Characterization of prostatic stem cells and prostate cancer-initiating cells

前列腺癌是美国男性癌症相关死亡的第二大原因，我们 使用SCA-1表面抗原增强了前列腺干细胞的活性，并证明了具有 丰富的干细胞活性作为肿瘤启动的靶点，我们的长期目标是充分描述 干细胞和致癌细胞在前列腺癌中的特性。这项研究将为 干细胞生物学和癌症生物学的研究。 我们的具体目标是：(1)全面鉴定前列腺干细胞。(A)我们会调查树干 使用分离的前列腺细胞再生，细胞驻留在特定的前列腺上皮细胞谱系中 系统。我们将(I)分离单个谱系并测试它们的再生能力，以及(Ii)永久标记 并确定其后代的世系状况。这些可以通过创建一个 前列腺基底细胞特异性绿色荧光蛋白标记转基因小鼠模型或通过Creloxp 由前列腺系特异性启动子调控的标记系统。(二)我们会继续筛选 使用表面抗原在前列腺、分裂的前列腺细胞中表达表面抗原并鉴定 利用再生系统获得了具有丰富干细胞活性的组份(S)。(2)确定前列腺癌的起因 细胞。(A)我们将评估基底细胞和腔细胞对致癌的敏感性 转型。我们将通过感染前列腺癌上皮细胞在单个谱系中诱导Pten缺失 表达Cre重组酶的慢病毒PTENIoxp/loxP转基因小鼠模型的建立 特定于世系的启动者。被感染的细胞将在再生系统中进行测试，以确定哪个细胞 血统(S)已经改变(B)我们将确定每个细胞群体的易感性 在Aim1B中由单一致癌刺激或联合致癌刺激诱导恶性转化。细胞分数 来自代表干细胞、短期祖细胞和终末细胞的野生型和p53-/-小鼠 分化的细胞将被慢病毒感染，慢病毒介导不同的致癌信号，如myc， PRb家族蛋白的失活，PTEN-AKT信号通路的扰动等。这个 被感染的细胞将被显微注射到免疫缺陷的宿主小鼠前列腺中，或在再生中进行测试 系统来确定他们引发癌症的能力。

Prostate cancer is the second-leading cause of the cancer-related death in men in the United States, we have enriched prostate stem cell activity using the Sca-1 surface antigen and demonstrated that cells with enriched stem cell activity serve as a target for tumor initiation, our long-term goals are to fully characterize the identity of stem cells and cancer-initiating cells in prostate. This study will provide general insights for the studies on stem cell biology and cancer biology. Our specific aims are:(1) Fully characterize the prostatic stem cells. (A) We will investigate whether stem cells reside in a specific prostatic epithelial cell lineage using the dissociated prostate cell regeneration system. We will (i) isolate individual lineages and test their regenerative capacity, and (ii) permanently mark individual lineages and determine the lineage status of their progeny. These can be achieved by creating a prostate basal cell-specific green fluorescence protein-marked transgenic mouse model or through the Creloxp marking system regulated by prostate lineage-specific promoters. (B) We will continue to screen the expression of surface antigens in prostate, fractionate prostate cells using surface antigens and identify the fraction(s) with enriched stem cell activity using the regeneration system. (2) Identify the prostate cancerinitiating cells. (A) We will evaluate the susceptibility of basal cells and luminal cells to oncogenic transformation. We will induce Pten deletion in individual lineages by infecting prostate epithelial cells from the PTENIoxp/loxp transgenic mouse model with lentivirus that express the Cre recombinase regulated by lineage-specific promoters. Infected cells will be tested in the regeneration system to determine which cell lineage(s) have been transformed (B) We will determine the susceptibility of each cell population fractionated in Aim1 B to malignant transformation induced by single or a combination of oncogenic stimuli. Cell fractions from the wild type and P53-/- mice that represent stem cells, short-term progenitor cells and terminally differentiated cells will be infected with lentivirus that mediate distinct oncogenic signals, such as myc, inactivation of the pRB family proteins, perturbations in the PTEN-AKT signaling pathway and others. The infected cells will be microinjected into immunodeficient host mouse prostate or tested in the regeneration system to determine their capacity to initiate cancer.