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目的探讨干扰素(IFN)-α联合5-氟尿嘧啶(FU)对人肝癌HepG-2细胞株DLC-1和Bcl-2表达的影响。方法培养人肝癌细胞HepG-2,将其分为对照组,5-FU组,IFN-α组及联合用药组,采用噻唑蓝(MTT)实验法观察细胞增殖,流式细胞仪(FCM)检测细胞周期和细胞凋亡,反转录-聚合酶链反应(RT-PCR)检测肝癌缺失基因DLC-1、凋亡抑制基因Bcl-2表达变化。结果IFN-α单独应用对HepG-2细胞增殖抑制作用不明显,与5-FU联合应用时抑制作用明显增强,经药物处理48h后,联合用药组的细胞凋亡率为(16.27)%,与对照组及IFN-α和5-FU单独应用组相比增高。联合用药组HepG-2细胞DLC-1的表达较对照组和单药组明显上升,Bcl-2的表达较对照组和单药组明显下降。结论IFN-α和5-FU联合应用可明显抑制HepG-2细胞的增殖并诱导凋亡,其机制之一可能是调节肝癌缺失基因DLC-1的表达,引起了天门冬氨酸(caspase-3)介导的细胞凋亡;调节凋亡抑制基因Bcl-2的表达,影响内源性凋亡信号传导通路来发挥作用,并且两者之间存在一定的协同效应。
Objective To investigate the effect of interferon (IFN) -α combined with 5-fluorouracil (FU) on the expression of DLC-1 and Bcl-2 in human hepatoma HepG-2 cell line. Methods HepG-2 cells were cultured and divided into control group, 5-FU group, IFN-α group and combination group. Cell proliferation was detected by MTT assay. Flow cytometry (FCM) Cell cycle and apoptosis were detected by flow cytometry. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of DLC-1 and Bcl-2. Results The inhibitory effect of IFN-α alone on HepG-2 cell proliferation was not obvious. When combined with 5-FU, the inhibitory effect of IFN-α alone was significantly enhanced. After 48 h of treatment, the apoptosis rate of combined treatment group was (16.27)%, The control group and IFN-α and 5-FU alone application group increased. The expression of DLC-1 in HepG-2 cells in combination group was significantly higher than that in control and monotherapy groups, and the expression of Bcl-2 was significantly lower than that in control and monotherapy groups. Conclusion The combination of IFN-α and 5-FU can significantly inhibit the proliferation and induce the apoptosis of HepG-2 cells. One of the mechanisms may be the regulation of the expression of DLC-1, a gene lacking in HCC, and caspase-3 ) -mediated apoptosis, regulate the expression of apoptosis-inhibiting gene Bcl-2 and influence the signal transduction pathway of endogenous apoptosis, and there is a certain synergistic effect between the two.