| SonicHedgehog(Shh)and Wnt signaling pathways were found to be activated in moderately differentiated gastric cancer cell SGC-7901,and the role of nuclear transcription factors Gli1 and ?-catenin as Shh and Wnt signaling pathways on SGC-7901 cells remains unclear.Whether curcumin,a Chinese herb,can exert anti-tumor by inhibiting Gli1 and ?-catenin was the focus in our study.Objective:To investigate whether curcumin can affect the migration,invasion,cytoskeleton remodeling,epithelial mesenchymal transformation(EMT),apoptosis and cell cycle of gastric cancer cell line SGC-7901 by inhibiting Gli1 and ?-catenin;whether there was the interaction between Gli1 and ?-catenin in SGC-7901 cells and the effect of curcumin on this interaction.Methods:1.CCK-8 assays were used to observe the ability of curcumin to inhibit the proliferation of SGC-7901 cells to determine the experimental concentration of curcumin.2.Small interference RNA(siRNA)was used to transfect SGC-7901 cells,and Western blotting and qPCR were used to verify whether the siRNA could successfully knock down Gli1 or ?-catenin in SGC-7901 cells at protein level and mRNA level,respectively,thereby achieving inhibition of Shh and Wnt signaling pathways.3.Transwell assays,Indirect immunofluorescence(IF)and flow cytometry(FCM)were used to verify the effects of knock down Gli1 or ?-catenin and curcumin stimulation on the migration and invasion ability,cytoskeleton changes,cell apoptosis and cell cycle of SGC-7901 cells.4.Western blotting was used to detect the changes of Vimentin and E-cadherin related in cell epithelial mesenchymal transition after knockdown Gli1 of Shh signaling pathway or ?-catenin of Wnt signaling pathway and curcumin stimulation in SGC-7901.5.Co-immunoprecipitation assay(Co-IP),Western blotting and qPCR were used to prove whether Gli1 and ?-catenin were correlated in SGC-7901 cells,whether the interaction could be inhibited by curcumin.Results1.Inhibiting the Shh and Wnt signaling pathways of SGC-7901,respectively,reduced the migration and invasion ability of SGC-7901 cells and the cytoskeleton remodeling;promoted the cell cycle to be blocked in G0/G1 and S phases.2.There was an interaction between Gli1 and ?-catenin in SGC-7901 cells,and this interaction can be inhibited by curcumin.3.Curcumin inhibited the migration and invasion ability and EMT process of SGC-7901 cells,caused cytoskeleton remodeling,and promoted apoptosis and cycle arrest by regulating Gli1 and ?-catenin.Conclusion:This study proved that curcumin inhibited the migration,invasion,cytoskeleton remodeling and EMT process of gastric cancer cell SGC-7901,and promoted cell apoptosis and cycle arrest through Gli1 and ?-catenin.It provided a new idea for further study on the pathogenesis of gastric cancer at the molecular level and clinical precise treatment. |
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