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OBJECTIVE To investigate the antiproliferative and apoptogenic activities of peptide extracted from the Chinese toad (Bufo bufo gargarizans) skin (TSP) and its effects on hepatocarcinoma cell line. METHODS MTT assaywas used to detect the effects of TSP (50 mg/mL and 5 mg/mL) on the proliferation and viability of Hepatocarcinoma cell line (HepG2) and liver cell line (L-02); Flow cytometry was used in DNA content analysis to determine the cell distribution in different phases of cell cycle; Annexin V-FITC/PI stained fluorescence-activated cell sorter (FACS) and transmission electron microscope (TEM) were used to detect the apoptosis of the treated cells. RESULTS TSP could not suppress the proliferation and viability of normal liver L-02 cells, but strongly inhibited the proliferation and viability of HepG2 cells; TSP (50 mg/mL) primarily arrested the HepG2 cells at G1 phase of the cell cycle; TSP (50 mg/mL) induced apoptosis in HepG2 cells and enhanced the effects of 5-Fu. CONCLUSION TSP has potent antineoplastic activity against human hepatocarcinoma cells with little toxicity to normal liver cells and can enhance the effects of 5-Fu.