MOLECULAR PATHOLOGY OF STEM CELLS IN CANCER

描述（申请人描述）： 通过分子生物学和病理学的培训， 候选人将成为一名独立的医生科学家， 癌症的分子病理学 本提案将解决以下假设： 前列腺癌是由异常的干细胞成熟引起的 的一种方法 这项测试是为了比较癌症之间的总体基因表达模式， 细胞和干细胞。 使用新的技术，如减法 为了进行杂交，申请人将尝试分离 在不同的细胞群体中差异表达。 他们将 将这些比较扩展到总基因表达谱的分析 使用基因表达微阵列（GEMS）和/或基因序列分析 表达（SAGE）随着技术的进步。 激光捕获 显微切割（LCM）可以从复杂的细胞中分离出单个细胞群。 混合物。 然而，干细胞的分离仍然受到缺乏 合适的标记。 因此，这些研究的第一部分是针对 分离前列腺干细胞 老鼠的前列腺是理想的尝试 识别前列腺干细胞标志物，因为腺组织可以 在体内在退化（凋亡）和再生长之间反复循环 （增殖）通过操纵血清雄激素。 使用该模型， Sponsor和其他人发现端粒酶活性和磷蛋白 PP 32是前列腺中的候选干细胞标志物。 提高 了解前列腺干细胞和组织组织，他们将 将端粒酶和pp 32表达的细胞定位表征为 与其他候选干细胞标志物和其他 前列腺细胞类型的充分表征的标志物。 申请人会 通过退化、生长、 和雄激素恢复。 候选大鼠干细胞标志物将被 在人体中进行评估以识别相似的细胞类型。 干细胞 其他细胞类型将通过LCM可视化和分离。 cDNA将 制备用于消减杂交和文库产生。 成对 将使用分离的BPH细胞类型进行消减杂交， PIN和癌。 申请人的方法是独特的，因为开始 用于分析的材料不是细胞内细胞类型的复杂混合物。 前列腺 这些研究应该会对改变的干细胞产生新的见解 在BPH、PIN和癌中的成熟，并可能提供新的分子靶点 用于前列腺癌的诊断、预后和治疗。

DESCRIPTION (Applicant's Description): With training in molecular biology and pathology, the current goal of the candidate is to become an independent physician scientist investigating the molecular pathology of cancer. This proposal will address the hypothesis that prostate cancer arises from aberrant stem cell maturation. One way to test this is to compare overall gene expression patterns between cancer cells and stem cells. Using new techniques such as subtraction hybridization, the applicants will attempt to isolate genes that are differentially expressed in the different cell populations. They will extend these comparisons to the analysis of total gene expression profiles using Gene Expression Micro-arrays (GEMS) and/or Serial Analysis of Gene Expression (SAGE) as the technology advances. Laser Capture Micro-dissection (LCM) can isolate individual cell populations from complex mixtures. Yet, isolation of stem cells is still hindered by the lack of suitable markers. Thus, the first portion of these studies is directed at isolating prostate stem cells. The rat prostate is ideal for attempts to identify prostate stem cell markers since the glandular tissue can be repeatedly cycled in vivo between involution (apoptosis) and re-growth (proliferation) by manipulating serum androgens. Using this model, the sponsor and others found that telomerase activity and the phospho-protein pp32 are candidate stem cell markers in the prostate. To improve the understanding of prostate stem cells and tissue organization, they will characterize the cellular localization of telomerase and pp32 expression as compared with that of other candidate stem cell markers and other well-characterized markers of prostate cell types. The applicants will monitor the dynamics of these markers through cycles of involution, growth and androgen restoration. Candidate rat stem cell markers will then be evaluated in the human to identify similar cell types. Putative stem cell and other cell types will be visualized and isolated by LCM. cDNA will be prepared for subtractive hybridization and library production. Pair-wise subtractive hybridizations will be made using isolated cell types with BPH, PIN, and carcinoma. The applicants' approach is unique in that the starting material for analysis is not a complex mixture of cell types within the prostate. These studies should produce new insights into altered stem cell maturation in BPH, PIN and carcinoma, and may provide new molecular targets for diagnosis, prognosis and treatment of prostate cancer.