In conclusion, the current study illustrates that N-nitrosamine-mediated upregulation of FadAL induces the formation of EMT subtypes in ESCC by directly interacting with FLOT1 and promoting PA accumulation, thereby enhancing Wnt3A palmitoylation and activating the Wnt/β-catenin signaling pathway. Here, FLOT1 is linked to esophageal squamous cell carcinoma.