A Model System for Human Prostate Tumorigenesis

描述(由申请人提供)：我们假设 人前列腺上皮细胞转化的特征是 在DNA和RNA水平上获得关键的、最小程度的遗传改变。 此外，基因表达的这种变化对 前列腺上皮细胞恶性表型的获得和进展 细胞可能通过成纤维细胞介导的旁分泌相互作用来调节。在……里面 为了验证这一假设，我们提出了一种创新策略，以创建一个 用培养上皮细胞系建立新生前列腺癌发生模型 已知的基因改变，然后准确地检查基因是如何驱动的 恶性表型在这些细胞中的表达。这些研究将界定 人类恶性转化所需的最小遗传事件 前列腺。我们还建议研究与成纤维细胞之间的联系 通过转化的上皮细胞改变恶性表型的表达， 以及这些旁分泌相互作用如何通过 转化的前列腺上皮细胞。这项工作将通过以下方式完成 完成三个具体目标： 目的1.建立人前列腺上皮细胞和成纤维细胞模型 永生化或永生化并转化为正常前列腺上皮细胞， 永生化的前列腺成纤维细胞； 目的2.鉴定每个细胞系的基因和表型，以确定 DNA光谱核型分析和cDNARNA水平的改变 阵列分析以：a)识别符合以下条件的基因改变 观察到人类前列腺癌，b)DNA和RNA的相关变化 恶性表型表达水平； 目标3.通过暴露于条件培养基来模拟旁分泌相互作用 成纤维细胞和癌症相关成纤维细胞来测试这些 相互作用促进或增强人前列腺癌的恶性表达 在体外和体内的上皮细胞表型，并导致改变 应用基因芯片技术研究转化上皮细胞中特异性基因的表达 分析。 这些实验将应用新颖、创新的策略来创造和 验证前列腺癌模型以阐明基因型如何驱动表型，以及 旁分泌相互作用对早期恶变的影响 和前列腺癌的进展。该模型的未来应用包括 新型化疗药物的开发和试验及新药的开发 前列腺癌发生的新生物标志物。

DESCRIPTION (provided by applicant): We hypothesize that the malignant transformation of human prostate epithelial cells is characterized by the acquisition of key, minimal genetic alterations at the DNA and RNA levels. Furthermore, the contribution of such changes in gene expression to the acquisition and progression of the malignant phenotype by prostate epithelial cells may be modulated through fibroblast-mediated paracrine interactions. In order to test this hypothesis, we propose an innovative strategy to create a model for nascent prostate tumori genesis by developing epithelial cell lines with known genetic alterations, then examining exactly how genotype drives expression of the malignant phenotype in these cells. These studies will define the minimal genetic events required for malignant transformation in the human prostate. We also propose to examine whether association with fibroblast cells alters expression of the malignant phenotype by transformed epithelial cells, and how these paracrine interactions may alter specific gene expression by transformed prostatic epithelial cells. This work will be accomplished through the completion of three specific aims: Aim 1. Create model epithelial and fibroblastic human prostate cell lines from immortalized or immortalized and transformed normal prostate epithelial cells, and immortalized prostate fibroblast cells; Aim 2. Characterize each cell line genotypically and phenotypically to identify alterations at the DNA by spectral karyotyping and at the RNA level by cDNA array analysis to: a) identify genetic alterations consistent with those observed for human prostate tumors, b) correlate alterations at the DNA and RNA levels with expression of the malignant phenotype; Aim 3. Mimic paracrine interactions through exposure to conditioned media from fibroblast and cancer-associated fibroblast cells to test whether these interactions foster or enhance expression of the malignant by human prostate epithelial cells phenotype in vitro and in vivo, and result in the altered expression of specific genes in transformed epithelial cells using cDNA array analysis. These experiments will apply novel, innovative strategies to create and validate a model for prostate to elucidate how genotype drives phenotype, and the influence of paracrine interactions, during early malignant transformation and progression in the prostate. Future applications of this model include the development and testing of new chemotherapeutic agents and the development of novel biomarkers for prostate tumorigenesis.