The treatment of acute myeloid leukemia (AML) is a major clinical challenge, with patients often having poor prognoses, especially in subtypes with high-risk genetic profiles. PGAM5 plays a critical role in the progression of various malignancies; however, its biological function and underlying molecular mechanisms in AML remain unclear. The study aimed to systematically investigate the role and potential mechanisms of PGAM5 in AML. Bioinformatics analysis revealed that PGAM5 expression was significantly upregulated in AML patients compared with healthy controls, and high PGAM5 expression was closely associated with unfavorable prognosis. Further analysis suggested that PGAM5 may be related to the PI3K/AKT/mTOR signaling pathway. By establishing AML cell models with PGAM5 knockdown, functional experiments demonstrated that suppressing PGAM5 expression significantly arrested cell cycle progression, inhibited proliferation, and induced apoptosis. Mechanistic studies indicated that PGAM5 is closely associated with glycolytic metabolism in AML and enhances glycolytic flux through the transcription factor HIF-1α. Upon PGAM5 knockdown, AML cells exhibited significant reductions in glucose consumption, ATP production, and lactate output. Furthermore, treatment with the PI3K activator 740Y-P in PGAM5-knockdown cells provided additional evidence that PGAM5 plays a critical role in supporting AML cell proliferation and metabolic reprogramming, suggesting that PGAM5 may represent a candidate molecule for further functional investigation in AML. In summary, this study elucidates the key role of PGAM5 in metabolic remodeling in AML, suggesting its potential as a novel therapeutic target for AML treatment.