Aim: To provide experimental data for further research on the signal transduction of apoptosis in lung adenocarcinoma cells, we examined the effects of exogenous C2-ceramide administration on several members of the mitogen-activated protein kinase (MAPK) superfamily and caspase-3 in A549 cells. Methods: Cell viability and apoptosis were analyzed by cell counting kit-8 assay and flow cytometry.Various MAPK and caspase-3 proteins were detected by Weste blotting.Results: C2-ceramide selectively altered the phosphorylation state of menbers of the MAPK superfamily, causing hyperphosphorylation ofmitogen-activated pro-tein kinase kinase (MEK)1/2 and the p38 MAPK, but not affecting the phosphory-lation of extracellular signal-regulated kinase 1/2 and the c-Jun N-terminal kinase.SB-203580 (a p38 MAPK inhibitor) and p38 siRNA, but not U0126 (a MEK inhibitor),partially rescued cell death induced by C2-ceramide. C2-ceramide promoted the activation of caspase-3. Conclusion: Exogenous C2-ceramide induced apoptosis in human lung adenocarcinoma A549 cells. The activation of MAPK and caspase-3 were involved in the mechanisms of C2-ceramide-induced apoptosis in A549 cells.