Objective:To explore the role of PI3K/Akt signal pathway in acetylation and aerobic glycolysis in Huh-7 cells. Methods:We treated Huh-7 cells with LY294002 (PI3K/Akt inhibitor) and rapamycin (mTOR inhibitor). DMSO treatment was set as negative control. Lactate concentrations were measured by lactate kits. The expression of ACL and acetylation was measured by Western blots. Cell viability was determined by WST assay. Results:①Under LY294002,lactate concentrations decreased by 66.15%. Expression levels of ACL declined by 20.84%. Acetylation levels decreased at several sites(P < 0.01,n = 3). ②Under rapamycin,lactate concentrations decreased significantly by 90.16%. Expression levels of ACL declined by 64.16%. Acetylation levels were down-regulated variously at different sites (P < 0.01,n =3 ). ③After 24 h culture with 20mmol/L LY294002 or 20 μg/L rapamycin,the cell growth inhibition ratio reached to 24.78% and 20.67%,respectively(P < 0.01,n = 3). Conclusion:PI3K/Akt/mTOR signal pathway promotes the aerobic glycolysis of cancer cells by up-regulating the expression of ACL genes and the levels of acetylation,which may contribute to the tumor growth.