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.
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| Begley, Lesa; Monteleon, Christine; Shah, Rajal B et al. (2005) CXCL12 overexpression and secretion by aging fibroblasts enhance human prostate epithelial proliferation in vitro. Aging Cell 4:291-8 |
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