HuH7细胞系CD13_CD133_HCC细胞分选及CSCs特性分析

HuH7细胞系CD13+CD133+HCC 细胞分选及CSCs 特性分析

金世龙，黄中荣，陈

华，余天雾，曹洪，龙运全，周健，李鹤，苟毅，廖娟（402160重庆，重庆医科大学附

属永川医院肝胆外科）

［摘要］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G 0/G 1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1.0ˑ105个

CD13－CD133－HCC 只有2只成瘤（2/5），干细胞培养CD13+CD133+HCC 细胞8 15d 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。

［关键词］肝癌；癌干细胞；CD13；CD133

［中图法分类号］R329-

33；R730．23；R735．7［文献标志码］A

［通信作者］黄中荣，E-mail ：hzr1966@126．com

Sorting and cancer stem cell characteristic analysis of CD13+CD133+HCC cells in HuH7cell line

Jin Shilong ，Huang Zhongrong ，Chen Hua ，Yu Tianwu ，Cao Hong ，Long Yunquan ，Zhou Jian ，Li He ，Gou Yi ，Liao Juan （Department of Hepatiobiliary Surgery ，Yongchuan Hospital ，Chongqing Medical University ，Chongqing ，402160，China ）

［Abstract ］Objective To study the sorting of CD13+CD133+HCC cell fraction in HuH7cell line by fluorescence activated cell sorter （FACS ）based on the expression of both CD13and CD133markers ，and to analyze the cancer stem cells （CSCs ）characteristics of CD13+CD133+HCC cells．Methods The antibodies against CD13and CD133were used as CSCs marker to sort CD13+CD133+HCC cells ，CD13+CD133－HCC cells ，CD13－CD133+HCC cells and CD13－CD133－HCC cells from HuH7cell line by FACS．The growth curve ，phase of cell cycle ，sphere formation ability in vitro ，tumor formation ability in vivo and susceptibility to 5-FU and pirarubicin of the CD13+CD133+HCC cells were investigated for CSCs identification and character-ization of the CD13+CD133+HCC cells．Results The CD13+CD133+HCC cells accounted for 23．8%in HuH7cell line ，and 3．1%of the CD13+CD133+HCC cells was sorted as CSC-like cells．There was 78．45%

of the CD13+CD133+HCC cells existed in G 1/G 0phase．The proliferating ability of the CD13+CD133+HCC cells was significantly stronger than that of the other cell subsets．The CD13+CD133+HCC cells could form a tumor in naked mice after 4weeks with a number of 103，but 1．0ˑ105of the CD13－CD133－HCC cells were needed to form a tumor．The in vitro sphere formation of the CD13+CD133+HCC cells was observed after 8to

15d．The CD13+CD133+HCC cells had the chemo-resistant ability to 5-FU and pirarubicin ，but the other cell subsets could be killed easily by these drugs．Conclusion The CD13+CD133+HCC cells sorted from HuH7

cell line by FACS based on the expression of both CD13and CD133markers have the characteristics of CSCs ，and may be the therapeutic target for human hepatocellular carcinoma．

［Key words ］liver cancer ；cancer stem cells ；CD13；CD133

Corresponding author ：Huang Zhongrong ，E-mail ：hzr1966@126．com