尼妥珠单抗联合表阿霉素对人肝癌细胞株HepG2体外生长的影响

The effect of epirubici combined with nimotuzumab on human hepatocellular carcinoma cell line HepG2 in vitro

目的观察尼妥珠单抗(h-R3)与表阿霉素(EPI)联合对人肝癌细胞株HepG2体外生长及凋亡的影响。方法选用浓度递增的h-R3和EPI,单独和联合作用于HepG2细胞,MTT法观察不同时间点、不同药物剂量对HepG2细胞生长的影响,计算两者联合的q值;流式细胞仪检测两种药物对HepG2细胞凋亡率及细胞周期分布情况的影响。结果 h-R3和EPI单药对HepG2细胞生长均有抑制作用且呈时间和剂量依赖性,两药单独作用72h后对HepG2细胞生长的最高抑制率分别为(49.56±8.93)%和(92.97±1.19)%,两药联合对HepG2细胞生长的最高抑制率为(96.44±1.0)%,联合用药可提高细胞增殖抑制率,呈相加作用;流式细胞仪检测发现,联合组细胞凋亡率高于单药组,随时间增加凋亡率呈升高趋势;细胞周期检测提示h-R3使细胞阻滞在G0/G1期,EPI阻滞细胞在S期,两药均能影响细胞周期分布。结论 h-R3在体外与EPI联合可以提高对HepG2细胞增殖抑制及凋亡的作用,两药均可影响HepG2细胞周期的分布。

Objective To investigate the anti-proliferation effect of epidermal growth factor receptor inhibitor nimotuzumab combined with chemotherapeutic drug epirubici on growth of human hepatocellular carcinoma HepG2 cells.Methods MTT assay was performed to evaluated the growth inhibitory rate of HepG2 cells.Increasing dose of nimotuzumab and epirubici alone or in combination were administrated to HepG2 cells.The inhibitory effects of the drugs on cell proliferation at different time points were observed,and q value was calculated when the two drugs combined.Different drug alone or combination effect on HepG2 cells apoptosis and cell cycle changes were determined by flow cytometry.Results Nimotuzumab and epirubici both inhibited the growth of HepG2 cells in a time-and dose-dependent manner.The single drug of nimotuzumab or epirubici had effect on HepG2 cells,after 72-hour treatment,the proliferation inhibition rate of HepG2 cells was（49.56±8.93）% and（92.97±1.19）%.Combination of nimotuzumab and epirubici could increase the proliferation inhibition rate of HepG2 cells,which was（96.44±1.0）% after 72-hour treatment.And the two drugs had synergistic role on the proliferation inhibition by q value.From flow cytometry result,after the drug nimotuzumab and epirubici alone or combined treatment,the apoptosis ratio of combination group was higher,and the apoptosis ratio was increased by treatment time prolonged.Nimotuzumab had effect on arresting the cells in G0/G1 phase while epirubici arrested the cells in S phase by cell cycle evaluated,which suggested the two drugs could both effect cell cycle changes of HepG2 cells.Conclusion Nimotuzumab combined with epirubici can enhance the therapeutic effects on HepG2 cells in vitro,which increase the sensitivity of HepG2 cells to epirubici.And both of the two drugs have effect on cell cycle changes of HepG2 cells.