肝癌细胞系Huh-7中EpCAM^+细胞亚群的肿瘤干细胞样特性

Cancer stem cell-like properties of EpCAM^+ cell subpopulation in hepatocellular carcinoma cell line Huh-7

目的:观察从肝癌细胞系Huh-7中分选出的EpCAM+细胞亚群的肿瘤干细胞样特性。方法:采用免疫磁珠法从Huh-7细胞中分离EpCAM+和EpCAM-细胞亚群,比较其体外增殖、克隆形成、分化、耐药和体内成瘤能力。结果:EpCAM+细胞体外增殖能力强于EpCAM-细胞(P<0.001);EpCAM+细胞克隆形成数高于EpCAM-细胞(P<0.001);分选后随着培养时间的延长,EpCAM+细胞的比率逐渐降低;加入顺铂培养后,EpCAM+细胞亚群的比率增加(P<0.001);1×103个EpCAM+细胞接种裸鼠皮下4周即可成瘤,而在相同条件下,EpCAM-细胞至少需要接种1×105个和接种更长的时间才能成瘤。结论:EpCAM+细胞亚群具有肝癌干细胞样特性,EpCAM可作为分离纯化肝癌干细胞的一个标志物。

Aim：To observe the properties of epithelial cell adhesion molecule（EpCAM）-positive cell subpopulation sorted from human hepatocellular carcinoma cell line Huh-7.Methods：EpCAM＋ and EpCAM-subpopulations were isolated by automagnetic activated cell-sorting system and their abilities of proliferation,differentiation,colony formation,resistance to cisplatin and tumorgenicity were observed.Results：The cell proliferation ability of EpCAM＋ subpopulation was stronger than that of EpCAM-subpopulation（P0.001）.The number of colonies formed by EpCAM＋ subpopulations was more than that of EpCAM-subpopulations（P0.001）.The proportion of EpCAM＋ cells in sorted subpopulation decreased gradually as the cells were cultured.After Huh-7 cells were cultured with cisplatin,the percentage of EpCAM＋ cells increased（P0.001）.Only 103 EpCAM＋ cells were enough to form tumors in nude mice after subcutaneous inoculation for four weeks,while 105 EpCAM-cells were necessary to form tumors.Conclusion：EpCAM＋ subpopulation sorted from Huh-7 has properties of liver cancer stem cell.EpCAM might be a valuable marker for isolating and purifying hepatocellular carcinoma stem cells.