Tumor glycolysis is a key metabolic feature of cancer cells, characterized by increased glucose uptake and aerobic glycolysis, which produces lactate. This metabolic shift, known as the Warburg effect, is a hallmark of cancer and is linked to tumor growth, resistance to chemotherapy, and radiotherapy. Targeting glycolysis has emerged as a promising therapeutic strategy due to its role in energy production and biosynthesis of macromolecules. Recent research highlights the importance of glycolytic enzymes and their non-glycolytic functions in cancer progression. Glycolytic enzymes such as hexokinase II (HKII), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), pyruvate kinase M2 (PK-M2), and lactate dehydrogenase (LDH) play roles beyond glycolysis, influencing cell survival, proliferation, and resistance to therapy. Metabolic intermediates also contribute to non-glycolytic processes, such as the pentose phosphate pathway, which supports redox balance and biosynthesis. Targeting glycolysis through inhibitors like 3-bromopyruvate (3-BrPA) and other compounds has shown promise in preclinical studies. However, challenges remain in developing specific and effective inhibitors. Advances in targeted delivery and imaging technologies are improving the potential for glycolytic targeting in cancer therapy. Overall, targeting tumor glycolysis offers a viable approach for cancer treatment, with ongoing research aiming to translate these findings into clinical applications.Tumor glycolysis is a key metabolic feature of cancer cells, characterized by increased glucose uptake and aerobic glycolysis, which produces lactate. This metabolic shift, known as the Warburg effect, is a hallmark of cancer and is linked to tumor growth, resistance to chemotherapy, and radiotherapy. Targeting glycolysis has emerged as a promising therapeutic strategy due to its role in energy production and biosynthesis of macromolecules. Recent research highlights the importance of glycolytic enzymes and their non-glycolytic functions in cancer progression. Glycolytic enzymes such as hexokinase II (HKII), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), pyruvate kinase M2 (PK-M2), and lactate dehydrogenase (LDH) play roles beyond glycolysis, influencing cell survival, proliferation, and resistance to therapy. Metabolic intermediates also contribute to non-glycolytic processes, such as the pentose phosphate pathway, which supports redox balance and biosynthesis. Targeting glycolysis through inhibitors like 3-bromopyruvate (3-BrPA) and other compounds has shown promise in preclinical studies. However, challenges remain in developing specific and effective inhibitors. Advances in targeted delivery and imaging technologies are improving the potential for glycolytic targeting in cancer therapy. Overall, targeting tumor glycolysis offers a viable approach for cancer treatment, with ongoing research aiming to translate these findings into clinical applications.