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目的探讨CD13、CD133双荧光标志流式细胞术(fluorescence activated cell sorter,FACS)分选人肝癌HuH7细胞系CD13+CD133+HCC细胞方法及癌干细胞(cancer stem cells,CSCs)特征。方法 CD13、CD133双标志,采用FACS从人肝癌HuH7细胞系分选出CD13+CD133+HCC、CD13+CD133-HCC、CD13-CD133+HCC、CD13-CD133-HCC等4种细胞亚群;通过CD13+CD133+HCC细胞生长曲线,细胞周期,形成肿瘤球和裸鼠体内成瘤能力,及对5-FU、吡柔比星化疗药物的敏感性研究,分析CD13+CD133+HCC细胞是否具有CSCs生物学特征。结果人肝癌HuH7细胞系CD13+CD133+HCC细胞占23.8%,3.1%分选为CD13+CD133+HCC细胞,78.45%的CD13+CD133+HCC细胞处于G0/G1期;CD13+CD133+HCC细胞增殖明显快于其他3个细胞亚群;在裸鼠103个CD13+CD133+HCC细胞就可以成瘤,而1.0×105个CD13-CD133-HCC只有2只成瘤(2/5),干细胞培养CD13+CD133+HCC细胞8～15 d形成肿瘤球;CD13+CD133+HCC细胞对5-FU和吡柔比星具有抵抗特性,而其他3个亚群较易于杀灭。结论 CD13、CD133双标志FACS从HuH7分选的CD13+CD133+HCC细胞具有CSCs特征,可能成为HCC治疗靶细胞。
Objective To investigate the method of sorting CD13 + CD133 + HCC cells and the characteristics of cancer stem cells (CSCs) by Huh7 cell line by fluorescence activated cell sorter (FACS) with CD13 and CD133. Methods CD13 + CD133 + HCC, CD13 + CD133-HCC, CD13-CD133 + HCC and CD13-CD133-HCC cell lines were isolated from human hepatocarcinoma HuH7 cell line by FACS. + CD133 + HCC cell growth curve, cell cycle, the formation of tumor spheres and tumorigenicity in nude mice, and the sensitivity to 5-FU and pirarubicin chemotherapeutic drugs, whether CD13 + CD133 + HCC cells have CSCs Learn characteristics. Results The percentages of CD13 + CD133 + HCC cells were 23.8% in HuH7 cell line, 3.1% in CD13 + CD133 + HCC cells, 78.45% in CD13 + CD133 + HCC cells in G0 / G1 phase. Significantly faster than the other three cell subsets; in 103 nude mice CD13 + CD133 + HCC cells can be tumor, and 1.0 × 105 CD13-CD133-HCC only 2 tumor (2/5), stem cells cultured CD13 + CD133 + HCC cells formed tumor spheres from 8 to 15 days. CD13 + CD133 + HCC cells exhibited resistance to 5-FU and pirarubicin, while the other three subpopulations were easier to kill. Conclusion CD13 and CD133 double markers FACS selected from HuH7 CD13 + CD133 + HCC cells have the characteristics of CSCs, may be the target of HCC treatment.