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目的:探讨α-干扰素(IFN-α)联合5-FU对肝癌细胞HepG2·2·15生物学特性的影响及其作用机制。方法:采用MTT法观察5-FU和IFN-α对HepG2·2·15细胞增殖的影响,流式细胞术检测细胞周期和细胞凋亡变化及细胞表面Fas表达变化;RT-PCR检测凋亡抑制基因Bcl-xL表达变化。结果:IFN-α单独应用对HepG2·2·15细胞增殖抑制作用较弱,与5-FU联合应用时对细胞的增殖抑制作用明显增强,P<0·01;两者联合应用对HepG2·2·15细胞周期的影响表现为与对照组和IFN-α、5-FU单用药组相比,联合用药组G0/G1期细胞比率升高(78·33%vs54·95%、60·08%和72·50%),S期比率降低(10·97%vs29·04%、18·69%和17·44%),P<0·01;经药物处理48h后,联合用药组的细胞凋亡率为(25·08±2·54)%,与对照组及IFN-α和5-FU单独应用组相比,细胞凋亡率明显增高,P<0·01。IFN-α对HepG2·2·15细胞表面Fas表达没有影响,但可协同5-FU明显提高细胞表面Fas的表达,P<0.01。联合用药组HepG2·2·15细胞Bcl-xL的表达较对照组和单药组明显下降,P<0·05。结论:IFN-α和5-FU联合应用可抑制HepG2·2·15细胞的增殖并诱导凋亡,其机制之一可能是调节凋亡抑制基因Bcl-xL的表达,影响内源性凋亡信号传导通路来发挥作用。
Objective: To investigate the effects of interferon-α (IFN-α) and 5-FU on the biological characteristics of HepG2 · 2.15 hepatocellular carcinoma cells and its mechanism. Methods: MTT assay was used to observe the effects of 5-FU and IFN-α on the proliferation of HepG2.2.15 cells. Flow cytometry was used to detect the changes of cell cycle and apoptosis and the expression of Fas on the cell surface. Gene Bcl-xL expression changes. Results: The inhibition of HepG2.2.15 cell proliferation by IFN-α alone was weaker than that of HepG2-2.15 cells alone. The combination of 5-FU and IFN-α significantly inhibited the proliferation of HepG2-2.15 cells, P <0.01; · 15 cell cycle showed that the cell ratio of G0 / G1 phase increased (78.33% vs54.95%, 60.08%) in the combination group compared with the control group and the IFN-α and 5-FU monotherapy groups And 72.5% respectively), the S phase ratio decreased (10 · 97% vs 29.04%, 18.69% and 17.44%, P <0.01) The apoptosis rate was (25.08 ± 2.54)%. Compared with the control group and IFN-α and 5-FU alone groups, the apoptosis rate was significantly increased, P <0.01. IFN-α had no effect on the expression of Fas on the surface of HepG2-2.15 cells, but synergistic effect of 5-FU on cell surface Fas expression was significant (P <0.01). The expression of Bcl-xL in HepG2 · 2 · 15 cells in combination group was significantly lower than that in control group and single drug group (P <0.05). CONCLUSION: The combination of IFN-α and 5-FU can inhibit the proliferation and induce apoptosis of HepG2 · 2.15 cells. One of the mechanisms may be the regulation of the expression of Bcl-xL, which affects the expression of endogenous apoptosis signal Conduction path to play a role.