Dou Jun, Wen Ping, Hu Weihua, Li Yating, Wu Yun, Liu Chunsheng, Zhao Fengshu, Hu Kai, Wang Jing, Jiang Chuilian, He Xiangfeng, Gu Ning
Department of Pathogenic Biology and Immunology, School of Basic Medical Science, Southeast University, 87# Dingjiaqiao Rd., Nanjing 210009, Jiangsu, China.
Cell Biol Int. 2009 Aug;33(8):807-15. doi: 10.1016/j.cellbi.2009.05.003. Epub 2009 May 27.
Increasingly more evidence shows that TSCs possess the characteristics of stem-like cells. However, a link between stem cells and TSCs remains to be shown. We have sorted SP cells and non-SP cells from the B16F10 cell lines by FACS, and then studied their cellular biological characteristics by using a SFCM culture method, proliferative assay in vitro, clone formation assays in soft agar and normal media, tumorigenic assays in C57BL/6 mice, and resistance to chemotherapy assay in vitro, the quantitative detecting expression of ABCG2 and their CD phenotype analysis by a FCM. We detected 0.96% SP cells in the B16F10 cells and found that they had obvious differences in characteristics from non-SP cells. They possessed a marked capacity for self-renewal in soft agar and culture medium, strong tumorigenic potential in C57BL/6 mice, apparent resistance to vinblastin in vitro, upregulated ABCG2 expression, and a high expression of CD44+CD133+CD24+ phenotypes. We conclude that there were a few of SP cells that had the characteristics of tumor stem-like cells which may provide a useful tool and a readily accessible source for further study when specific TSCs markers are unknown.
越来越多的证据表明，肿瘤干细胞具有干细胞样细胞的特征。然而，干细胞与肿瘤干细胞之间的联系仍有待证实。我们通过荧光激活细胞分选术（FACS）从B16F10细胞系中分选了侧群（SP）细胞和非SP细胞，然后使用无血清细胞培养基（SFCM）培养方法、体外增殖试验、软琼脂和正常培养基中的克隆形成试验、C57BL/6小鼠体内致瘤试验以及体外化疗耐药试验，并通过流式细胞术（FCM）定量检测ABCG2的表达及其CD表型分析，研究了它们的细胞生物学特性。我们在B16F10细胞中检测到0.96%的SP细胞，发现它们与非SP细胞在特性上有明显差异。它们在软琼脂和培养基中具有显著的自我更新能力，在C57BL/6小鼠中具有很强的致瘤潜力，在体外对长春花碱有明显抗性，并上调了ABCG2表达，且高表达CD44+CD133+CD24+表型。我们得出结论，有少数SP细胞具有肿瘤干细胞样细胞的特征，在特定肿瘤干细胞标志物未知时，这可能为进一步研究提供有用的工具和易于获取的来源。
