干扰素α-2b联合5-氟尿嘧啶对人肝癌细胞生物学行为的影响

Effect of interferon α-2b combined with 5-fluorouracil on the biological behavior of hepatoma cells

目的检测干扰素(IFNα-2b)联合5-氟尿嘧啶(5-FU)在体外对肝癌细胞株HepG2的杀伤抑制作用,并探讨可能的作用机制。方法体外复苏、培养人肝癌细胞,并单用IFNα-2b或5-FU,以及不同浓度的IFNα-2b联合5-FU对其进行干预。用MTT法测定IFNα-2b联合5-FU对体外培养的人肝癌细胞生长的抑制率;用HOECHST染色法检测药物对人肝癌细胞凋亡的影响;用免疫细胞化学法检测用药后人肝癌细胞中突变型p53、c-myc表达的变化。结果 IFNα-2b和5-FU都能抑制HepG2细胞生长,并随着作用时间的延长而增强,IFNα-2b与5-FU联合用药比单用IFNα-2b或5-FU抑制效果更明显,并随着浓度的增加和作用时间的延长而增强;IFNα-2b和5-FU均能诱导肝癌细胞凋亡,二药联用时细胞凋亡更明显,可呈剂量依赖性地诱导肝癌细胞凋亡,不同浓度下的凋亡率均明显高于对照组;IFNα-2b和5-FU均可明显抑制突变型p53、c-myc蛋白的表达,二药联用抑制作用更明显。结论IFNα-2b和5-FU联用可呈剂量和时间依赖性地抑制肝癌细胞的增殖,呈剂量依赖性地诱导肝癌细胞凋亡,下调突变型p53、c-myc蛋白的表达,表现出协同作用。这些可能是IFNα-2b和5-FU联合抗癌的部分机制。

Objective To investigate the vitro inhibitory effect and mechanism of IFNα-2b combined with 5-fluorouracil on the hepatoma cell line HepG2.Methods Human liver cancer cells were recovered and cultured,and then IFNα-2b or 5FU alone and in different concentrations of IFNα-2b combined with 5-FU were used for intervention.Inhibition ratios of growth of liver cancer cells which were cultured in vitro were detected by MTT.The effect of apoptosis of IFNα-2b and 5FU on HepG2 cells was detected by HOECHST staining.The effect of the mutant p53 and c-myc proteins expression of IFNα-2b and 5-FU on cells were observed by immunocytochemistry and image analysis.Results IFNα-2b or 5-FU inhibited the growth of the human liver cancer cell line HepG2 in a time dependent manner.Compared with IFNα-2b or 5-FU alone,IFNα-2b combined with 5-FU significantly inhibited the growth of HepG2 cells in a dose-and-time dependent manner.The combination of IFNα-2b and 5-FU significantly increased the apoptosis ratio in a dose-dependent manner,and the apoptosis rates were significantly higher than that of control group.The combination of IFNα-2b and 5-FU significantly down-regulated the expression of mutant p53 and c-myc proteins compared with IFNα-2b or 5-FU alone.Conclusions IFNα-2b combined with 5-FU can significantly inhibit the growth of the human liver cancer cell line HepG2 in a dose-andtime dependent manner,induce apoptosis of hepatoma cells in a dose dependent manner and down-regulate expression of mutant p53 and c-myc proteins with a synergetic effect.These may be part of the anti-cancer mechanism of IFNα-2b combined with 5-FU.