Abstract: Objective To investigate the inhibitory effect of polysaccharide of Atractylodes macrocephala (PAM) on the proliferation and invasion of hepatocellular carcinoma cells and the underlying mechanism. Methods Hepatocellular carcinoma HepG2 cells were treated with different concentrations of PAM, and their proliferation and invasive ability were examined using CCK-8 assay and Transwell assay. Immunofluorescence assay was performed to detect the expression level of β-catenin, and real-time PCR and Western blotting were used to detect the mRNA and protein expressions of AKT, GSK-3β and MMP-2 in the cells. The changes in the proliferation, invasiveness and the expressions of pGSK-3β and MMP2 were examined in the cells following treatment with LiCl/PAM/LiCl plus PAM. Results PAM treatment significantly reduced the cell viability, the number of migration cells, and the expression levels of β-catenin and MMP-2 (P<0.05), and obviously inhibited the phosphorylation of AKT and GSK-3β in the cells (P<0.05) in a dose-dependent manner. The rescue experiment showed that LiCl reversed the inhibition of cell proliferation, invasiveness, and the Wnt/β-catenin pathway induced by PAM. Conclusion PAM can inhibit the proliferation and invasion of hepatocellular carcinoma cells in vitro possibly by inhibiting the Wnt/β-catenin signaling pathway.