Objective To investigate the effects of rapamycin on the growth and proliferation of human non-small cell lung cancer cell line A549. Methods A549 cells were cultured with 0 (negative control group), 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, 1 and 5 μmol/L rapamycin (rapamycin intervention group) for 24 h, 48 h and 72 h, and cell viability was detected by MTT assay. A549 cells were cultured with 0 (negative control group), 0.01, 0.05, 0.1, 0.5 and 1 μmol/L rapamycin (rapamycin intervention group) for 48 h, cell morphology was observed by light microscopy, and cell cycle was analysed by flow cytometry with PI simple staining. Results MTT assay revealed that the cell growth inhibition rate in rapamycin intervention group was significantly higher than that in negative control group (P<0.05), and the cell growth inhibition rate was most significant after culture with 5 μmol/L rapamycin for 72 h. Compared with negative control group, A549 cells tended to maintain a round shape by contracting the pseudopod, and the gaps between cells enlarged after culture with different concentrations of rapamycin for 48 h. Cell cycle detection indicated that compared with negative control group, the percent of cells in G0/G1 phase significantly increased, and the percent of cells in S phase significantly decreased in rapamycin intervention group (P<0.05). Conclusion Rapamycin has significant inhibitory effects on the growth and proliferation of human lung cancer cell line A549 in a time- and concentration-dependent manner.