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Effect of norcantharidin on proliferation and apoptosis of hepatoma HepG2 cells
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Abstract:
Objective:To study the influence of norcantharidin (NCTD) on the proliferation, apoptosis and survivin expression of human hepatoma HepG2 cells. Methods: MTT assay was used to investigate the effect of NCTD on proliferation of HepG2 cells; Hochest 33258 staining, Annenxin V/PI staining, and agarose gel electrophoresis assay were used to examine the effect of NCTD on apoptosis of HepG2 cells. Western blotting analysis was used to examine the effect of NCTD on survivin expression in HepG2 cells. Results: Proliferation of HepG2 cells was markedly inhibited by NCTD in a dose-dependent manner (5, 10, 20, 40 μg/ml), with the inhibitory rate being (81.27±3.25)% after 40 μg/ml NCTD treatment for 48 h. After HepG2 cells were treated with NCTD for 24 h, fuorescence microscopy revealed that the cell nuclei were condensed and fragmented, and agarose gel electrophoresis of DNA showed a typical DNA ladder map of apoptosis. Flow cytometry results showed that the apoptosis rates of HepG2 cells were (7.33±0.25)%, (18.23±119)%, (32.5±2.30)%, and (48.23±1.17)% after 5, 10, 20, and 40 μg/ml NCTD treatments, respectively. Western blotting analysis results demonstrated that the survivin expression in HepG2 cells was significantly down-regulated with the increase of NCTD dosage. Conclusion: NCTD can inhibit proliferation and induce apoptosis of HepG2 cells, which is related to the down-regulation of survivin protein.
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Project Supported:
Project supported by the Scientific Research Foundation of Health Bureau of Hubei Province（No.JX3B22）
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