫荧光)和 细胞提取物的免疫共沉淀。我们将使用鼠标模型系统来 检测N-钙粘附素在正常乳腺组织中的表达 以确定N-钙粘附素是否只影响细胞行为 肿瘤细胞的角质层。最后，我们将研究N-钙粘附素可能扮演的角色 在正常内皮细胞的运动中发挥作用。 我们预测我们将清楚地识别n-钙粘附素上中介细胞的区域 在不影响细胞黏附的情况下运动。这一知识可能会使这一点成为可能 产生可以用来抑制肿瘤细胞运动的试剂，而不需要 抑制细胞黏附。这种试剂可能有助于防止转移。 肿瘤细胞和/或血管内皮细胞的血管生成。

DESCRIPTION: The long-term goal of this project is to understand how cadherins function to influence cellular behavior. In particular, we are interested in how N-cadherin increases the cell motility and invasive behavior of human epithelial tumor cells. The cadherins are the transmembrane components of the adherens junction, a cell-cell adhesive structure in epithelia and other cells. Thus, the idea that an adhesion protein may be involved in increasing cell motility is a novel one. Cadherins are expressed in a tissue specific manner in normal tissues. Our laboratory discovered that expression of an inappropriate cadherin by an epithelial cell (in this case, N-cadherin rather than the normally expressed E-cadherin) resulted in a cell with increased cell motility and increased invasion. The specific aims of this proposal are focused on understanding the role N-cadherin plays in modulating cell motility. Specifically, we propose to: 1. Determine the molecular basis of N-cadherin-mediated cell motility. 2. Use a transgenic mouse model to investigate the role of N-cadherin in tumor progression. 3. Determine if N-cadherin plays a role in angiogenesis by modulating the motility of endothelial cells. To accomplish these goals, we will make use of molecular constructs, often made up of chimeric molecules. These constructs will be expressed in cells to observe their effect on cellular behavior and phenotype. Protein-protein interactions will be determined by several means which include coimmunoprecipitation reactions, co-localization in tissue culture cells and yeast two hybrid analysis. Protein-protein interactions will also be analyzed by co-transfecting interaction domains of two proteins into eukaryotic cells followed by co-localization in intact cells (immunofluorescence) and co-immunoprecipitation of cell extracts. We will use a mouse model system to examine the effects of expressing N-cadherin in otherwise normal breast epithelial cells to determine if N-cadherin influences cell behavior only in the contex of a tumor cell. Finally, we will examine the role N-cadherin may play in motility of normal endothelial cells. We predict we will clearly identify regions on n-cadherin that mediate cell motility without effecting cell adhesion. This knowledge may make it possible to generate reagents that could be used to inhibit tumor cell motility without inhibiting cell adhesion. Such reagents might be useful to prevent metastasis of tumor cells and/or angiogenesis of endothelial cells.