Background: Osteosarcoma is a common malignant tumor, with relatively lower survival rates in adolescents. Dihydrotanshinone I (DHI) was extracted from the traditional Chinese medicine Salvia miltiorrhiza and was shown to inhibit several types of cancer. Purpose: To explore the effect of DHI on the proliferation, migration, invasion, and apoptosis of osteosarcoma cells, as well as the possible molecular mechanism. Methods: The effect of DHI on the proliferation of osteosarcoma was detected by crystal violet assay, MTT assay, colony formation assay. The effects of DHI on the migration and invasion of osteosarcoma were detected by wound healing assays, cell migration and invasion assays. The effect of DHI on apoptosis of osteosarcoma was detected by cell apoptosis assay and Hoechst apoptosis staining. The protein expression levels were detected by Western blotting assay. The activity of Wnt/β-Catenin signaling pathway was detected by luciferase reporter assay and Western blot. The inhibitory effect of DHI on osteosarcoma in vivo was analyzed by an orthotopic OS tumor animal model and immunohistochemistry. Result: DHI may inhibit the proliferation, decrease the migration, reduce the invasion, and promote the apoptosis of osteosarcoma cells. In vivo mouse model, DHI can inhibit the formation of osteosarcoma. In terms of mechanism, DHI may inhibit both the transcriptional activity and the total protein level of β-catenin. Conclusion: DHI may inhibit the proliferation, migration, and invasion as well as induce the apoptosis of osteosarcoma cells, possibly through suppressing the Wnt/β-catenin signaling pathway.