Here, for the first time, we reported that neddylation inhibition with MLN4924 induces the accumulation of ATF3 to trigger pro-survival autophagy by modulating NF-κB-Catalase-ROS-ATF3 axis in esophageal cancer cells, highlighting targeting ATF3-mediated autophagy as a potential strategy to enhance neddylation-targeted anti-ESCC therapy. Here, CAT is linked to esophageal squamous cell carcinoma.