Abstract:[ABSTRACT] Background: MicroRNA-17 (miR-17) may participate in the regulation of human lung cancer regulation or progress. However, the detailed function and mechanisms of miR-17 are largely unknown. Objective: To construct the adenoviral vector of miR-17 and examine its roles in lung cancer cells’ proliferation, anchorage independent growth, in vitro invasion and migration. Methods: Full length sequence of miR-17 was cloned into recombinant adenovirus vectors. A549 cells were infected with adenovirus vectors (Ad-miR-17 vectors). The proliferation, anchorage-independent growth in vitro invasion or migration of A549 cells were determined by MTT-assays, soft agar-assays or trans-well assays. The specificity of miR-17 function was confirmed by transfection of miR-17’s inhibitor. Results: Infection of Ad-miR-17 significantly suppressed the proliferation, anchorage-independent growth, in vitro invasion or migration of A549 cells. Co-transfection with miR-17 inhibitor disrupted the effect of miR-17 on A549 cells. Conclusion Expression of miR-17 via identified adenoviral vector suppresses lung cancer cells proliferation.