Nitric Oxide (NO) found in the lung cancer area may play a role in regulation of cancer cell behaviors. To explore the possible effect of NO on cell motility, human lung cancer cells were exposed to non-toxic concentrations of NO for 0-14 days and migratory characteristics of the cells were determined. The present study found that long-term treatment of NO significantly enhances cell migration in dose- and time-dependent manners. Further, we found that the increased migratory action was associated with the increased levels of caveolin-1 (Cav-1) which in turn activated focal adhesion kinase (FAK) and ATP-dependent tyrosine kinase (Akt) pathways. Notably, the NO-treated cells exhibited an increased number of filopodia per cell in concomitant with an increase in cell division cycle 42 (Cdc42) proteins. Together, these results indicate a novel effect of extended NO exposure on cell migration through the Cav-1-dependent mechanism that benefits the understanding in cancer biology.