By establishing HNSCC xenograft mice models and metastatic mice models, we further tested the therapeutic efficacy of PFK15 on HNSCC in vivo, and proved that the blockage of glycolysis by targeting PFKFB3 not only suppressed tumor growth but also alleviated the distant metastasis of HNSCC, reflecting the promising application of PFK15 for HNSCC treatment. The gene discussed is PFKFB3; the disease is head and neck squamous cell carcinoma.