肝癌细胞系Huh-7中CD133阳性细胞亚群的分离及其特性的研究

Isolation of CD133 positive cells and stem cell property from hepatocarcinoma Huh-7 cell line

目的:研究CD133在人肝癌细胞系Huh-7中的表达,初步探讨肝癌中CD133阳性细胞亚群的干细胞特性。方法:流式细胞荧光激活分选技术纯化肝癌细胞系Huh-7中CD133肿瘤细胞,体外培养并观察其增殖、体内成瘤及分化能力。结果:流式细胞仪检测肝癌细胞系Huh-7中有62.3%的细胞CD133呈阳性表达,流式细胞荧光激活分选的CD133肿瘤细胞在无血清培养基中第3、5、7d的吸光度(OD值)分别为0.310、0.362、0.564,均高于相同条件下未分选细胞和CD133阴性细胞;CD133阳性细胞亚群成瘤能力明显强于阴性细胞亚群;CD133阳性细胞亚群在培养体系中的比例逐日下降,至培养的第8天,由第1天的92.3%下降至61.4%。结论:肝癌细胞系Huh-7中CD133阳性细胞亚群比其它细胞亚群具有较强增殖、成瘤和分化能力,是具有肝癌干细胞特性的细胞亚群。

Objective：To detect the expression of CD133 in hepatocellular carcinoma Huh -7 cell line and study its stem cell property. Methods：Fluorescence -activated cell sorting（FACS） was used to detect the expression of putative tumor- initiating cell marker CD133 in Huh -7 cell line. CD133 ^＋ and CD133^- tumor ceils were cultured and their ability of proliferation and differentiation were observed in vitro, tumor cell implantation experiments were done to test the tumorigenicity of the above two suhpopulations. Results：CD133 was detected in 62.3% of Huh -7 cell line. In serum-free RPMI 1640, on day 3,5 and 7, their UV absorptions were 0. 310,0.362 and 0.564 respectively, compared with CD133^- cells and control Huh -7 cells, CD133^＋ cells demonstrated increased proliferation capacity. Tumor cell implantation experiments showed that only 1 × 10^3 CD 133 ^＋ cells were sufficient for tumor formation （2/10） ,whereas an injection of 1 × 10^5 CD133^ - cells could initiate tumors （2/10）. The proportion of CD133^＋ cells decreased in culture as days passed by, in eight days of culture, the percentage of CD 133 ^＋ cells decreased from 92.3 % to 61.4%. Condusion：CD133 ^＋ cell subpopulation has stronger ability to proliferate, differentiate in vitro and to form tumors in vivo than other subpopulations in Huh -7 cell line. In one word, CD133^ ＋ cell subpepulation has stem cell property.