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细胞室。鉴于干细胞的高增殖能力和肿瘤中未分化细胞的存在，前列腺异常生长的疾病，包括良性前列腺增生症和前列腺癌，很可能衍生出。来自干细胞和/或由干细胞繁殖。尽管如此，前列腺干细胞的特征并不理想。令人信服的证据引导我们假设，前列腺干细胞表达凋亡抑制蛋白bcl2。为了验证这一假设，我们的目标是从bcl2/EGFP转基因小鼠的前列腺中分离出发出荧光的基底细胞。我们将确定这一群体中的细胞是否具有与干细胞相关的特征。此外，我们将使用可诱导的FLP重组酶介导的策略来跟踪来自bcl2表达细胞的细胞谱系，以确定这些细胞是否是在前列腺中产生更多分化细胞类型的多潜能前体细胞。虽然这超出了这一建议的预算限制，但分离表达bcl2的细胞将使从纯化的基底细胞群中制备的cDNA与从其他细胞群中获得的cDNA进行比较分析。通过这种方式，将识别在前列腺基底细胞中特异表达的基因。对特定的cdna序列的进一步分析将导致识别在作为干细胞的基底细胞亚群中特异表达的基因。最终，这项工作将为前列腺癌的治疗干预确定有吸引力的目标。

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.