| Objectives: To detect the proliferation inhibition on human hepatoma cells and expression of β-catenin, C-myc and Cyclin D1 genes/proteins in Wnt pathway induced by Esculetin, and explore its anti-tumor mechanism.Method:1. 3-(4,5-dimethyithiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) test was used to determine the Inhibition ratio of Esculetin on human hepatoma cells SMMC-7721 and human nomal liver cells HL-7702. Visualization of cytopathologic effect (CPE) assay was used to deteimine the inhibition effect of Esculetin on human hepatoma cells SMMC-7721.2. Real-time PCR was used to quantify the expression of P-catenin,C-myc and Cyclin D1 genes, at different time points and concentrations, respectively.3. Western blot was used to detect the expression of P-β-catenin, β-catenin,C-myc and Cyclin D1 of different treatment groups.Result:1. Esculetin inhibited the proliferation of human hepatoma cells SMMC-7721 in a time- and dose-dependent manner. Esculetin had less cytotoxicity on human nomal liver cells HL-7702 than Cisplatin at the same inhibiton ratio on human hepatoma cells SMMC-7721.2. Real-time PCR was conducted to analyse β-catenin,C-myc and Cyclin D1 genes at different time points. Compared with cell control, Esculetin group down-ragulated the expression of β-catenin, C-myc and Cyclin D1 at 48 and 72h(P<0.05).3. Western blot detection was peformed to analyse P-β-catenin, β-catenin,C-myc and Cyclin D1 proteins. Esculetin increased the expression of P-β-catenin at 24h. The expression of β-catenin, C-myc and Cyclin D1 were suppressed after treatment with Esculetin for 72h.Conclusion:1. Esculetin inhibited the proliferation of human hepatoma cells SMMC-7721 in a time-and dose-dependent manner.2. Esculetin enhanced P-β-catenin expression and promoted the degradation of β-catenin,then inhibited the activation of Wnt signaling pathway.3. Esculetin inhibited the proliferation of human hepatoma cells SMMC-7721 by down-regulating the expression of β-catenin,C-myc and Cyclin D1. |
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