This study aimed to observe the effects of puerarin on glycolysis and cisplatin sensitivity in oral squamous cell carcinoma (oSCC) cells and to explore the underlying mechanisms. CAL27 cells over- or under-expressing FBXW7 were treated with cisplatin or puerarin, and the levels of proteins involved in glycolysis as well as the activity of the respective enzymes were assessed. Glucose uptake and lactate production were also evaluated, and the IC50 value of cisplatin in CAL27 cells was determined. FBXW7 overexpression significantly downregulated HK2, PKM2, and LDH; suppressed the activity of the corresponding enzymes hexokinase, pyruvate kinase, and lactate dehydrogenase; as well as reduced glucose uptake and lactate production. FBXW7 overexpression was also associated with decreased mTOR phosphorylation and increased cisplatin sensitivity. These effects were partially antagonized by lactate or the mTOR agonist MHY1485. Puerarin suppressed glycolysis by reducing glucose uptake and lactate production, while it promoted cisplatin sensitivity and activated the FBXW7/mTOR signal pathway in a concentration-dependent manner. These effects were antagonized by FBXW7 downregulation or treatment with MHY1485. Our results suggest that FBXW7 improves cisplatin chemosensitivity and suppresses glycolysis in oSCC cells, indicating its promising potential as a target for puerarin to regulate the cisplatin sensitivity of oSCC cells.