Cis‑diamminedichloroplatinum (cisplatin, CDDP)‑containing combination chemotherapy is commonly used for the treatment of non‑small‑cell lung cancer (NSCLC). 2',2'‑Difluorodeoxycytidine (gemcitabine, GEM), an active antineoplastic agent for NSCLC, has been previously reported to be suitable for use in combination with cisplatin in chemotherapy, since their mechanisms may be complementary. In the present study, the sequence‑dependent effects of GEM and CDDP were investigated in the NSCLC cell line, A549. Significantly increased rates of inhibition and cell cycle arrest were observed in the group treated with GEM followed by CDDP, and this treatment plan was demonstrated to represent the most efficient treatment protocol for the A549 NSCLC cell line. Results of the present study are consistent with previous studies in other cell lines and are likely to provide important insight for subsequent studies.