Objective: To investigate the effect of osthole on the proliferation and invasion of hepatocellular carcinoma cells HepG2, SMMC-7721, Bel-7402. Methods: Human liver cancer cells were taken for resuscitation,passage and culture. The experimental setting was divided into a blank group and an administration group. The administration group was treated with different concentrations of osthol liquid (50, 100, 150, 200 μmol/L). MTT method and cell colony experiment were used to determine the inhibitory effect of different concentrations of osthole on the proliferation of human liver cancer cells in vitro. Wound healing assay and transwell assay were used to investigate the inhibition of migration and invasion on HCC. Western blot was used to detect the influence of E-cadherin expression, a protein related to transformation (EMT) and MMP-2, a protein related to cell migration were detected by Western blot. Results: Osthole could inhibit the proliferation of HepG2, SMMC-77721 and Bel-7402 in vitro, with IC50 values of 147.62, 141.57 and 179.67 μmol/L, respectively, showing a certain concentrationdependent. Compared with the blank group, the treatment group could significantly inhibit the proliferation of human hepatocellular carcinoma cells under the conditions of 50, 75, and 100 μmol/L (P<0.05). It was found that osthole could effectively inhibit the migration and invasion of HCC. Compared with the blank group, the administration group showed statistically significant difference (P<0.05) The protein expression of MMP in the cells of the administration group decreased (P<0.05), while the expression of E-cadherin protein increased (P<0.05). Conclusion: Osthole can inhibit the proliferation and invasion in HCC and its mechanism may be related to protein E-cadherin and MMP-2.