Planz B, Wang Q, Kirley S D, Marberger M, McDougal W S
Department of Urology, University of Vienna, Vienna, Austria.
J Urol. 2001 Aug;166(2):678-83.
Stromal-epithelial interactions of growth factors and the androgen receptor may have implications for the pathophysiology of benign and neoplastic transformation of the human adult prostate. We investigated a possible interaction of keratinocyte growth factor with its receptor as well as with the androgen receptor signaling pathway in human prostatic epithelial cells.
Human prostatic epithelial cells were obtained from explant primary culture, established in DU145 cell conditioned medium and maintained in keratinocyte serum-free medium with supplements. Epithelial cells were characterized by light and electron microscopy, and immunocytochemical study using epithelial and mesenchymal markers. Androgen receptor, keratinocyte growth factor receptor and keratinocyte growth factor messenger RNA expression was measured by polymerase chain reaction (PCR). The response to 0.01 to 10 nM. dihydrotestosterone, 10 microM. flutamide and 1 to 1,000 ng./ml. keratinocyte growth factor was tested by [3H] thymidine assay. The difference in keratinocyte growth factor receptor and androgen receptor gene expression after treatment with and without keratinocyte growth factor and flutamide were determined by quantitative multiplex PCR and quantitated using densitometry analysis.
Immunocytochemical and electron microscopy characterization revealed typical epithelial differentiation. PCR showed keratinocyte growth factor receptor and androgen receptor expression in epithelial cultured cells but no keratinocyte growth factor expression. Epithelial cells showed a significant time and dose dependent stimulation of cell proliferation with keratinocyte growth factor and dihydrotestosterone (p <0.05). When combined with the anti-androgen flutamide the effect of 100 ng./ml. keratinocyte growth factor was significantly decreased (p <0.05). At 100 ng./ml. keratinocyte growth factor quantitative multiplex PCR revealed stimulated keratinocyte growth factor receptor and androgen receptor messenger RNA expression.
These results show that keratinocyte growth factor up-regulates the keratinocyte growth factor and androgen receptors in the absence of androgen. Thus, the androgen signaling pathway may be activated by growth factors such as keratinocyte growth factor in an androgen deficient environment.
生长因子与雄激素受体的基质-上皮相互作用可能对人类成年前列腺的良性和肿瘤性转化的病理生理学具有重要意义。我们研究了角质形成细胞生长因子与其受体以及在人前列腺上皮细胞中与雄激素受体信号通路之间可能存在的相互作用。
从外植体原代培养中获取人前列腺上皮细胞，在DU145细胞条件培养基中建立培养体系，并在添加了补充剂的无角质形成细胞血清培养基中维持培养。通过光镜和电镜以及使用上皮和间充质标志物的免疫细胞化学研究对上皮细胞进行表征。通过聚合酶链反应（PCR）测量雄激素受体、角质形成细胞生长因子受体和角质形成细胞生长因子信使核糖核酸的表达。通过[3H]胸苷测定法检测对0.01至10 nM.二氢睾酮、10 microM.氟他胺和1至1000 ng./ml.角质形成细胞生长因子的反应。通过定量多重PCR确定在有和无角质形成细胞生长因子及氟他胺处理后角质形成细胞生长因子受体和雄激素受体基因表达的差异，并使用密度测定分析进行定量。
免疫细胞化学和电镜表征显示典型的上皮分化。PCR显示上皮培养细胞中有角质形成细胞生长因子受体和雄激素受体表达，但无角质形成细胞生长因子表达。上皮细胞显示出角质形成细胞生长因子和二氢睾酮对细胞增殖有显著的时间和剂量依赖性刺激（p<0.05）。当与抗雄激素氟他胺联合使用时，100 ng./ml.角质形成细胞生长因子的作用显著降低（p<0.05）。在100 ng./ml.角质形成细胞生长因子时，定量多重PCR显示角质形成细胞生长因子受体和雄激素受体信使核糖核酸表达受到刺激。
这些结果表明，在无雄激素的情况下，角质形成细胞生长因子上调角质形成细胞生长因子和雄激素受体。因此，在雄激素缺乏的环境中，雄激素信号通路可能被角质形成细胞生长因子等生长因子激活。
