Myxovirus resistance protein A(MxA)induces the expression of interferon-stimulated genes in HepG2 cells by enhancing the activity of the interferon-stimulated response element(ISRE)
Objective To explore the effects of Myxovirus resistance protein A(MxA)on the Janus kinase/Signal transducer and activator of transcription(JAK/STAT)pathway in HepG2 cells.Methods HepG2 cells were transfected with the pcDNA3.1-Flag-MxA construct,and subsequent localization and expression of the MxA protein were detected through immunofluorescence cytochemistry.The presence of MxA protein was further confirmed by using Western blot analysis.Following transfection with MxA small interfering RNA(si-MxA)and subsequent treatment with alpha interferon(IFN-α),real-time fluorescent quantitative PCR was employed to measure the mRNA levels of myxovirus resistance protein A(MxA),protein kinase R(PKR),and oligoadenylate synthase(OAS).Western blot analysis was used to detect the protein expression of MxA,PKR,OAS,signal transducer and activator of transcription 1(STAT1),phosphorylated STAT1(pSTAT1),STAT2,phosphorylated STAT2(p-STAT2)and interferon regulatory factor 9(IRF9).Additionally,pcDNA3.1-Flag-MxA and plSRE-TA-luc were co-transfected into HepG2 and HepG2.2.15 cells,respectively,to assess the activity of the interferon-stimulated response element(ISRE)by using a luciferase activity assay.Results MxA protein was expressed in both the cytoplasm and nucleus of HepG2 cells,with higher expression levels in the cytoplasm than in the nucleus.Knocking down MxA expression in HepG2 cells did not affect the expression of STAT1,p-STAT1,STAT2,p-STAT2,and IRF9 proteins induced by IFN-α,but significantly reduced the expression of antiviral proteins PKR and OAS.Overexpression of MxA in HepG2 cells enhanced ISRE activity and increased the expression of PKR and OAS proteins,but this effect was inhibited in HepG2.2.15 cells.Conclusion MxA induces the expression of antiviral proteins by enhancing the activity of the JAK/STAT signaling pathway ISRE.
myxovirus resistance protein AJAK/STATinterferon-stimulated response elementinterferon-stimulated gene(ISG)
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