Abstract: The aim of this study was to investigate the effect of β-elemonic acid（β-EA） on hepatocellular carcinoma（HCC） proliferation and apoptosis and its mechanism.CCK8 assay was used to detect the effects of β-EA on the cell viability of human HCC cells Huh7 and PLC/PRF/5. Annexin V-FTTC/7AAD staining was used to detect cell apoptosis. The effect of β-EA on cell cycle of HCC cells was detected by flow cytometry. The expressions of apoptosis-related proteins Bcl2, BAX, cleaved-caspase 3 and cell cycle related proteins CDK1 and Cyclin B1 were detected by Western blot. The molecular mechanism of β-EA inhibiting the proliferation of HCC cells was predicted by network pharmacology, and the protein expression of PI3K/AKT signaling pathway was detected. The growth inhibition effect of β-EA on HCC was confirmed by constructing H22 subcutaneous transplantation tumor model in vivo. The results showed that β-EA significantly inhibited Huh7 and PLC/PRF/5 cell viability, induced cell apoptosis and G2/M cell arrest. The expression of pro-apoptotic protein cleaved-caspase 3 and BAX was up-regulated, and the expression of anti-apoptotic protein Bcl2, cell cycle protein Cyclin B1, cycle-dependent kinases CDK1, p-PI3K, and p-AKT was down-regulated. The subcutaneous tumor transplantation model confirmed that β-EA can effectively reduce tumor weight, tumor volume, and Ki67 expression. The results of this study suggest that β-EA can induce apoptosis and G2/M cell arrest of HCC cells through PI3K/AKT signaling pathway.