Propofol is a commonly used anesthetic drug with potential antitumor activity. The aim of this study was to investigate the antiproliferation and antimetastatic mechanisms of propofol in hepatocellular carcinoma (HCC). SMMC-7721 was treated with different concentrations of propofol. Cell proliferation was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and cell motility was assessed by the wound healing assay. Cell migration and invasion ability were analyzed by the transwell assay. Protein levels of E-cadherin, Vimentin, matrix metalloproteinase 2 (MMP-2), and MMP-9 were measured by Western blotting. Twist1 gene expression was assessed by Western blotting and quantitative reverse transcription PCR. The proliferation, motility, migration, and invasion of SMMC-7721 cells were inhibited by propofol treatment in a dose-dependent manner. Propofol treatment downregulated the protein levels of Vimentin, MMP-2, and MMP-9 while increasing that of E-cadherin. Propofol treatment inhibited the expression of the Twist1 gene in SMMC-7721 cells. The overexpression of Twist1 could partially reverse the inhibitory effects of propofol on SMMC-7721 cells. Propofol inhibited proliferation, migration, and invasion of HCC cells by downregulating the expression of Twist1, making it a potential drug for HCC treatment.