A Model System for Human Prostate Tumorigenesis

描述（由申请人提供）：我们假设恶性肿瘤 人前列腺上皮细胞的转化的特征在于 在DNA和RNA水平上获得关键的、最小的遗传改变。 此外，基因表达的这种变化对细胞凋亡的贡献也是显而易见的。 前列腺上皮细胞恶性表型的获得和进展 细胞可以通过成纤维细胞介导的旁分泌相互作用来调节。在 为了验证这一假设，我们提出了一个创新的战略，以创造一个 通过发育上皮细胞系的新生前列腺肿瘤发生模型 与已知的基因改变，然后研究基因型如何驱动 这些细胞中恶性表型的表达。这些研究将定义 人类恶性转化所需的最小遗传事件 前列腺我们还建议检查是否与成纤维细胞 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 完成三个具体目标： 目标1.从人前列腺细胞系中创建模型上皮细胞和成纤维细胞 永生化或永生化和转化的正常前列腺上皮细胞， 和永生化的前列腺成纤维细胞; 目标2.对每个细胞系进行基因型和表型表征，以识别 通过光谱核型分析在DNA水平和通过cDNA在RNA水平的改变 阵列分析，以：a）鉴定与那些 B）在人前列腺肿瘤中观察到的DNA和RNA的相关改变， 恶性表型表达水平; 目标3.通过暴露于条件培养基模拟旁分泌相互作用 成纤维细胞和癌症相关的成纤维细胞，以测试这些 相互作用促进或增强人前列腺恶性肿瘤的表达 上皮细胞表型在体外和体内，并导致改变 利用cDNA阵列在转化的上皮细胞中表达特异性基因 分析. 这些实验将采用新颖的，创新的策略来创造和 验证前列腺模型，以阐明基因型如何驱动表型，以及 在早期恶性转化过程中旁分泌相互作用的影响 和前列腺的进展。该模型的未来应用包括 开发和测试新的化学治疗剂， 前列腺肿瘤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.