Most cancer-related deaths are caused by the development of metastatic disease. Thus, investigation of the underlying mechanisms of metastasis is urgent to design more effective targeted drugs and to treat metastatic disease more effectively. MicroRNAs (miRNAs) have emerged as potential targets for cancer treatment. In the present study, we aimed to identify the roles of miR-134 in non-small cell lung cancer (NSCLC) cell migration and invasion. We demonstrated that overexpression of miR-134 inhibited migration and invasion of A549 and H1299 cells. Further mechanistic investigations revealed that miR-134 inhibited epithelial-to-mesenchymal transition (EMT) evidenced by upregulation of E-cadherin expression and downregulation of vimentin expression. Using luciferase assays, we identified integrin β1 (ITGB1) as a direct target of miR-134. Performing RNAi and rescue experiments, we confirmed that miR-134 exerted its migratory and invasive suppressive role partly by downregulating ITGB1. Finally, an in vivo experiment also, to some extent, suggested that miR-134 may function as a suppressor of metastasis. Taken together, our findings suggest that miR-134 suppresses migration and invasion of NSCLC by targeting ITGB1.