Prostaglandin E2 (PGE2), a major product of cyclooxygenase-2 (COX-2), plays an important role in the carcinogenesis of many solid tumors including colorectal cancer. Because PGE2 functions by signaling through PGE2 receptors (EPs), which regulate tumor cell growth, invasion, and migration, there is growing interest in the therapeutic potential of targeting EPs. In the present study, we investigated the role of EP4 on the effectiveness of cordycepin in inhibiting migration and invasion of HCT116 human colorectal carcinoma cells. Our data indicate that cordycepin suppressed lipopolysaccharide (LPS)-enhanced cell migration and invasion through inactivation of matrix metalloproteinase (MMP)-9 and down-regulation of COX-2 expression and PGE2 production. These events were associated with inactivation of EP4 and activation of AMP-activated protein kinase (AMPK). Moreover, the EP4 antagonist AH23848 prevented LPS-induced MMP-9 expression and cell invasion in HCT116 cells. However, the AMPK inhibitor, compound C, and knockdown of AMPK activity via siRNA attenuated the cordycepin-induced inhibition of EP4 expression. Cordycepin treatment also reduced the transcriptional activity of cAMP responsive element-binding protein (CREB). These findings provide evidence that cordycepin suppresses the migration and invasion of HCT116 cells through modulating EP4 expression and the AMPK-CREB signaling pathway. Therefore, cordycepin has the potential to be a potent anti-cancer agent in therapeutic strategies against colorectal cancer metastasis.