MECHANISMS OF PROSTATE DEVELOPMENT

前列腺发育机制

• 批准号: 3248732
• 负责人: RAJVIR DAHIYA
• 金额: $ 18.24万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-30 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3248732
• 关键词: RNase protection assay; androgens; athymic mouse; benign prostate hyperplasia; cell cell interaction; cell differentiation; cell growth regulation; epithelium; growth factor; growth factor receptors; hormone regulation /control mechanism; human fetus tissue; immunocytochemistry; mesenchyme; northern blottings; paracrine; polymerase chain reaction; prostate; smooth muscle; transforming growth factors; western blottings

The MAIN GOAL of this project is to understand the mechanisms regulating cellular growth of human fetal and adult prostate with the emphasis on the role of mesenchyme-epithelial interactions and growth factors (KGF, TGFalpha, and TGFbeta). The RATIONALE for this proposal is that the developing prostate is an appropriate paradigm for mechanistic studies of growth regulation by paracrine mechanisms in normal and hyperplastic prostates. This project is based on the HYPOTHESIS that the endocrine and developmental control of prostatic cell growth may be regulated by cell-cell interactions mediated by the local production and action of peptide growth factors. In order to test this hypothesis that paracrine mechanisms may be involved in growth and differentiation of developing and adult prostate, we will pursue the following SPECIFIC AIMS: 1) To elucidate the paracrine mechanisms in the regulation of human prostatic growth and differentiation. This will be achieved by grafting human fetal prostates into male athymic nude mice. The resultant growing human fetal prostatic tissue will be examined for a) growth kinetics (doubling time and 3H-thymidine labeling index), ductal elongation and branching morphogenesis, and expression of differentiation markers (PSA, PSAP) by immunohistochemistry and western blot; b) expression of KGF, TGFalpha, and TGFbeta and their receptors by RT-PCR, nuclease protection assay, northern blot, and immunohistochemistry; c) the effects of exogenous KGF, TGFalpha, and TGFbeta and their neutralizing antibodies on organ cultures of growing human fetal vs. adult human prostate (BPH) tissue; d) regulation of KGF, TGFalpha, TBFbeta levels by androgens; e) determination of temporal and spatial aspects of growth factors and their receptors in fetal and BPH tissue. 2) To determine the growth promoting and morphogenetic effects of human urogenital sinus mesenchyme (UGM) on growth-quiescent adult human prostatic epithelium. For this purpose we will isolate UGM and combine it with human prostatic epithelial cells. The resultant tissue recombinant will be grown in athymic nude mice and will be characterized for growth and development as described under Specific Aim #1. 3) To investigate the role of growth factors and cell- cell interactions in the prostate using a new BPH-derived epithelial cell line. This specific aim will be achieved by determining KGF, TGFalpha, and TGFbeta as regulators of prostatic epithelial growth and development. We will also examine the role of prostatic epithelium in the induction of smooth muscle differentiation. Accomplishment of these objectives will elucidate the mechanisms regulating prostatic growth and development by mesenchymal epithelial interaction mediated by the local production and action of growth factors. Through this multidisciplinary approach, we will definitely establish the role of these growth factors as paracrine/autocrine growth regulators in the human prostatic development. The LONG-TERM OBJECTIVES of the proposal are to develop the best strategy for regulation of prostatic growth.

该项目的主要目标是了解调节的机制 人类胎儿和成人前列腺的细胞生长，重点 间质上皮相互作用和生长因子的作用（kgf， TGFALPHA和TGFBETA）。 该提议的理由是 发展前列腺是机械研究的合适范式 正常和增生的旁分泌机制调节生长调节 前列物。 该项目基于内分泌的假设 前列腺细胞生长的发展控制可能受 细胞电池相互作用由局部生产和动作介导 肽生长因子。 为了检验旁分泌的假设 机制可能参与了发展的生长和差异化 和成人前列腺，我们将追求以下特定目标：1） 阐明人类前列腺调节中的旁分泌机制 生长与分化。 这将通过嫁接人来实现 胎儿前列腺裸鼠。 由此产生的人类 将检查胎儿前列腺组织的a）生长动力学（加倍 时间和3H-胸腺苷标记指数），导管伸长和分支 形态发生和分化标记的表达（PSA，PSAP） 免疫组织化学和蛋白质印迹； b）kgf的表达，tgfalpha， 通过RT-PCR，核酸酶保护测定法，TGFBETA及其受体， 北印迹和免疫组织化学； c）外源性kgf的影响， TGFALPHA和TGFBETA及其在器官培养的中和抗体 人类胎儿与成年人前列腺（BPH）组织的生长； d） 雄激素对KGF，TGFALPHA，TBFBETA水平的调节； e） 确定生长因素的时间和空间方面 胎儿和BPH组织中的受体。 2）确定促进增长 人类尿量窦间充质（UGM）对形态发生作用对 成年人前列腺上皮。 为此，我们 将分离UGM并将其与人类前列腺上皮细胞结合使用。 所得的组织重组将在无胸腺裸鼠中生长， 如下所述，将以生长和发展为特征 特定目标＃1。 3）研究生长因子和细胞的作用 使用新的BPH衍生的上皮细胞中前列腺中的细胞相互作用 线。 通过确定kgf，tgfalpha，将实现这一特定目标 TGFBETA是前列腺上皮生长和发育的调节者。 我们还将检查前列腺上皮在诱导中的作用 平滑肌差异。 实现这些目标 将阐明调节前列腺增长和发展的机制 通过局部生产介导的间质上皮相互作用 和生长因素的作用。 通过这种多学科方法， 我们一定会确定这些增长因素的作用 人类前列腺发展中的旁分泌/自分泌生长调节剂。 该提案的长期目标是制定最佳策略 用于调节前列腺增长。