Revisiting the Warburg effect: moonlighting functions of glycolytic-related enzymes in tumorigenesis

Tumor cells exhibit distinct metabolic reprogramming features, among which aerobic glycolysis (Warburg effect) is one of the most prominent characteristics. Traditionally, metabolic enzymes were considered to function primarily in catalyzing biochemical reactions; however, recent studies have revealed that, beyond their metabolic roles, metabolic enzymes also play critical roles in tumorigenesis and progression through non-enzymatic mechanisms. This review systematically elaborates on the molecular mechanisms by which glycolytic enzymes and related pathway enzymes—such as HK2, ENO1, and PKM2—participate in tumor progression and immune evasion via non-enzymatic activities, including transcriptional regulation, RNA binding, organellar translocation, secretion, and protein-protein interactions. Furthermore, we discuss the implications of these findings for targeted cancer therapy and propose clinical intervention strategies targeting the non-enzymatic functions of these metabolic enzymes.