ACTIVE RECEPTOR TYROSINE KINASES IN HUMAN PROSTATE CANCER

这个项目的目标是识别和表征生长因子 和生长因子受体的活性，上皮和间充质细胞， 正常前列腺以及前列腺上皮肿瘤/间质成分 原发性和转移性前列腺癌，并确定这些 产生骨种植选择优势的机制 前列腺癌患者的转移。这项建议是根据 前提是正常前列腺上皮细胞和间充质细胞维持 活跃的细胞间信号需要的发展和 这个器官的分化。我们的假设是， 转移性前列腺上皮细胞和间质细胞之间的相互作用 存在正常骨中的元素并模仿正常前列腺的元素， 为肿瘤的生长创造了一个完美的环境。这 这一假设得到了以下事实的支持： 在高达80%死于前列腺癌的患者中， 大多数情况下，他们的临床并发症通常决定 疾病的最终结果。主要目的是翻译基本的 研究结果的临床设置，关于肿瘤的机制 进展 具体目标是： 目的#1：确定发育和临床的作用模式 人前列腺组织中的相关酪氨酸激酶。我们将分析 生长因子受体及其相应生长因子的表型 正常前列腺上皮/间充质细胞上的配体，以及 原发性和转移性前列腺癌的肿瘤/基质成分。 我们将评估一组微孔的敏感性和特异性， 特异性生长因子的表征抗体和分子探针 受体及其相应的生长因子配体，包括EGFr， TGF α、EGF、FGFr、aFGF、bFGF、c-kit、SCF、trk受体和 神经营养素我们将使用免疫组织化学和原位杂交 在连续的组织切片上我们计划补充细胞 通过免疫印迹和北方印迹测定进行定位，使用来自 同一块。将对所有病例进行形态学评价。 目的#2：表征差异表达和活化生长 前列腺肿瘤表达的因子受体/原发性 相对于不同的转移部位（即，淋巴结与骨病变）。 我们将进行高分辨率二维电泳， 通过免疫印迹和微测序技术。这些研究报告将 与Paul Tempst博士（微量化学实验室）合作完成。 目的#3：研究信号转导的机制 通过生长因子与其假定受体的结合介导， 体外模型系统。我们还将分析前列腺癌细胞的共培养物， 癌细胞与来自不同来源的成纤维细胞，包括骨。 将尝试研究转染的NIH 3T3 前列腺特异性生长因子的适当构建体的细胞 癌细胞基于单克隆抗体的阻断试验 特异性生长因子受体也将被引导， 为未来的临床前研究奠定基础。

The goals of this project are to identify and characterize growth factors and growth factor receptors active on epithelial and mesenchymal cells of the normal prostate, as well as on epithelial tumor/stromal elements of primary and metastatic prostate carcinomas, and to determine those mechanisms that produce a selective advantage for the seeding of bone metastases in prostate cancer patients. This proposal is based on the premise that normal prostatic epithelial and mesenchymal cells maintain active intercellular signals required for the development and differentiation of this organ. It is our hypothesis that analogous interactions between metastatic epithelial prostate cells and stromal elements in normal bone are present and mimic those of normal prostate, recreating a perfect environment for tumor seeding and growth. This hypothesis is supported by the fact that osseous metastases are identified in up to 80% of patients dying of prostate cancers, being symptomatic in the majority of cases, and their clinical complications usually dictating the final outcome of the disease. The main objective is to translate basic research findings to the clinical setting, concerning mechanisms of tumor progression. The specific aims are: Aim #1: To determine the modes of action of developmental and clinically relevant tyrosine kinases in human prostatic tissue. We will analyze the phenotype of growth factor receptors and their corresponding growth factor ligands on epithelial/mesenchymal cells of the normal prostate, as well as on tumor/stromal elements of primary and metastatic prostate carcinomas. We will assess the sensitivity and specificity of a panel of well characterized antibodies and molecular probes to specific growth factor receptors and their corresponding growth factor ligands, including EGFr, TGFalpha, EGF, FGFr, aFGF, bFGF, c-kit, SCF, trk receptors and neurotrophins. We will use immunohistochemistry and in situ hybridization on consecutive tissue sections. We plan to complement cellular localization by immunoblotting and northern blot assays, using tissue from the same block. Morphologic evaluation will be assessed in all cases. Aim #2: To characterize the differentially expressed and activated growth factor receptors expressed by prostatic tumor/stromal elements of primary versus different metastatic sites (i.e., lymph node versus bone lesions). We will perform high resolution two-dimensional electrophoresis followed by immunoblotting and microsequencing techniques. These studies will be done in collaboration with Dr. Paul Tempst (Microchemistry Laboratory). Aim #3: To investigate the mechanisms implicated in signal transduction mediated by binding of growth factors to their putative receptors by means of in vitro model systems. We will also analyze co-cultures of prostate cancer cells with fibroblasts from different sources, including bone. Attempts will be made to investigate the action of transfected NIH 3T3 cells with appropriate constructs of specific growth factors on prostate carcinoma cells. Blocking assays based on monoclonal antibodies to specific growth factor receptors will also be conducted and may serve as the basis for future preclinical studies.