Institute of Pathology, RWTH, Aachen University, Aachen, Germany.
J Pathol. 2011 Oct;225(2):172-80. doi: 10.1002/path.2959. Epub 2011 Aug 24.
The location of stem cells in the epithelium of the prostatic acinus remains uncertain, as does the cellular origin of prostatic neoplasia. Here, we apply lineage tracing to visualize the clonal progeny of stem cells in benign and malignant human prostates and understand the clonal architecture of this epithelium. Cells deficient for the mitochondrially-encoded enzyme cytochrome c oxidase (CCO) were identified in 27 frozen prostatectomy specimens using dual colour enzyme histochemistry and individual CCO-normal and -deficient cell areas were laser-capture microdissected. PCR-sequencing of the entire mitochondrial genome (mtDNA) of cells from CCO-deficient areas found to share mtDNA mutations not present in adjacent CCO-normal cells, thus proving a clonal origin. Immunohistochemistry was performed to visualize the three cell lineages normally present in the prostatic epithelium. Entire CCO-deficient acini, and part-deficient acini were found. Deficient patches spanned either basal or luminal cells, but sometimes also both epithelial cell types in normal, hyperplastic or atrophic epithelium, and prostatic intraepithelial neoplasia (PIN). Patches comprising both PIN and invasive cancer were observed. Each cell area within a CCO-deficient patch contained an identical mtDNA mutation, defining the patch as a clonal unit. CCO-deficient patches in benign epithelium contained basal, luminal and endocrine cells, demonstrating multilineage differentiation and therefore the presence of a stem cell. Our results demonstrate that the normal, atrophic, hypertrophic and atypical (PIN) epithelium of human prostate contains stem cell-derived clonal units that actively replenish the epithelium during ageing. These deficient areas usually included the basal compartment indicating the basal layer as the location of the stem cell. Importantly, single clonal units comprised both PIN and invasive cancer, supporting PIN as the pre-invasive lesion for prostate cancer.
前列腺小囊上皮中的干细胞位置尚不确定，前列腺肿瘤的细胞起源也不确定。在这里，我们应用谱系追踪技术来可视化良性和恶性人类前列腺中的干细胞的克隆后代，并了解该上皮的克隆结构。使用双色酶组织化学在 27 个冷冻前列腺切除术标本中鉴定出线粒体编码酶细胞色素 c 氧化酶 (CCO) 缺陷的细胞，并对单个 CCO 正常和缺陷细胞区域进行激光捕获显微切割。发现来自 CCO 缺陷区域的细胞的整个线粒体基因组 (mtDNA) 的 PCR 测序共享在相邻的 CCO 正常细胞中不存在的 mtDNA 突变，从而证明了克隆起源。进行免疫组织化学以可视化通常存在于前列腺上皮中的三个细胞谱系。发现整个 CCO 缺陷小囊以及部分缺陷小囊。缺陷斑块跨越基底或腔细胞，但在正常、增生或萎缩的上皮以及前列腺上皮内瘤变 (PIN) 中，有时也跨越两种上皮细胞类型。观察到包含 PIN 和浸润性癌的斑块。CCO 缺陷斑块内的每个细胞区域都包含相同的 mtDNA 突变，从而将斑块定义为一个克隆单位。良性上皮中的 CCO 缺陷斑块包含基底细胞、腔细胞和内分泌细胞，证明了多谱系分化，因此存在干细胞。我们的结果表明，人类前列腺的正常、萎缩、增生和非典型 (PIN) 上皮含有干细胞衍生的克隆单位，这些单位在衰老过程中积极补充上皮。这些缺陷区域通常包括基底隔室，表明基底层是干细胞的位置。重要的是，单个克隆单位包括 PIN 和浸润性癌，支持 PIN 是前列腺癌的癌前病变。
