摘要
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 explore the effect of IFN-α combined with 5-Fluorouracil （5-FU） on expression of DLC-1 and Bcl-2 gene of hepataocellular carcinoma cell line HepG-2. Methods HepG-2 cells were divided into 4 groups, namely the control group, IFN-α group, 5-FU group and combined treatment group. Cell proliferation was e- valuated by MTT assay. Cell cycle and apoptosis ratio were observed by flow cytometry. Expression of mRNA DLC-1 and Bcl-2 gene was measured by RT-PCR. Results The inhibition effect of 5-FU plus IFN-α was significantly stronger than INF-α and 5-FU alone. After treatment for 48 h, the rate of apoptosis was 16.27% in the combined treatment group, significantly higher than as found for the control group, IFN-α group and 5-FU group. The expression of DLC-1 was higher and the expression of Bcl-2 was lower in the combined treatment group than those in the control group, IFN-α group or 5-FU group. Conclusion Combination of IFN-α and 5-FU may evidently suppress the proliferation and induce apoptosis of HepG-2 cells. The mechanisms may be up-regulation of DLC-1 whieh pro- vokes aspartate （caspase-3）-mediated apoptosis and down-regulation of Bcl-2 gene which affects endogenous signal pathway of apoptosis, with some synergistic effect between the both.
出处
《中国药物与临床》
CAS
2010年第1期28-31,共4页
Chinese Remedies & Clinics
基金
山西省教育厅科研基金(20091172)
