Background The present study explored the role and mechanism of microRNA-874-3p (miR-874-3p) in the migration of the osteosarcoma cell line, U-2 OS. Methods The expression profile of osteosarcoma (OS) microRNA (GSE65071) datasets was downloaded from the Gene Expression Omnibus (GEO) database () to identify differentially expressed miRNAs in OS and its biological functions. A quantitative reverse transcription-polymerase chain reaction was performed to detect the expression of miR-874-3p and its target gene regulator of G protein 4 (RGS4) in human osteosarcoma cells U-2 OS and normal osteoblast hFOB1.19. Plasmid overexpression miR-874-3p and pcDNA-RGS4 were transfected into U-2 OS using Lipofectamine 2000 (Thermo Fisher, Waltham, MA, USA). Cell migration was measured using Transwell migration assays. Bioinformatic analysis and luciferase reporter assay were conducted to search for the target gene of miR-874-3p. Results In total, 167 differentially expressed miRNAs were detected after the analysis of GSE65071; of which 78 were up-regulated genes and 89 were down-regulated. miR-874-3p was down-regulated and selected for further analysis. The expression level of miR-874-3p in U-2 OS cells was significantly decreased compared to the hFOB1.19 cell line (p < 0.05). Overexpression of miR-874-3p significantly inhibited the proliferation and migration of U-2 OS cells and overexpression of RGS4 reversed the inhibitory effect of miR-874-3p on U-2 OS cells. Through luciferase report analyses and bioinformatic analysis, RGS4 may be the candidate target gene of miR-874-3p. Conclusions In conclusion, overexpression of miR-874-3p suppressed OS cell proliferation and migration. Thus, miR-874-3p might present a therapeutic agent for the treatment of OS.