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Hepatocellular carcinoma(HCC)is one of the most frequent types of malignant tumors in the world.Traditional chemotherapy and radiotherapy have cytotoxic effects on normal cells.This cytotoxicity coupled with severe toxicity highlight the limitations of traditional therapies and illustrates the urgent need for safer and more selective treatment options with fewer side effects in HCC therapy.Complementary and alternative medical therapies belonging to traditional Chinese medicine(TCM)have demonstrated clinically effectiveness in cancer treatment.Hedyotis Diffusa Willd formula(HDWF)is a four-herb Chinese medicinal Medicine Formula.To elucidate the anticancer effects of HDWF,MTT assay,colony formation,nuclear staining,andAnnexin-V/PI staining assays were used to evaluate the effect of HDWF on cell proliferation and apoptosis of HCC cell lines(SK-Hep-1 and Bel-7402).Furthermore,western blotting was performed to detect the expression of some apoptosis associated proteins.The results showed HDWF reduced HCC cell viability and attenuated cell proliferation in a dose-and time-dependent manner.Moreover,HDWF increased the expression of Fas,FasL,and Bax,downregulated the expression of Bcl-2 and promoted the activation of caspase-8,-9,and-3.These results suggested HDWF possessed anticancer effects and the mechanisms may involve the mitochondrial apoptosis pathway and death receptor pathway.