Characterization of the Hoechst 33342 side population from normal and malignant human renal epithelial cells
Characterization of the Hoechst 33342 side population from normal and malignant human renal epithelial cells
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DOI：
10.1152/ajprenal.90286.2008
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发表时间：
2008-09-01
影响因子：
4.2
通讯作者：
Clarke, Noel W.
中科院分区：
文献类型：
--
作者：
Addla, Sanjai K.;Brown, Mick D.;Clarke, Noel W.
肾细胞癌（RCC）的基本变化是不好的特点。据推测，“癌症干细胞”可能在致癌作用中有影响，并且在其他上皮癌中，上皮侧群（SP）富含干细胞样细胞。在这项研究中，我们已经分离和特点的SP和非SP（NSP）人口正常（NK）和恶性（RCC）的人肾组织。NK样本取自接受非肾癌手术的患者，配对的恶性和肉眼可见的正常组织样本取自接受RCC手术的患者。Hoechst 33342染料流出技术用于从正常和恶性人肾组织中分离上皮SP和NSP。细胞亚群的谱系，细胞周期，和假定的干细胞标志物的表型，并在体外集落形成和增殖试验的功能特征。在NK和RCC中，SP分别占上皮细胞的3.8 +/- 0.4和5.9 +/- 0.9%，其中14.1 +/- 3.5和13.2 +/- 3.6%显示为G（0）。SP细胞在集落形成效率、长期培养和球状体测定中表现出更大的增殖潜力，并在组织培养传代后得以维持。我们已经表明，肾SP是丰富的静止细胞，具有高增殖能力和干细胞特性。然而，该群体是异质的，证实了术语“SP细胞”和“干细胞”不能互换使用。
The fundamental changes which predispose for renal cell carcinoma (RCC) are poorly characterized. It is hypothesized that "cancer stem cells" may be influential in carcinogenesis, and the epithelial side population (SP) is enriched for stemlike cells in other epithelial cancers. In this study, we have isolated and characterized the SP and non-SP (NSP) populations from normal (NK) and malignant (RCC) human kidney tissue. NK specimens were taken from patients undergoing non-renal cancer surgery and paired malignant and macroscopically normal tissue samples were taken from patients undergoing surgery for RCC. The Hoechst 33342 dye efflux technique was used to isolate epithelial SP and NSP from normal and malignant human renal tissue. Cellular subpopulations were phenotyped for lineage, cell cycle, and putative stem cell markers, and functionally characterized using in vitro colony-forming and proliferation assays. The SP constituted 3.8 +/- 0.4 and 5.9 +/- 0.9% of epithelial cells in NK and RCC, respectively, of which 14.1 +/- 3.5 and 13.2 +/- 3.6% were shown to be in G(0). SP cells demonstrated greater proliferative potential in colony-forming efficiency, long-term culture, and spheroids assays and were shown to be maintained upon tissue culture passage. We have shown that the renal SP is enriched for quiescent cells, with a high proliferative capacity and stemlike properties. The population is, however, heterogeneous, confirming that the terms "SP cell" and "stem cell" cannot be used interchangeably.