Objective：To investigate the effect of miR-27b-3p on osteosarcoma cells growth and its potential molecular mechanism. Methods：①The expression of miR-27b-3p in osteosarcoma cell lines was analyzed by RT-qPCR. ②CCK-8 assay，colony formation assay and flow cytometry analysis were used to detect the effect of miR-27b-3p and structure-specific recognition protein-1(SSRP1)on osteosarcoma cell growth. ③The miRNA target prediction database was used to predict potential target gene of miR-27b-3p，and dual luciferase reporter assay was performed to demonstrate the relationship of miR-27b-3p and its target genes. ④Efficiency of knockdown or overexpression was validated by Western blot analysis. Results：①The expression of miR -27b -3p in osteosarcoma cell lines and clinical samples was low，and overexpression of miR-27b-3p inhibited the growth of osteosarcoma cells. ②Bioinformatics analysis and dual-fluorescence reporter assay confirmed that SSRP1 was a target gene of miR-27b-3p，and the expression of SSRP1 was regulated by miR-27b-3p. ③Silencing of SSRP1 significantly inhibited cell proliferation and increased cell apoptosis of osteosarcoma cells. ④Overexpression of SSRP1 partly reversed the inhibitory effect of miR-27b-3p on osteosarcoma cell growth. Conclusion：miR-27b-3p is low expressed in osteosarcoma cells，and it regulates the growth of osteosarcoma cells by targeting SSRP1. Targeting the miR-27b-3p/ SSRP1 axis may become a new therapeutic strategy for the treatment of osteosarcoma.