| Objective:To investigate the effect of Berberine on the apoptosis and angiogenesis whether associated with GRP78 expression and Wnt/β-catenin signaling pathway of SW480 respectively.Methods:MTT assay and Wound healing assay were used to examine cell proliferation and migratory capacities of SW480 cells respectively.Apoptosis was detected by using flow cytometry method.The capabilities of angiogenesis of SW480 cells were assessed by 3D cultures and Chick Chorioallantoic Membrane(CAM)assay.Immunofluorescence was used to estimate the nucleuslocalization of β-catenin.The changes in expression of GRP78,Bcl-2,Bax,GRP94,β-catenin,c-Myc,cyclin D1,Vimentin,cytokeratin,VEGF and HIF-1α proteins were detected by Western blot.Results:1.Compared with the control,the proliferation of SW480 and HT-29 cells was significantly inhibited by different concentrations of Berberine.The proportion of apoptotic cell numbers drastically was increased in a dose-dependent manner at the concentration range from 20 to 100 μmol/L.Accordingly,the protein level of GRP78 was down-regulated following 50 μmol/L treatment.Additionally,the expression of Bax was induced by Berberine,while Bcl-2 was decreased.The expression of all the proteins was reversed when GRP78 was stimulated by BiX.2.Compared with the control,the migratory capacities and the capabilities of angiogenesis of SW480 cells were inhibited significantly by 50 μmol/L of Berberine.β-catenin expression,as well as its nucleus localization were inhibited following 50μmol/L of Berberine.Moreover,the expression of c-Myc,cyclin D1,Vimentin,VEGF and HIF-laare was decreased by Berberine,while cytokeratinis were increased.But the expression of all of the protein was reversed when Wnt/β-catenin signaling pathway was actived by LiCl.Conclusion:1.Berberine can induce the apoptosis of SW480 cells by down-regulating the expression of GRP78.2.Bererine can inhibit angiogenesis of SW480 cells by down-regulating Wnt/β-catenin signaling pathway. |
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