The expression levels of the protein tyrosine kinase Acki has been reported to be dysregulated in various cancers and involve in oncogenesis and progression. However, the expression and role of Ackl in osteosarcoma remains unknown. In this study, we found that Ack1 were evidently upregulated in human osteosarcoma tissues and cell lines. In addition, the clinical data showed that high expression level of Acki is closely associated with clinical stage and positive distant metastasis, and negatively correlated with overall survival. Then, bioinformatics prediction and luciferase reporter assay indicated Acki as a direct target of miR-24, and Ackl could be downregulated by miR-24 at both the mRNA and protein expression levels. Moreover, Ackl expression levels were inversely correlated with that of miR-24 in osteosarcoma tissues. Furthermore, functional assay showed that miR-24 significantly suppressed osteosarcoma progression partially mediated by inhibiting Ackl expression. Finally, western bolt assay revealed that miR-24 regulate AKT/MMPs pathway via Ackl in osteosarcoma cells. In conclusion, our study demonstrated the suppression of miR-24 on osteosarcoma metastasis by targeting Ackl via AKT/MMPs pathways, providing a novel strategy for the diagnosis and treatment of osteosarcoma patients. (C) 2017 Elsevier Inc. All rights reserved.