Current chemotherapeutic strategies for liver cancer have limitations. Thus, the demand for complementary medicine is warranted. We evaluated the antitumor potential of berberine, a naturally bioactive phytochemical from Coptis chinensis Franch against Huh7 cancer cells and WRL68 liver cells. The antitumor activity of berberine was evaluated by flow cytometry. The caspase-dependent pathway was assessed using western blot analysis. Results showed that berberine induced the apoptosis of liver cancer cells through procaspase-9, and its effector caspases, procaspase-3 and procaspase-7. Flow cytometry revealed that berberine caused cell cycle arrest at the M/G1 phase. The results of reverse transcription-polymerase chain reaction showed that berberine increased the expression of Bax, which resulted in the activation of the caspase cascade. The present findings demonstrated that berberine induces the apoptosis of Huh7 cells via the mitochondrial pathway.