c-Jun is an AP-1 transcription factor and implicated in many aspects of cellular functions, but its exact role in the regulation of early intestinal epithelial restitution after injury remains largely unknown. Phospholipase C-1 (PLC1) catalyzes hydrolysis of phosphatidylinositol 4,5 biphosphate into the second messenger diacylglycerol and inositol 1,4,5 triphosphate, and coordinates Ca²⁺ stores mobilization, and regulates cell migration and proliferation in response to stress. Here we reported that c-Jun up-regulates PLC1 expression and enhances PLC1-induced Ca²⁺ signaling, thus promoting intestinal epithelial restitution after wounding. Ectopically expressed c-Jun increased PLC1 expression at the transcription level and this stimulation is mediated by directly interacting with AP-1- and C/EBP-binding sites that are located at proximal region of the PLC1 promoter. Increased levels of PLC1 by c-Jun elevated cytosolic free Ca²⁺ concentration and stimulated intestinal epithelial cell migration over the denuded area after wounding. The c-Jun-mediated PLC1/Ca²⁺ signal also plays an important role in polyamine-induced cell migration after wounding, since increased c-Jun rescued Ca²⁺ influx and cell migration in polyamine-deficient cells. These findings indicate that c-Jun induces PLC1 expression transcriptionally and enhances rapid epithelial restitution after injury by activating Ca²⁺ signal.