objective: To investigate the role of microRNA-582-5p (miR-582-5p) on osteosarcoma (OS) cell behaviors and provide potential therapeutic targets for OS. methods: Expression levels of miR-582-5p in OS cell lines and normal cell lines were measured by quantitative real-time PCR. Cell proliferation, migration and invasion capacities were assessed by cell counting kit-8 assay, wound-healing assay, and transwell invasion assay, respectively. Downstream target of miR-582-5p was confirmed by Luciferase activity reporter assay and Western blotting assay. results: MiR-582-5p expression was downregulated in OS cell lines compared with normal cell lines. Overexpression of miR-582-5p inhibits OS cell proliferation, migration, and invasion capacities. Neuro-oncological ventral antigen 1 (NOVA1) was chosen as target gene of miR-582-5p by bioinformatics analysis and Luciferase reporter assay. Moreover, overexpression of NOVA1 could impair tumor suppression role of miR-582-5p on OS cell behaviors. conclusions: MiR-582-5p exerts tumor-suppressive role on OS cell behaviors via targeting NOVA1 in vitro, which will help us to understand the mechanisms underlying OS progression.