5-氟尿嘧啶作用人肝癌细胞系SMMC-7721后残存细胞中肿瘤干细胞比例增加

目的主要探讨5-氟尿嘧啶(5-FU)作用肝癌细胞系SMMC-7721后残余细胞中NCAM/CD56、ICAM-1/CD54、EpCAM、CD133及ABCG2阳性细胞表达率的变化。方法检测不同浓度5-FU作用于SMMC-7721细胞系前后细胞增殖能力的变化,流式细胞仪检测残存细胞的细胞周期分布以及5种表面标志物阳性细胞表达率的变化。结果不同浓度5-FU均可抑制SMMC-7721增殖。使用10μg/mL的5-FU浓度作用48 h进一步研究,SMMC-7721细胞周期各期细胞比例均较对照组有影响,且G0/G1、S和G2/M期5-FU作用前后差异有统计学意义(P<0.05),G0/G1期阻滞明显。5-FU处理使SMMC-7721 5种细胞表面标志物阳性细胞比例均明显升高,差异均具有统计学意义(P<0.05),阳性细胞比例均明显升高。结论上述实验提示肝癌干细胞逃避5-FU单药物的杀伤可能是临床上肝癌化疗失败和肿瘤复发的重要原因,同时,此实验方法可以作为肝癌干细胞富集的科研模型。

The proportion of tumor stem cells increases in the presence of 5-FU on human liver carcinoma SMMC-7721 cell line

Objective To study the changes of NCAM/CD56,ICAM-1/CD54,EpCAM,CD133 and ABCG2 positive cells proportion in the presence of 5-FU on human liver carcinoma SMMC-7721 cell line. Methods Cell proliferation in different concentrations of 5-FU was evaluated by CCK-8(cell counting kit-8).Cell cycle and positive cells proportion of NCAM,ICAM-1,EpCAM,CD133,ABCG2 were measured by Flow Cytometry in an appropriate concentration of 5-FU. Results The proliferation of liver carcinoma cell was inhibited by different concentrations of 5-FU.The appropriate concentration of 5-FU was determined as 10 μg/mL with 48 h incubation.After treatment,G0/G1 phase cells increased from 46.617%±18.857% to 83.917%±12.996%.Whereas,S phase cells decreased from 50.510%±16.405% to 15.360%±13.517%;G2/M phase cells decreased from 2.872%±2.824% to 0.722%±0.711%.The proportion of G0/G1,S and G2/M stage compared with control group changed with statistic differences(P<0.05).Meanwhile,positive cells proportion of cell surface makers NCAM,ICAM-1,EpCAM,CD133,ABCG2 increased from 0.6%±0;78.0%±0.9%;3.9%±0.1%;8.2%±0.6%;0.3%±0.1% to 7.2%±0.1%;94.6%±0.1%;5.4%±0.3%;17.5%±0.4%;1.8%±0.1%,respectively.The difference was significant(P<0.05).NCAM,ICAM-1,EpCAM,CD133 and ABCG2 positive cells proportion increased in the presence of 5-FU on human liver carcinoma SMMC-7721 cell line. Conclusion It suggests that the escape mechanism of liver cancer stem cells in the presence of 5-FU is the major reason of chemotherapy failure clinically.Meanwhile,this experimental method could be a researchful model in enriching liver cancer stem cells.