Cinobufacini, an aqueous extract from Bufo bufo gargarizans Cantor, induces apoptosis through a mitochondria-mediated pathway in human hepatocellular carcinoma cells

Aim of the studyCinobufacini (Huachansu), an aqueous extract from the skin and parotid venom glands of Bufo bufo gargarizans Cantor, is a traditional Chinese medicine widely used in clinical cancer therapy in China. The present study sought to investigate the possible signaling pathway implicated in cinobufacini-induced apoptosis in the hepatocellular carcinoma cell lines HepG2 and Bel-7402.Materials and methodsThe effects of cinobufacini on cell proliferation of HepG2 and Bel-7402 cells were evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assays. Cell apoptosis was detected by Hoechst 33258 staining and flow cytometry analysis. The mitochondrial membrane potential (Δψm) and caspase-9 and -3 activity were detected using MitoCapture reagent staining and colorimetric assays, respectively. The expression of apoptosis-related proteins and release of cytochrome c were assessed by Western blot analysis.ResultsCinobufacini significantly inhibited cell proliferation of both cell lines in a dose- and time-dependent manner. Marked changes in apoptotic morphology and apoptosis rates were clearly observed after cinobufacini treatment. The protein expression of Bax increased whereas that of Bcl-2 decreased, leading to an increase in the Bax/Bcl-2 ratio. Subsequently, cinobufacini disrupted the mitochondrial membrane potential (Δψm) and resulted in the release of cytochrome c, activation of both caspase-9 and -3, and cleavage of poly (ADP-ribose) polymerase (PARP).ConclusionThe present study indicated that cinobufacini can induce apoptosis of HepG2 and Bel-7402 cells through a mitochondria-mediated apoptosis pathway.

Cinobufacini (Huachansu) is an aqueous extract from the skins and parotid venom glands of the toad Bufo bufo gargarizans Cantor (shown in the figure is the dried skin of Bufo bufo gargarizans Cantor) and is a traditional Chinese medicine that is widely used in clinical cancer therapy in China. This study mainly investigated the apoptosis-inducing effects of cinobufacini and its possible signaling pathway in the hepatocellular carcinoma cell lines HepG2 and Bel-7402.Figure optionsDownload as PowerPoint slide