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目的探讨重组干扰素-α(IFN-α)对肿瘤抑制作用的靶分子及可能作用机制。方法选用HepG2肝癌细胞系,分别应用1000 U/ml、2000 U/ml浓度的IFN-α作用于HepG2细胞,在药物作用6 h、16 h后应用RT-PCR;Dickkopf-1(DKK-1)方法分别检测各实验组DKK-1 mRNA表达水平,Western blot杂交法检测DKK-1蛋白表达水平。结果定量RT-PCR结果显示,IFN-α处理能增强DKK-1mRNA表达。2000 U/mL IFN-α作用HepG2细胞16 h组其DKK-1 mRNA表达(0.00039±0.000057)高于处理6 h组(0.000195±0.000021,t=6.994,P=0.0022)以及1000 U/mL作用16 h组(0.000307±0.000012,t=2.876,P=0.0452);Western blot显示重组IFN-α作用于HepG2细胞后,均能够导致细胞内DKK-1蛋白表达增强(P值均<0.05)。结论 IFN-α能够上调DKK-1表达,DKK-1可能是IFN-α作用的下游调节靶基因,这可能是IFN-α发挥抗肿瘤作用机制之一。
Objective To investigate the target of recombinant interferon-α (IFN-α) on tumor inhibition and its possible mechanism. Methods HepG2 hepatocarcinoma cell lines were selected and HepG2 cells were treated with 1000 U / ml and 2000 U / ml of IFN-α respectively. RT-PCR and Dickkopf-1 (DKK- Methods The DKK-1 mRNA expression levels in each experimental group were detected, and the protein expression of DKK-1 was detected by Western blot. Results The results of quantitative RT-PCR showed that IFN-α treatment enhanced DKK-1 mRNA expression. The expression of DKK-1 mRNA (0.00039 ± 0.000057) in HepG2 cells treated with 2000 U / mL IFN-α for 16 h was higher than that treated with 6 h (0.000195 ± 0.000021, t = 6.994, P = 0.0022) h group (0.000307 ± 0.000012, t = 2.876, P = 0.0452). Western blot showed that the expression of DKK-1 protein in HepG2 cells was enhanced by recombinant IFN-α (all P <0.05). Conclusions IFN-α can up-regulate the expression of DKK-1, and DKK-1 may be a downstream regulatory target of IFN-α. This may be one of the mechanisms by which IFN-α exerts antitumor effects.