摘要
Objective： To investigate the effect ofberbamine on human hepatoma cell line SMMC7721. Methods： The effects of 24 h and 48 h incubation with different concentrations （0-64 μg/ml） of the berbamine on SMMC7721 cells were evaluated using 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide （MTT） assay. Hoechst 33258 staining was conducted to distinguish the apoptotic cell, and the appearance of sub-G1 stage was determined by PI （propidium iodide） staining, the percentage of apoptotic cell was determined by flow cytometry following annexin V/PI staining. Flow cytometry was performed to analyze the cell cycle distribution and the mitochondrial membrane potential （△ψm）, the expression of activated caspase3 and caspase9 was analyzed by Western-blot. Results： The proliferation of SMMC7721 was decreased after treatment with berbamine in a dose- and time-dependent manner. Berbamine could induce apoptosis in SMMC7721 cells and could cause cell cycle arrest in G0/G1 phase, to induce loss of mitochondrial membrane potential （AVm） and activate caspase3 and caspase9. Berbamine-induced apoptosis could be blocked by the broad caspase inhibitor z-VAD-fmk. Conclusion： Berbamine exerts antiproliferative effects on human hepatocellular carcinoma SMMC7721 cells. The anticancer activity of berbamine could be attributed partly to its inhibition of cell proliferation and induction of apoptosis in cancer cells through loss in mitochondrial transmembrane potential and caspase activation.
Objective:To investigate the effect of berbamine on human hepatoma cell line SMMC7721.Methods:The effects of 24 h and 48 h incubation with different concentrations(0～64 μg/ml) of the berbamine on SMMC7721 cells were evaluated using 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide(MTT) assay.Hoechst 33258 staining was conducted to distinguish the apoptotic cell,and the appearance of sub-G1 stage was determined by PI(propidium iodide) staining,the percentage of apoptotic cell was determined by flow cytometry following annexin V/PI staining.Flow cytometry was performed to analyze the cell cycle distribution and the mitochondrial membrane potential(?ψm);the expression of activated caspase3 and caspase9 was analyzed by Western-blot.Results:The proliferation of SMMC7721 was decreased after treatment with berbamine in a dose-and time-dependent manner.Berbamine could induce apoptosis in SMMC7721 cells and could cause cell cycle arrest in G0/G1 phase,to induce loss of mitochondrial membrane potential(?ψm) and activate caspase3 and caspase9.Berbamine-induced apoptosis could be blocked by the broad caspase inhibitor z-VAD-fmk.Conclusion:Berbamine exerts antiproliferative effects on human hepatocellular carcinoma SMMC7721 cells.The anticancer activity of berbamine could be attributed partly to its inhibition of cell proliferation and induction of apoptosis in cancer cells through loss in mitochondrial transmembrane potential and caspase acti-vation.
基金
Project supported by the National Natural Science Foundation of China (No. 30400521)
the Science and Technology Department of Zhejiang Province (Nos. 2004D31026 and 2002D3007)
the Education Department of Zhejiang Province (No. 20060427), China
