In this study, based on the discovery of an interaction between TRAF6 and IRF3, we further expanded their contribution to the nuclear translocation of NF-κB-p65, that is, the TRAF6-IRF3 axis induced the activation of NF-κB-p65 and finally promoted chemoresistance of GC cells to 5-FU. This evidence concerns the gene IRF3 and gastric cancer.