Objective To investigate the effects of silencing eukaryotic translation initiation factor 5A-2 (eIF5A2) on drug resistance and autophagy of human lung cancer cell line A549/DDP. Methods Human lung cancer cell lines A549 and A549/DDP were cultured in vitro. The A549/DDP cells were transfected, and divided into eIF5A-2-SiRNA group, negative control group and blank control group, A549 cells were used as the control group. The expression level of eIF5A2 mRNA in cells was detected by realtime fluorescent quantitative PCR (qRTPCR). The drug resistance of A549/DDP cells was detected by MTT. The apoptosis rate was detected by flow cytometry. The autophagy was detected by monodansyl cadaverine (MDC) staining. The relative expression levels of eIF5A-2, LC3Ⅱ and Beclin1 proteins were detected by Western blot. Results Compared with the normal control group, the relative expression levels of eIF5A-2 mRNA in A549/DDP cells in the blank control group and NC group were significantly higher (P＜0.05). Compared with the control group and NC group, the apoptosis rate of eIF5A-2-SiRNA group was significantly higher (P＜0.05), the IC50, number of autophagy bodies, relative expression levels of eIF5A2, LC3Ⅱ and Beclin1 proteins decreased significantly (P＜0.05). Conclusion T he down-regulation of eIF5A-2 can reduce the drug resistance of human lung cancer cell line A549/DDP, which may be related to the downregulation of LC3Ⅱ and Beclin-1 expressions and the inhibition of autophagy.