ACTIVE RECEPTOR TYROSINE KINASES IN HUMAN PROSTATE CANCER

这个项目的目标是确定和描述增长因素。 和生长因子受体激活的上皮细胞和间充质细胞 正常前列腺以及上皮性肿瘤/间质成分 原发癌和转移性前列腺癌，并确定 为骨种植产生选择性优势的机制 前列腺癌患者的转移。这项建议是基于 正常前列腺上皮和间充质细胞维持的前提 发育和发育所需的活跃的细胞间信号 这个器官的分化。我们的假设是，类似的 转移性前列腺上皮细胞与间质的相互作用 正常骨骼中存在的元素与正常前列腺中的元素相似， 为肿瘤的播种和生长创造一个完美的环境。这 确认骨转移的事实支持了这一假说。 在死于前列腺癌的患者中，高达80%的人出现症状 大多数病例及其临床并发症通常决定了 疾病的最终结局。主要目标是将BASIC 关于肿瘤发生机制的临床研究成果 进步。具体目标是： 目标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 原发癌和转移性前列腺癌的肿瘤/间质成分。 我们将评估一组油井的敏感性和特异性 特异性生长因子特异性抗体和分子探针 受体及其相应的生长因子配体，包括EGFR， TGFα、EGF、FGFR、aFGFbFGFc-Kit、SCF、trk受体和 神经营养因子。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Northern印迹分析定位，使用来自 同样的街区。在所有病例中都将进行形态评估。 目的#2：描述差异表达和激活的生长 前列腺癌/原发灶基质细胞因子受体的表达 与不同的转移部位(即，淋巴和骨病变)进行比较。 接下来我们将进行高分辨率的双向电泳法 通过免疫印迹和微测序技术。这些研究将是 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 通过多种途径将生长因子与其可能的受体结合而介导 体外模型系统。我们还将分析前列腺的共培养。 癌细胞与来自不同来源的成纤维细胞，包括骨骼。 将尝试研究转基因NIH3T3的作用 具有适当结构的特定生长因子的前列腺细胞 癌细胞。基于抗病毒单抗的阻断试验 特定的生长因子受体也将被传导，并可能作为 为未来的临床前研究奠定了基础。

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.