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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。
Objective To investigate the changes of NCAM / CD56, ICAM-1 / CD54, EpCAM, CD133 and ABCG2 positive cells in 5-fluorouracil (5-FU) -treated hepatoma cell line SMMC-7721. Methods The proliferation of SMMC-7721 cells was detected by different concentrations of 5-FU. The cell cycle distribution of remaining cells and the expression of 5 kinds of surface markers were detected by flow cytometry. Results Different concentrations of 5-FU could inhibit the proliferation of SMMC-7721. After 5-FU concentration of 10μg / mL for 48h, the percentage of cells in SMMC-7721 cell cycle was significantly higher than that in control group, and the difference was statistically significant before and after 5-FU in G0 / G1, S and G2 / M Significance (P <0.05), G0 / G1 phase block was obvious. 5-FU treatment significantly increased the percentage of 5 cell surface marker positive cells in SMMC-7721 cells, the difference was statistically significant (P <0.05), and the percentage of positive cells was significantly increased. Conclusions The above experiments suggest that the killing of hepatocellular carcinoma stem cells by 5-FU may be the main reason for the failure of chemotherapy and recurrence of hepatocellular carcinoma in clinic. At the same time, this method can be used as a research model for the enrichment of hepatocellular carcinoma stem cells.