Objective: To establish apoptosis model of cultured lung cancer cells. Methods: A549 human lung adenocarcinoma cells were treated in cultured fluid containing 20 μmol/L doxorubicin for 4 h. At different time points of post-incubation the detached cells were harvested for analysis by counting, trypan blue exclusion, acridine orange dying, electromicroscopy, DNA agarose gel electrophoresis and flow cytometry. Results: The morphology of the treated cells was characterized by positive trypan blue exclusion, membrane budding, cytoplasmic condensation, chromatin condensation, fragmentation, margination and apoptotic body formation. Agarose gel electrophoresis analysis revealed DNA cleavage (called DNA ladder). An apoptotic peak appeared in the analysis of flow cytometry. All those results suggested that apoptosis of A549 cells was successfully induced by doxorubicin. Most of the apoptotic cells appeared in the detached cell group after 10-12 h of post-incubation. Conclusion: Apoptosis of A549 lung cancer cells can be induced by doxorubicin.