Lysine acetylation plays an essential role in metabolism. Five individual studies have identified that a large number of cellular proteins are potentially acetylated. Notably, almost every enzyme involved in central metabolic pathways such as glycolysis, the TCA cycle, fat acid metabolism, urea cycle and glycogen metabolism, is acetylated in response to nutrition fluctuations. Metabolic reprogramming is a critical hallmark during cancer development. Tumor cells preferentially utilize glycolysis instead of oxidative phosphorylation to produce more lactate and metabolic intermediates even under normal oxygen pressure, which was first noted as the "Warburg Effect". This review focuses on recent advances in the acetylation regulation of metabolic enzymes involved in the Warburg effect, the dysfunction of acetylation regulation in tumorigenesis and their potential role in cancer metabolism therapy.