Abstract: Objective To investigate the effects of IWR-1 on proliferation and the possible mechanism of human hepatic carcinoma cell lines Hep3B. Methods Hep3B cells were treated with different concentration of IWR-1(2,4,8,16μmol/L). Cell growth inhibition rates were determined by CKK-8 assay. Cell cycle and apoptosis rate were analyzed by flow cytometry(FCM). The expression levels of proteins were detected by Western blotting. β-catenin and c-myc mRNA expression were determined by real time polymerase chain reaction(RTPCR). Results The proliferation rate of Hep3B cells in the IWR-1 treated group decreased in a time- and concentration- dependent manner(P<0.01). FCM results showed that the proportion of cells in G0/G1 phase were increased while the apoptosis rate of Hep3B cells was decreased in a concentration-dependent manner(P<0.01). IWR-1 could down-regulate β-catenin and c-myc mRNA levels, down-regulate β-catenin and c-myc mRNA protein levels, but significantly up-regulate Axin 1 and p-β-catenin protein levels. Conclusion IWR-1 can inhibite proliferation of Hep3B cells through inhibiting Wnt/β-catenin signaling pathway.