Pascal Laura E, Oudes Asa J, Petersen Timothy W, Goo Young Ah, Walashek Laura S, True Lawrence D, Liu Alvin Y
Department of Urology, and the Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98195, USA.
BMC Urol. 2007 Apr 10;7:6. doi: 10.1186/1471-2490-7-6.
Identification and characterization of the prostate stem cell is important for understanding normal prostate development and carcinogenesis. The flow cytometry-based side population (SP) technique has been developed to isolate putative adult stem cells in several human tissue types including the prostate. This phenotype is mainly mediated by the ATP-binding cassette membrane transporter ABCG2.
Immunolocalization of ABCG2 was performed on normal prostate tissue obtained from radical prostatectomies. Normal human prostate SP cells and ABCG2+ cells were isolated and gene expression was determined with DNA array analysis and RT-PCR. Endothelial cells were removed by pre-sorting with CD31.
ABCG2 positive cells were localized to the prostate basal epithelium and endothelium. ABCG2+ cells in the basal epithelium constituted less than 1% of the total basal cell population. SP cells constituted 0.5-3% of the total epithelial fraction. The SP transcriptome was essentially the same as ABCG2+ and both populations expressed genes indicative of a stem cell phenotype, however, the cells also expressed many genes in common with endothelial cells.
These results provide gene expression profiles for the prostate SP and ABCG2+ cells that will be critical for studying normal development and carcinogenesis, in particular as related to the cancer stem cell concept.
前列腺干细胞的鉴定与特性分析对于理解前列腺正常发育及致癌过程至关重要。基于流式细胞术的侧群(SP)技术已被用于在包括前列腺在内的多种人类组织类型中分离假定的成体干细胞。这种表型主要由ATP结合盒膜转运蛋白ABCG2介导。
对前列腺根治性切除术中获取的正常前列腺组织进行ABCG2免疫定位。分离正常人前列腺SP细胞和ABCG2阳性细胞,并用DNA阵列分析和逆转录聚合酶链反应测定基因表达。通过用CD31预分选去除内皮细胞。
ABCG2阳性细胞定位于前列腺基底上皮和内皮。基底上皮中的ABCG2阳性细胞占基底细胞总数的比例不到1%。SP细胞占上皮细胞总数的0.5% - 3%。SP转录组与ABCG2阳性细胞的基本相同,且这两个群体均表达指示干细胞表型的基因,然而,这些细胞也表达许多与内皮细胞共有的基因。
这些结果提供了前列腺SP细胞和ABCG2阳性细胞的基因表达谱,这对于研究正常发育和致癌过程,特别是与癌症干细胞概念相关的研究至关重要。