EpCAM和CD13在不同肝癌细胞系中的表达和乙型肝炎病毒X蛋白对其表达的影响

The Expression of Liver Cancer Stem Cell Marker EpCAM and CD13 in Different Liver Cancer Cell Lines and the Influence of Hepatitis B Virus X Protein on Their Expression

目的:探讨肝癌干细胞标记物EpCAM和CD13在不同肝癌细胞系中的表达及乙型肝炎病毒X蛋白(HBx)对EpCAM和CD13表达的影响。方法:Western blot检测正常肝细胞系(LO2细胞)和肝癌细胞系(Huh7,Bel-7404,Hep3B,QSG-7701细胞)中肝癌干细胞标记物EpCAM和CD13的表达情况,同时建立稳定转染HBx基因的Bel-7404(Bel-7404/HBx)细胞系,用Real-time PCR和Western blot检测转染后EpCAM和CD13表达情况。结果:EpCAM和CD13在不同的肝癌细胞系中表达水平完全不同;转染HBx基因后EpCAM的mRNA和蛋白表达水平明显上调,分别为对照组的(2.04±0.16)和(2.03±0.25)倍;而CD13的mRNA和蛋白表达水平明显下调,分别为对照组的(69±8)%和(55±7)%。结论:肝癌干细胞可能存在多个干细胞亚群,在不同的肝癌细胞中干细胞标记物的表达水平不同,而HBX对不同亚群的影响也不同。

Objective： To investigate the expression of liver cancer stem cell marker EpCAM and CD13 in different liver cancer cell lines and the influence of Hepatitis B virus X protein on their ex- pression. Methods： Western blot was adopted to investigate the expression of liver cancer stem cell markers EpCAM and CD13 in normal liver cell line （LO2） and liver cancer cell lines （Huh7, Bel- 7404, Hep3B, QSG-7701 ）. Meanwhile a stable transfection with HBx gene cell line BEL-7404/HBX was established. The expression of EpCAM and CD13 were detected by Real-time PCR and Western blot after transfection. Results： The tendency of EpCAM and CD13 expression are completely different in different liver cancer cell lines. After HBx gene transfection, the expression of EpCAM mRNA level and protein level were both increased, by 2.04 ± 0. 16 and 2.03 ± 0.25 to the control group （ P 〈 0. 05 ） ; while the expression of CD13 level and protein level were both decreased, by （69 ± 8）% and （55 ± 7 ） % to the control group （ P 〈 0.05 ）. Conclusions： There may be multiple subgroups of liver stem ceils in liver cancer stem cell, and influence of Hepatitis B virus X protein on them is varied.