REGULATION OF HUMAN PROSTATE EPITHELIAL CELL GROWTH

描述（由申请人提供）：两种前列腺上皮疾病， 良性前列腺增生（BPH）和前列腺癌，是主要的 美国人面临的健康问题。前列腺上皮失调 细胞增殖是良性和恶性前列腺的中心事件 本申请的目的是确定 胰岛素样生长因子（IGF）和上皮细胞生长 EGF调节人前列腺上皮细胞的生长， 强调IGF受体，EGF受体和 雄激素受体（AR）网络。有待检验的假设是， 雄激素诱导的基因也可被IGF或EGF诱导， 控制前列腺上皮细胞增殖和凋亡的机制。我们 将使用一种新的，充分表征的SV 40 TAg永生化人 前列腺上皮细胞，雄激素受体阴性（M12 AR-）和 雄激素受体阳性（M12 AR+），以实现这些具体目标：（1） 鉴定雄激素受体阳性M12细胞中共同诱导的关键基因 雄激素（DHT）和IGF或EGF的基因表达谱;（2） 确定IGF和EGF调节前列腺细胞生长的机制 在体外和体内。候选基因将过度表达， 通过转染在适当的细胞系中表达降低，或 引入吗啉代反义寡核苷酸或反义构建体。 增殖、凋亡和分化将在体外和体内进行评估。 通过原位注射到无胸腺裸鼠体内;（3）确定 IGF、EGF和雄激素调节基因的表达模式在 目的1对人前列腺良恶性组织进行比较。前列腺组织 将从400例前列腺癌中制备阵列，并通过 免疫组织化学或原位杂交，以确认表达并评估 在我们的实验系统中评估的基因的表达频率。 这些研究将提供全面和独特的见解， 肽生长因子提供替代途径的机制， 控制良性和恶性状态下的前列腺上皮细胞增殖。

DESCRIPTION (provided by applicant): Two diseases of the prostatic epithelium, benign prostatic hyperplasia (BPH) and prostate cancer, are among the major health problems faced by American men. Deregulation of prostatic epithelial cell proliferation is a central event in both benign and malignant prostatic disease about The objective of this application is to determine the mechanism(s) by which insulin-like growth factor (IGF) and epithelial growth factor (EGF) regulate the growth of human prostate epithelial cells, with emphasis on interactions between the IGF receptor, EGF receptor and the androgen receptor (AR) networks. The hypothesis to be tested is that androgen-induced genes also inducible by IGF or EGF provide an alternate mechanism for control of prostate epithelial cell proliferation and behavor. We will use a novel, well characterized family of SV40TAg immortalized human prostate epithelial cells, both androgen receptor negative (M12AR-) and androgen receptor positive (M12AR+) to achieve these specific aims: (1) To identify key genes induced in common in androgen receptor positive Ml2 cells by both androgen (DHT) and IGF or EGF by applying cDNA microarray analysis; (2) To determine the mechanism(s) by which IGF and EGF regulate prostate cell growth in vitro and in vivo. Candidate genes will be either over expressed or reduced in expression in the appropriate cell line by transfection or introduction of morpholino antisense oligonucleotides or antisense constructs. Proliferation, apoptosis, and differentiation will be assessed in vitro and in vivo by orthotopic injection into athymic nude mice; (3) To determine the expression patterns of the IGF, EGF, and androgen regulated genes identified in Aim 1 among human benign and malignant prostatic tissues. Prostate tissue arrays will be prepared from 400 prostate cancers and screened by immunohistochemistry or in situ hybridization, to confirm expression and assess the frequency of expression of the genes evaluated in our experimental system. These studies will provide comprehensive and unique insights into the mechanisms by which peptide growth factors provide alternate pathways to control prostate epithelial cell proliferation in benign and malignant states.