Solasonine (SS) is a natural glycoalkaloid compound that has been reported to possess a significant anticancer function. However, its anticancer effects and related mechanisms in osteosarcoma (OS) have not been studied. This study sought to investigate the impact of SS on the growth of OS cells. OS cells were treated with different concentrations of SS for 24[Formula: see text]h, and the results showed that SS attenuated the survival of OS cells in a dose-dependent manner. Additionally, SS suppressed cancer stem-like properties and epithelial-mesenchymal transition (EMT) by inhibiting aerobic glycolysis in OS cells in an ALDOA-dependent manner. Additionally, SS reduced the levels of Wnt3a, [Formula: see text]-catenin, and Snail in OS cells in vitro . Furthermore, Wnt3a activation reversed the SS-induced inhibition of glycolysis in OS cells. Collectively, this study discovered a novel effect of SS in inhibiting aerobic glycolysis, in addition to cancer stem-like features and EMT, implying that SS could be a therapeutic candidate for OS treatment.