Objective To investigate the expression level of miR-196a in non-small cell lung cancer（NSCLC） tissues and cell lines，and to explore the effect of inhibition or up-regulation of miR-196a expression on NSCLC cells proliferation and its target gene. Methods Real-time quantitative PCR was performed to detect the relative expression of miR-196a in NSCLC cell lines and tissues. MiR-196a inhibitors or mimics were transfected into SPC-A1 or A549 cells to down-regulate or up-regulate miR-196a expression, and real-time quantitative PCR was used to test the transfection efficiency. MTT and colony formation assays were performed to detect the effect of miR-196a on NSCLC cells proliferation. Bioinformatics and Western blotting analysis were performed to investigate the target genes that could be regulated by miR-196a. Results This study showed that miR-196a was highly expressed both in NSCLC samples and cell lines compared with their corresponding normal tissues and cell lines. Transfection of miR-196a inhibitors or miR-196a mimics could significantly decrease or increase its expression in SPC-A1 cells or A549 cells，respectively. Down-regulating of miR-196a expression could inhibit SPC-A1 cells proliferation by inhibiting p27kip1 expression. Conclusion Up-regulated miR-196a could promote NSCLC cells proliferation，which may affect the development of NSCLC.