An in-depth analysis of SPDEF-targeted genes revealed four crucial genes for the advanced prognostic model, among which S100A16 was significantly correlated with SPDEF. Mechanistic analysis showed that SPDEF enhances the transcription of S100A16, which in turn enhances PAAD cell migration, proliferation, and invasion by activating the PI3K/AKT signaling pathway. Here, S100A16 is linked to pancreatic adenocarcinoma.