ROLE OF GROWTH FACTORS IN HUMAN PROSTATE CARCINOMA

本建议的总体目标是确定以下机构的作用： 前列腺增生症发病机制中的特异性生长调节途径 癌 选择的生长因子基因和原癌基因的表达 在自然疾病进展期间，将在切除的 患者的标本。 与一个或多个基因相关的基因表达 人类前列腺癌的病理生物学方面将被检查 此外，在人前列腺肿瘤的充分表征的实验模型中， 无胸腺裸鼠的生长和转移。 以下内容的表达 生长因子或生长因子受体基因将在以下方面进行检查： 特异性RNA（通过北方分析）、基因扩增（南方 分析）和蛋白质产物（免疫组织化学和Western分析）， 在可行的情况下：（1）转化生长因子α（TGF-α）及其 受体，（2）表皮生长因子受体（EGFr），（3）c-erbB-2，a 编码修饰的EGF受体的基因，和（4）转化生长因子 β（TGF-β 1）。 免疫组织化学染色与 将使用适当的抗体和蛋白质印迹法（1）将RNA 蛋白质产物表达水平和（2）允许鉴定 合成目标蛋白的细胞类型。 相关表达及 这些基因的功能将通过分析的作用（S） 这些基因产物在肿瘤变异体的生长和/或转移中 人前列腺癌细胞系PC-3和DU 145裸鼠移植瘤模型。 这将伴随着（1）测量特定的基因表达， 现有的PC-3和DU 145变体和（2）选择PC-3变体， 表现出这些基因的高表达或低表达，随后 在裸鼠中进行评价。 在可能的情况下， 这些基因的功能参与将通过遗传学方法来寻找。 通过用适当的转染修饰表型变体 基因。 这些研究将有助于确定这些机构的作用， 生长因子和生长因子受体基因的自然进展 人类前列腺癌。 同样重要的是，这些实验将 允许实验评估这些表达之间的关系， 基因和这种常见人类肿瘤的病理生物学表型。

The overall objective of this proposal is to determine the role(s) of specific growth regulatory pathways in the pathogenesis of human prostatic cancer. Expression of selected growth factor genes and protooncogenes during natural disease progression will be evaluated in surgically-excised specimens from patients. Gene expression which correlates with one or more aspects of the pathobiology of human prostate cancer will be examined further in a well characterized experimental model of human prostatic tumor growth and metastasis in athymic nude mice. Expression of the following growth factors or growth factor receptor genes will be examined in terms of both specific RNA (via Northern analysis), gene amplification (Southern analysis), and protein product (immunohistochemistry and Western analysis), where feasible: (1) transforming growth factor alpha (TGF-alpha) and its receptor, (2) the epidermal growth factor receptor (EGFr), (3) c-erbB-2, a gene coding for a modified EGF receptor, and (4) transforming growth factor beta (TGF-beta1). A combination of immunohistochemical staining with the appropriate antibody and Western blotting will be used (1) to relate RNA levels to protein product expression and (2) to permit identification of the cell type synthesizing the protein of interest. Correlative expression of these genes will be related to function by analysis of the role(s) of these gene products in the growth and/or metastasis of variants of the human prostate carcinoma cell line PC-3 and DU145 in athymic nude mice. This will be accompanied by (1) measurement of specific gene expression in existing PC-3 and DU145 variants and (2) selection of PC-3 variants which exhibit high or low expression of these genes followed by subsequent evaluation in nude mice. Where possible, confirmation of putative functional involvement of these genes will be sought by genetic modification of phenotypic variants via transfection with the appropriate gene(s). These studies will permit identification of the role(s) of these growth factor and growth factor receptor genes in the natural progression of human prostatic cancer. Equally significant, these experiments will permit experimental evaluation of the relation between expression of these genes and the pathobiological phenotype of this common human tumor.