ISOLATION AND CHARACTERIZATION OF PROSTATE STEM CELLS

相当多的证据表明，干细胞存在于成人前列腺上皮的基底细胞区室中。鉴于干细胞的高增殖潜力和肿瘤中未分化细胞的存在，可能的是前列腺异常生长的疾病，包括良性前列腺增生和前列腺癌，衍生自干细胞和/或由干细胞繁殖。尽管如此，前列腺干细胞的特征很差。令人信服的证据使我们假设前列腺干细胞表达前列腺增生抑制蛋白bcl-2。为了验证这一假设，我们的目标是从bcl-2/EGFP转基因小鼠的前列腺中分离荧光基底细胞。我们将确定这个群体中的细胞是否具有干细胞相关的特征。此外，我们将使用诱导型FLP重组酶介导的策略来追踪源自表达bcl-2的细胞的细胞谱系，以确定这些细胞是否是在前列腺中产生更分化的细胞类型的多能祖细胞。虽然这超出了该提议的预算限制，但分离表达bcl-2的细胞将允许对从纯化的基底细胞群制备的cDNA与来自其他细胞群的cDNA进行比较分析。以这种方式，将鉴定在前列腺基底细胞中特异性表达的基因。对特定cDNA序列的进一步分析将导致鉴定在作为干细胞的基底细胞亚群中特异性表达的基因。最终，这项工作将确定前列腺疾病治疗干预的有吸引力的目标。

Considerable evidence suggests that stem cells reside in the basal cell compartment of the epithelium of the adult prostate. Given the high proliferative potential of stem cells and the presence of undifferentiated cells in tumors, it is likely that diseases of abnormal growth of the prostate, including benign prostatic hyperplasia and prostate cancer, deriv. from and/or arc propagated by the stem cells. Despite this, the stem cells of the prostate are poorly characterized. Compelling evidence has led us to hypothesize that prostatic stem cells express the apoptosis-inhibiting protein bcl-2. To test this hypothesis, we aim to isolate fluorescing basal cells from the prostates of bcl-2/EGFP transgenic mice. We shall determine whether cells in this population have characteristics associated with stem cells. In addition, we shall use an inducible FLP recombinase- mediated strategy to track cell lineages deriving from bcl-2 expressing cells to determine whether these Cells arc multipotential progenitor cells that give rise to more differentiated cell types in the prostate. While it is beyond the budgetary constraints of this proposal, isolation of bcl-2 expressing cells will allow for a comparative analysis of cDNA prepared from the purified basal cell population with cDNAs from other cell populations. In this way, genes expressed specifically in prostatic basal cells will be identified. Further analysis of specific cDNA sequences will lead to the identification of genes that are expressed specifically in that subpopulation of basal cells that are stem cells. Ultimately, this work will identify attractive targets for therapeutic interventions for prostatic disease.