CORE--AUTOMATED COMBINED IN SITU HYBRIDIZATION AND IMMUNOHISTOCHEMISTRY

核心组件C：自动组合原位杂交（ISH）和 免疫组织化学（IHC）实验室。 这个核心的目的是提供一个自动化的设施， 杂交和免疫组织化学，这将特别有助于 项目1和项目3的顺利完成。使用仪器 在亚利桑那大学开发的，核心领导人已经开发了一个 促进快速组合ISH和IHC的动力学模式自动化测定 测定。采用该方法，反应可以在1-3小时内进行， 时间框架（1）。双标记IHC和ISH是有益的：（a）在共- 定位与发病机制相关的相互作用分子;（B）在相关 RNA信息到蛋白质产物;（c）在证明独特的诊断 免疫结构特征;和（d）证明肿瘤的克隆性 细胞该核心将直接影响项目1（具体目标#1），其中 α 3和α 6整联蛋白表达的重要性的问题 将被审查。它也将有助于目标2，其中层粘连蛋白5的作用 半桥粒形成的影响。核心C将支持项目 2在具体目标1和2中，其中β 4和α 6 t 将测试对侵袭的转染。核心C将支持项目3， 提供蛋白质和mRNA定位以研究表达 基质溶解素在皮下和原位LNCaP肿瘤中的模式 免疫缺陷小鼠

CORE COMPONENT C: Automated combined in situ hybridization (ISH) and immunohistochemistry (IHC) laboratory. The purpose of this core is to provide a facility for automated in situ hybridization and immunohistochemistry which will specifically assist in the successful completion of Projects 1 and 3. Using instrumentation developed at the University of Arizona, the Core Leaders have developed a kinetic-mode automated assay which facilitates rapid combined ISH and IHC assays. With this procedure, reactions can be performed within a 1-3 hour time frame (1). Double labeling IHC and ISH are of benefit: (a) in co- localizing interactive molecules relevant to pathogenesis; (b) in relating RNA message to protein product; (c) in demonstrating unique diagnostic immunoarchitectural features; and (d) in demonstrating clonality of tumor cells. This core will directly impact Project 1 (Specific Aim #1) wherein the question of the importance of alpha3 and alpha6 integrin expression will be examined. It will also aid in Aim 2 where the effect of laminin 5 on hemidesmosome formation will be examined. Core C will support Project 2 in Specific Aims 1 and 2 in which the effect of beta 4 and alpha 6t transfection on invasion will be tested. Core C will support Project 3 by providing both protein and mRNA localization to study the expression pattern of matrilysin in subcutaneous and orthotopic LNCaP tumors in immunodeficient mice.