CORE--AUTOMATED COMBINED IN SITU HYBRIDIZATION AND IMMUNOHISTOCHEMISTRY

核心成分C：自动组合原位杂交和 免疫组织化学(IHC)实验室。 这个核心的目的是为现场自动化提供一种设施 杂交和免疫组织化学，这将特别有助于 成功完成项目1和项目3。使用仪器 在亚利桑那大学开发的Core Leaders已经开发出一种 促进ISH和IHC快速结合的动力学模式自动检测 化验。有了这个程序，反应可以在1-3小时内完成 时间范围(1)。双重标记IHC和ISH的好处是：(A)在联合 定位与发病相关的相互作用分子；(B)在 RNA信息传递到蛋白质产品；(C)展示独特的诊断 免疫构筑特征；及(D)显示肿瘤的克隆性 细胞。该核心将直接影响项目1(具体目标1)，其中 α3和α6整合素表达的重要性问题 将会被检查。它还将有助于AIM 2，其中层粘连蛋白5的作用 关于半桥粒形成的问题将被研究。核心C将支持项目 2在特定目标1和2中，β4和α6T的作用 对侵袭后的转染率进行检测。核心C将通过以下方式支持项目3 为研究表达提供蛋白质和信使核糖核酸的定位 裸鼠皮下和原位LNCaP肿瘤中基质溶素的表达 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.