MOLECULAR PATHOLOGY OF STEM CELLS IN CANCER

描述(申请人的描述)： 经过分子生物学和病理学的培训，目前的目标是 候选人将成为一名独立的内科科学家 癌症的分子病理学。这项提议将解决这一假设 前列腺癌是由异常的干细胞成熟引起的。一种方法 这项测试是为了比较癌症和癌症之间的整体基因表达模式 细胞和干细胞。使用减法等新技术 通过杂交，申请者将尝试分离出 在不同的细胞群体中差异表达。他们会 将这些比较扩展到总基因表达谱的分析 利用基因表达微阵列(GEMS)和/或基因序列分析 随着技术的进步，表达(SAGE)。激光捕获 显微解剖(LCM)可以将单个细胞群体从复杂的细胞中分离出来 混合物。然而，干细胞的分离仍然受到缺乏 合适的记号笔。因此，这些研究的第一部分是针对 分离前列腺干细胞。大鼠的前列腺是尝试 识别前列腺干细胞标记物，因为腺组织可以 在体内反复在退化(细胞凋亡)和再生长之间循环 (增殖)通过操纵血清雄激素。使用此模型， 赞助商和其他人发现端粒酶活性和磷酸化蛋白 PP32是前列腺中的候选干细胞标记物。为了改善 了解前列腺干细胞和组织结构，他们将 端粒酶和pp32表达的细胞定位特征为 与其他候选干细胞标志物和其他 具有良好特征的前列腺细胞类型的标记。申请者将 通过退化、生长的循环监测这些标记的动态 和雄激素恢复。候选的大鼠干细胞标记物将被 在人体内进行评估，以识别相似的细胞类型。假定干细胞 而其他类型的细胞将通过LCM进行可视化和分离。CDNA将会是 为消减杂交和文库制作做准备。成对配对 将使用分离的细胞类型与良性前列腺增生症进行减法杂交， PIN和癌症。申请者的方法是独一无二的，因为 用于分析的材料不是 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和癌症中的成熟，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.