Glycolysis plays a critical role in tumor development and progression by promoting cell proliferation, invasion, metastasis, and immune evasion. Tumor cells exhibit enhanced glycolytic activity, leading to increased lactate production and an acidic tumor microenvironment (TME), which supports tumor growth and suppresses immune responses. Glycolytic enzymes and products serve as potential biomarkers and therapeutic targets for tumor treatment. Recent studies have shown that glycolytic enzymes such as HK2, PKM2, and ENO1 are upregulated in various cancers and are involved in tumor progression. Inhibitors of glycolytic enzymes, including 2-deoxy-D-glucose (2-DG), 3-bromopyruvate (3-BrPA), and metformin, have been developed to target tumor cells and enhance chemotherapy sensitivity. Additionally, glycolysis is closely linked to tumor angiogenesis, which facilitates tumor growth and metastasis. The regulation of glycolysis in tumor cells provides new insights for the diagnosis and treatment of tumors, highlighting the importance of targeting glycolytic enzymes in cancer therapy.Glycolysis plays a critical role in tumor development and progression by promoting cell proliferation, invasion, metastasis, and immune evasion. Tumor cells exhibit enhanced glycolytic activity, leading to increased lactate production and an acidic tumor microenvironment (TME), which supports tumor growth and suppresses immune responses. Glycolytic enzymes and products serve as potential biomarkers and therapeutic targets for tumor treatment. Recent studies have shown that glycolytic enzymes such as HK2, PKM2, and ENO1 are upregulated in various cancers and are involved in tumor progression. Inhibitors of glycolytic enzymes, including 2-deoxy-D-glucose (2-DG), 3-bromopyruvate (3-BrPA), and metformin, have been developed to target tumor cells and enhance chemotherapy sensitivity. Additionally, glycolysis is closely linked to tumor angiogenesis, which facilitates tumor growth and metastasis. The regulation of glycolysis in tumor cells provides new insights for the diagnosis and treatment of tumors, highlighting the importance of targeting glycolytic enzymes in cancer therapy.