Non-small cell lung cancer (NSCLC) is the most lethal malignant tumor and is also considered one of the most chemoresistant cancers. Despite the benefits obtained from platinum-based therapy, the majority of patients treated will progress and die. In the continuing quest for personalized therapy based on the biomolecular characteristics of each single patient, clinical practice now seems to be oriented towards combining conventional drugs with molecular-targeted agents. In the present study, we evaluated the antitumor activity of docetaxel, one of the most widely used drugs for second-line treatment, and Iressa, an EGFR-targeting tyrosine kinase inhibitor, administered singly or in sequence. The study was performed on three human NSCLC cell lines (ChaGo-K1, CAEP and RAL) that exhibit different expression of proliferation and apoptosis-related markers, and do not harbor EGFR mutations. The efficacy of docetaxel and Iressa differed in the three cell lines and an important synergistic interaction was observed with the sequence 1-h docetaxel --> 72-h Iressa during which Iressa doubled the fraction of docetaxel-induced apoptotic cells, amplifying a caspase-dependent apoptosis and inhibiting docetaxel-induced p21 hyperexpression. Moreover, the important role of MAPK-dependent modulation of this molecular marker was shown using a specific inhibitor. The results from the present preclinical study demonstrate the cytotoxic activity of Iressa and its ability to increase taxane activity in a model that does not harbor EGFR-specific mutations, thus highlighting the importance of focusing on alternative molecular targets of Iressa activity.