This study is to investigate the effect and its possible mechanisms of lidamycin (LDM) combined with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in human non-small cell lung cancer (NSCLC) cells.  MTT assay was used to determine the growth inhibition of the two ingredients on H460 cells.  Apoptosis was examined by Annexin V-FITC/PI staining, flow cytometry assay and DNA-specific dye Hoechst 33342 staining. The level of TRAIL receptor and apoptosis-associated protein expression was detected by  Western blotting analysis.  The results showed that the IC₅₀ value of LDM and TRAIL for H460 cells was 4.603×10⁻¹⁰ mol·L⁻¹ and 915.3 ng·mL⁻¹ respectively, but the IC₅₀ value of LDM was 3.064×10⁻¹¹ mol·L⁻¹ and 1.611×10⁻¹¹ mol·L⁻¹ when different concentrations of LDM was combined with 50 and 100 ng·mL⁻¹ TRAIL   respectively.  And the CDI value was less than 1.  The apoptosis ratios also increased in the combination group relative to the single-agent treatment and the untreated control.  Furthermore, the induction of the cleavage of PARP and the activation of Caspase-3 and Caspase-8 by the combination were more effective than LDM or TRAIL alone.  At last, the level of death receptor 5 (DR5) expressions increased in a dose-dependent manner and time-related pattern.  The data indicate that LDM inhibits the growth of H460 cells in vitro.  DR5 induction contributes to enhancement of TRAIL-induced apoptosis by LDM in human non-small lung cancer cells.