Laboratory of Oncology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, 1# Tiantan Xili, Beijing 100050, China.
Int J Biol Sci. 2012;8(7):992-1004. doi: 10.7150/ijbs.4454. Epub 2012 Aug 1.
EpCAM or CD133 has been used as the tumor initiating cells (TICs) marker in hepatocellular carcinoma (HCC). We investigated whether cells expressing with both EpCAM and CD133 surface marker were more representative for TICs in hepatocellular carcinoma Huh7 cells.
Four different phenotypes of CD133(+)EpCAM(+), CD133(+)EpCAM(-), CD133(-)EpCAM(+) and CD133(-)EpCAM(-) in Huh7 cells were sorted by flow cytometry. Then cell differentiation, self-renewal, drug-resistance, spheroid formation and the levels of stem cell-related genes were detected to compare the characteristics of TICs. The ability of tumorigenicity was measured in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice to verify TICs.
CD133(+)EpCAM(+) cells have many characteristics of TICs in Huh7 cells compared with CD133(+)EpCAM(-), CD133(-)EpCAM(+), CD133(-)EpCAM(-) cells, including enrichment in side population cells, higher differentiation capacity, increased colony-formation ability, preferential expression of stem cell-related genes, appearance of drug-resistant to some chemotherapeutics, more spheroid formation of culture cells and stronger tumorigenicity in NOD/SCID mice.
CD133(+)EpCAM(+) phenotype is precisely represented TICs in Huh7 cells. It might be useful for studying biology mechanism of TICs in hepatocellular carcinoma and screening new targets for cancer therapy.
EpCAM 或 CD133 已被用作肝癌(HCC)的肿瘤起始细胞(TICs)标志物。我们研究了在肝癌 Huh7 细胞中,同时表达 EpCAM 和 CD133 表面标志物的细胞是否更能代表 TICs。
通过流式细胞术对 Huh7 细胞中的四种不同表型的 CD133(+)EpCAM(+)、CD133(+)EpCAM(-)、CD133(-)EpCAM(+)和 CD133(-)EpCAM(-)进行分选。然后检测细胞分化、自我更新、耐药性、球体形成和干细胞相关基因的水平,以比较 TICs 的特征。在非肥胖糖尿病/严重联合免疫缺陷(NOD/SCID)小鼠中测量肿瘤形成能力,以验证 TICs。
与 CD133(+)EpCAM(-)、CD133(-)EpCAM(+)和 CD133(-)EpCAM(-)细胞相比,CD133(+)EpCAM(+)细胞在 Huh7 细胞中具有许多 TICs 的特征,包括富集于侧群细胞、更高的分化能力、增加的集落形成能力、优先表达干细胞相关基因、对某些化疗药物的耐药性、培养细胞的球体形成能力增强以及在 NOD/SCID 小鼠中的更强的致瘤性。
CD133(+)EpCAM(+)表型精确地代表了 Huh7 细胞中的 TICs。它可能有助于研究肝癌 TICs 的生物学机制,并筛选癌症治疗的新靶点。