In order to assess whether miR-16 is able to overcome the TKIs’ primary resistance by targeting MAPK3, MAP2K1, and CRAF located downstream of the KRAS-activated oncogene, we transfected KRAS-amplified NSCLC cell lines A549 and NCI-H2009 with the pre-miR-16 mimic or a scrambled miRNA as the control and determined the in vitro cell growth after treatment with erlotinib 1 μg/mL for 6 h, followed by a 48–72–96-h washout. This evidence concerns the gene MAP2K1 and non-small cell lung carcinoma.