Wentilactone B (EN-48-57) is one of the six derivatives separated from Aspergillus wentii (EN-48). Of these derivatives, Wentilactone B exerted a more significant antibacterial and cytotoxic activity in several tumor cell lines. The present study demonstrates that Wentilactone B could efficiently induce SMMC-7721 cells apoptosis, but not normal hepatic cells, as measured by an inverted microscope, 4',6-diamidino-2-phenylindole staining and flow cytometry. In addition, Wentilactone B could inhibit the metastasis of SMMC-7721 cells, which was detected by colony formation, scratch migration and a transwell assay, and could induce a series of intracellular events, including the down-regulation of CD44 and epidermal growth factor receptor proteins. In conclusion, Wentilactone B inhibited the growth of SMMC-7721 cells by triggering apoptosis and inhibiting metastasis.