Department of Urology, University Hospital Tuebingen, Tuebingen, Germany.
Prostate. 2013 Oct;73(14):1576-90. doi: 10.1002/pros.22708. Epub 2013 Jul 8.
Benign prostatic hyperplasia (BPH) and prostate cancer (PCa) are common abnormalities in elderly men. It is considered that epithelial stem cells are involved in the etiology and development of both diseases. To distinguish aberrant from normal cells, the knowledge about primary epithelial stem/progenitor cells (ES/P) is essential. The aim of this study was to examine the role of surface markers to distinguish between different subsets of prostate basal epithelium.
The expression pattern of prostate tissue single cell suspensions was analyzed by flow cytometry using different markers. Sorted cell populations were examined for their clonogenic capacity and the resulted colonies were analyzed with flow cytometry, Western blot, and qPCR for stem cell, basal, and luminal epithelium markers. Additionally, the histological localization of the examined markers was determined using immunofluorescence.
Using the combination of CD49f, Trop-2, and surface CD24, basal cell subsets with distinct differentiation capacities were dissected (CD49f(+) Trop-2(+) CD24(-) and CD49f(+) Trop-2(+) CD24(+) ). Although cells from the two subsets gave rise to similar basal colonies, qPCR of primary tissue revealed that higher levels of basal marker expression were detected in the CD49f(+) Trop-2(+) CD24(-) subset. Immunofluorescence analysis showed a prominent expression of CD24 by luminal and basal cells.
Subsets with distinct differentiation capacities within the basal epithelium (CD49f(+) Trop-2(+) CD24(-) and CD49f(+) Trop-2(+) CD24(+) ) can be distinguished in human prostate. CD24 is a marker expressed on the basal transit-amplifying cells (transition cells) and may play a role in the differentiation and migration of ES/P cells to the luminal layer. The knowledge of this mechanism is of relevance for treatment of both diseases.
良性前列腺增生(BPH)和前列腺癌(PCa)是老年男性常见的异常。人们认为上皮干细胞参与了这两种疾病的病因和发展。为了区分异常细胞和正常细胞,了解原发性上皮干细胞/祖细胞(ES/P)是必不可少的。本研究旨在探讨表面标志物在区分前列腺基底上皮不同亚群中的作用。
使用不同的标志物通过流式细胞术分析前列腺组织单细胞悬液的表达模式。对分选的细胞群体进行克隆形成能力检测,并对形成的集落进行流式细胞术、Western blot 和 qPCR 分析,以检测干细胞、基底和腔上皮标志物。此外,还使用免疫荧光法确定了所检查标志物的组织学定位。
使用 CD49f、Trop-2 和表面 CD24 的组合,可分离出具有不同分化能力的基底细胞亚群(CD49f(+) Trop-2(+) CD24(-) 和 CD49f(+) Trop-2(+) CD24(+) )。尽管两个亚群的细胞都产生了类似的基底集落,但对原组织进行 qPCR 分析显示,CD49f(+) Trop-2(+) CD24(-) 亚群中检测到更高水平的基底标志物表达。免疫荧光分析显示 CD24 在腔和基底细胞中均有明显表达。
可以在人前列腺中区分具有不同分化能力的基底上皮亚群(CD49f(+) Trop-2(+) CD24(-) 和 CD49f(+) Trop-2(+) CD24(+) )。CD24 是基底迁移扩增细胞(过渡细胞)上的标志物,可能在 ES/P 细胞向腔层分化和迁移中发挥作用。了解这种机制对于这两种疾病的治疗具有重要意义。