| Objective:To investigate the effects of apigenin on the proliferation,migration and invasion of Tca8113 cells,and explore its possible molecular mechanisms,and provide new experimental basis for clinical antitumor effect of apigenin.Method:Tca8113 cells were treated with apigenin.The proliferation ability were investigated by MTT assay.The migration,ability were investigated by wound-healing test.The invasion ability were investigated by Transwell chamber assay.The intracellular localization of total-MEKKl,p-MEKKl and p-ERK1/2 in Tca8113 cells was detected by immunofluorescence staining.The expression level of MEKK1,ERK1/2,E-Cadherin and Vimentin were detected by Western Blot.Results:(1)The proliferation of Tca8113 cells were significantly inhibited by treatment of 50,75 and 100 μmol/L of apigenin for 24,48 and 72h compared with control group(P<0.01),and inhibited by treatment of 75 and 100 μmol/L of apigenin for 24,48 and 72h compared with 50 μmol/L of apigenin group were significantly(P<0.01 or P<0.05),also inhibited by treatment of 100 pmol/L of apigenin for 72h compared with 75 μmol/L of apigenin group were significantly(P<0.01),which showed dose dependent manner.Interestingly,the proliferation of Tca8113 cells were significantly inhibited by treatment of 50 μmol/L of apigenin at 48 and 72h compared with 24h group(P<0.01),and inhibited by treatment of 75 μmol/L of apigenin at 72h compared with 24h group were significantly(P<0.05),also inhibited by treatment of 100 p.mol/L of apigenin at 72h compared with 24h and 48h group were significantly(P<0.01),which showed time dependent manner.(2)Also,the repairing rate of Tca8113 cells were significantly decreased than the control group after treated with 50,75 and 100 μmol/L of apigenin for 24,48 and 72 h(P<0.01),which showed time and dose dependent manner.(3)The number of Tca8113 cells that invaded the underside of the porous polycarbonate membrane were significantly decreased than the control group after treated with 50,75 and 100 μmol/L of apigenin in the below chamber for 48h,and the relative invasion rate were 75%,45%and 30%respectively,which showed dose dependent manner.(4)In Tca8113 cells the total-MEKK1 mainly located in the cytoplasm,the p-MEKK1 mainly located in the cytoplasm and nuclear envelope;As the increasing of drug concentration,the fluorescence density located in the cytoplasm was decreased than the control group.The p-ERK1/2 mainly located in the cytoplasm and nuclear;As the increasing of drug concentration,the fluorescence density located in the cytoplasm and nuclear was decreased than the control group.(5)Following treatment with apigenin at 50,75 and 100 pmol/L for 48h,the p-MEKK1 protein level was decreased than the control group(P<0.01),and the p-ERKl/2 protein level was decreased after treated with 75 and 100 μmol/L of apigenin for 48h(P<0.01),which showed dose dependent manner,whereas the total-MEKKl and total-ERK1/2 protein levels were unchanged in Tca8113 cells detected by Western Blot.(6)Following treatment with apigenin at 50,75 and 100 μmol/L for 48h,the E-Cadherin protein levels were increased,whereas Vimentin were decreased than the control group in Tca8113 cells detected by Western Blot(P<0.01),which showed dose dependent manner.Conclusion:(1)Apigenin significantly inhibit the proliferation,horizontal migration and invasion of Tca8113 cells.(2)The anti-tumor mechanism of apigenin may be associated with MAPK signaling pathways.(3)The anti-tumor mechanism of apigenin may be associated with Epithelial-mesenchymal transition. |
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