ImmunoAnalysis and Research Specimen Management

核心B将为计划项目提供3个主要功能。首先，Core B将为研究人员提供研究标本，并协助鉴定、免疫染色和图像分析。将提供原始肿瘤或组织微阵列的石蜡包埋切片，用于1)常规H&E确认组织组织学；2)使用ACINUS分析细胞凋亡，使用Ki-67或BrdU结合的mb -1检测细胞增殖，使用标准的单克隆抗体，描述性免疫组织化学和使用视觉评分和/或自动图像分析的定量分析雄激素受体；3)利用免疫组织化学和图像分析鉴定和定量类固醇受体共调节因子、雄激素代谢酶、雄激素调节基因产物（PSA、Nkx3.1、a-微管蛋白、翻译延伸因子la、非神经元烯醇化酶、tomoregulin、硫氧还蛋白还原酶1、Mxi-1和人钾化因子2）、一般转录因子、生长因子及其受体、干细胞和血管生成生物标志物。

Core B wiil provide 3 primary funcfions to the Program Project. First, Core B will provide research specimens to the investigators and assist with identificafion, immunostaining and image analysis. Paraffin-embedded sections will be provided from original tumors or tissue microarrays and used to 1) confirm tissue histology using roufine H&E; 2) analyze apoptosis using ACINUS, proliferation using MIB-1 detecfion of Ki-67 or BrdU incorporation and androgen receptor using monoclonal antibodies by standard, descriptive immunohistochemistry and quanfitative analysis using visual scoring and/or automated image analysis; and 3) idenfify and quanfify steroid receptor co-regulators, androgen metabolism enzymes, androgen-regulated gene products (PSA, Nkx3.1, a-tubulin, translation elongation factor la, non-neuronal enolase, tomoregulin, thioredoxin reducatase 1, Mxi-1 and human kallikrein 2), general transcripfion factors, growth factors and their receptors and stem cell and angiogenesis biomarkers using immunohistochemistry and image analysis. Second, Core B will be responsible for storage, processing and sectioning of clinical (frozen, archival and TMAs) and prostate cancer xenograft (CWR22) research specimens. Core personnel will procure, process and store clinical specimens of castrafion-recurrent prostate cancer obtained from men with advanced prostate cancer who present with urinary retention due to local recurrence. Research specimens will be used to construct tissue microarrays or laser capture microdissected for biochemical measurements of fissue androgens. Lastly, Core B will provide expert biostatistical (Dr. Wilding) and genitourinary pathological service (Dr. Alexiev) to the Program Project. Drs. Mohler and Smith will co-direct the facility and supervise the acfivifies of individuals who already possess high levels of expertise and currently perform these funcfions for the Program Project. Drs. Wilding and Alexiev will lead the efforts to integrate biostatistical and pathological expertise into Core B and the 3 projects. The high level of funcfion of Core B is demonstrated best by Core B's publicafion of 8 methods papers. Core B continues to develop new methods for more accurate measurement of protein expression (4 publicafions) and remains a leader in the use of TMAs for CaP research (2 publications). Core B has performed services that have resulted in manuscripts published, in press and submitted for Project 1 (20), Project 2 (8) and Project 3 (8). Core B also facilitates interaction among the Projects; 12 of 23 publications involved more than 1 Project.