芬太尼和瑞芬太尼对人肝癌细胞SMMC-7721生物学特性的影响

Effects of fentanyl and remifentanil on biological characteristics of human hepatoma cell line SMMC-7721

目的观察芬太尼和瑞芬太尼对人肝癌细胞SMMC-7721增殖、凋亡以及迁移能力的影响。方法体外培养人肝癌细胞SMMC-7721至对数期,接种到96孔板或6孔板中,随机均分为11组:1个空白对照组(N组)、5个芬太尼组(F1、F2、F3、F4、F5组)和5个瑞芬太尼组(R1、R2、R3、R4、R5组)。F1、F2、F3、F4、F5组对应的芬太尼终浓度为0.5、5、50、500、1 000ng/ml,R1、R2、R3、R4、R5组对应的瑞芬太尼终浓度为0.5、5、50、500、1 000ng/ml,每组设6个平行副孔,N组加入等剂量的培养基。每组均孵育24、48和72h,用CCK-8法检测细胞的增殖活性;孵育24h时应用流式细胞仪检测细胞凋亡情况和细胞周期分布;孵育48h时应用划痕实验检测细胞迁移能力。结果与N组比较,当药物浓度≥5ng/ml时随着浓度的增高芬太尼组和瑞芬太尼组的细胞活性逐渐降低,细胞凋亡率逐渐升高,G0/G1期比例逐渐增加,S期比例逐渐减少,细胞迁移能力逐渐降低(P<0.05)。结论芬太尼和瑞芬太尼能剂量依赖性地抑制人肝癌细胞SMMC-7721的增殖,降低该肝癌细胞的迁移能力,可能与促进细胞凋亡,影响细胞周期有关。

Objective To investigate the effect of different doses of fentanyl and remifentanil on human hepatoma cell line SMMC-7721 proliferation,apoptosis,invasiveness and migration capability.Methods SMMC-7721 cells cultured to logarithmic phase were seeded in 96-well or 6-well culture plate and randomly divided into 11 groups.The experimental group including：blank control group （N）,5 groups of fentanyl （F） and 5 groups of fentanyl （R）.Different concentrations of fentanyl were 0.5,5,50,500,1 000 ng/ml （groups F1,F2,F3,F4,F5） and remifentanil were 0.5,5,50,500,1 000 ng/ml（groups R1,R2,R3,R4,R5）.Each group set up six parallel wells.Added isopyknic cell culture fluid to the blank control group wells.CCK-8 method to detect cell proliferation activity at incubated 24,48,72 h respectively; flow cytometry instrument to detect cell apoptosis and cell cycle at incubated 24 h; the scratches experimental determination of cell migration ability at incubated 24 h.Results Compared with group N,fentanyl group and remifentanil group cell activity decreased,cell migration ability decreased,apoptosis rate increased,G0/G1 phase increased and S phase decreased with the concentration increased significantly （P＜0.05）.Conclusion Fentanyl and remifentanil can inhibit human hepatoma cell SMMC 7721 proliferation,reduce cell migration ability,may be related to promote cell apoptosis and impact cell cycle.