Abstract: Although multiple studies have shown that matrine can inhibit the proliferation of hepatoma cells, its mechanism of action has not been systematically investigated. In this study, the effects of matrine on the proliferation and migration of human hepatoma SMMC-7721 cells were investigated. Based on this result, anti-hepatoma target-functionally related protein interaction network of matrine was constructed, and topological analysis and clustering analysis were performed to predict the crucial targets of matrine for the anti-hepatoma effects. Pathway enrichment analysis was performed on the validated targets to predict the crucial pathways of matrine. Parts of the crucial proteins were examined by Western blot. Cellular experiments showed that matrine at concentrations of 1, 2 and 4 mg·mL^-1 significantly inhibited the proliferation of SMMC-7721 cells, and matrine at concentrations of 0.5, 1 and 2 mg·mL^-1 significantly inhibited the migration of SMMC-7721 cells. The results of network pharmacology suggest that matrine exerts its anti-hepatoma effects through acting on the key validated targets of heparanase (HPSE), caspase 3 (CASP3), Myc proto-oncogene protein (MYC), matrix metalloproteinases 2 (MMP2) and predicted targets of carbonic anhydrase 1 (CA1), lithostathine 1 alpha precursor (REG1A), carboxylesterases 1 (CES1) and acetaldehyde dehydrogenase 2 (ALDH2), and invasion and migration associated pathways. Western blot results suggest that matrine can down-regulate the expression of MMP2 and up-regulate the expression of CASP3. In this paper, we applied network pharmacology to explain the targets and pathways of matrine against hepatoma. The results provide a scientific basis for elucidation of the mechanisms of matrine against hepatoma.