Objective To observe the anti-cancer effect in vitro of Aitongxiao Granules containing serum on SMMC-7721 and Bel-7404 liver cancer cells, thus revealing the anti-cancer mechanism of the treatment in adjusting proliferation and apoptosis. Methods Male Sprague-Dawley rats were administered by gastrogavage Aitongxiao decoction of 86.4, 43.2, 21.6 g/kg twice a day for 1 week. Using serum pharmacologic method, the proliferation of SMMC-7721 and Bel-7404 liver cancer cells were determined by MTT chromatometry after co-cultured with medicated serum containing different concentration of Aitongxiao decoction. The apoptosis of SMMC-7721 and Bel-7404 liver cancer cells were detected by flow cytometry. Results The research in vitro showed that Aitongxiao Granules containing serum low, medium and high dose groups had varying degrees of inhibition on hepatoma cells, and this inhibition showed a concentration dependence within a certain range. Aitongxiao Granules containing serum could induce apoptosis of human hepatoma SMMC-7721 cells and Bel-7404 cells. Conclusion Aitongxiao Granules showed effect of inhibiting proliferation and inducing apoptosis on SMMC-7721 and Bel-7404 cells in vitro.