Effects of daphnetin on the proliferation,apoptosis and chemosensitivity of breast cancer by regulating the Shh/Gli1 signaling pathway
Objective To investigate the effects of daphnetin on the proliferation,apoptosis and chemosensitivity of breast cancer by regulating the sonic hedgehog(Shh)/glioma-associated oncogene homologue 1(GLI1)signaling pathway.Methods The human breast cancer cell line MDA-MB-231 was divided into control group(blank control),daphnetin group(40μmol/L daphnetin),glioma-associated oncogene(Gli)antagonist-61(GANT61)group(10μmol/L GANT61),and daphnetin+GANT61 group(40μmol/L daphnetin and 10μmol/L GANT61).Cell proliferation,and migration were assessed by CCK-8 assay and wound healing assay,respectively.After adding 1mg/L doxorubicin(Dox)in above cells for 48 hours,chemotherapy sensitivity and apoptosis were assessed by CCK-8 assay and flow cytometry,respectively.Protein expressions of key proteins in the Shh/Gli1 signaling pathway and apoptosis proteins were detected by Western blot.Results Compared with that of control group,the survival rate of MDA-MB-231 cells significantly decreased with the increasing concentration of daphnetin(P<0.05).There was no significant difference in the survival rate of MDA-MB-231 cells treated with 40μmol/L and 50μmol/L daphnetin(P>0.05).Therefore,40μmol/L daphnetin was chosen as the intervention condition for subsequent experiments.Compared with those of the control group,cells in the daphnetin group and GANT61 group presented significantly lower optical density at 450nm(OD450)value,scratch healing rate,and protein expressions of Gli1 and B cell lymphoma-2(Bcl-2)(P<0.05),and significantly higher protein expression of Bax,apoptotic rate,and chemotherapy sensitivity to Dox(P<0.05).Compared with those of the daphnetin group,cells in the daphnetin + GANT61 group presented significantly lower OD450 value,scratch healing rate,and protein expressions of Gli1 and Bcl-2(P<0.05),and significantly higher protein expression of Bax,apoptotic rate,and chemotherapy sensitivity to Dox(P<0.05).Conclusion Daphnetin may inhibit the proliferation of breast cancer cells,induce apoptosis and improve chemosensitivity by inhibiting the Shh/Gli1 signaling pathway.
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