Inhibition Of The Hedgehog Signaling Pathway And The Effect On Notch Signaling Pathway In Triple Negative Breast Cancer Cells MDA-MB-231

Objective:Triple negative breast cancer is a type of breast cancer subtype.It is characterized by the non-expression of estrogen receptor(ER),progesterone receptor(PR),and human epidermal growth factor receptor 2(HER-2).Very aggressive,Poor prognosis.Hedgehog and Notch signaling pathways increase in breast cancer.It is closely related to the occurrence,metastasis,and predictive outcomes of triple negative breast cancer.Cyclopamine can reduce the activity of Hedgehog signaling pathway.Significantly reduced breast cancer cell proliferation.Studies have shown that there is an interactive dialogue between the Hedgehog and Notch signaling pathways.Since there is an interactive dialogue between the two,Inhibition of triple negative breast cancer Hedgehog signaling pathway may affect the activity of Notch signaling pathway.To this end,In this paper the influence next target face-MB-231 cells of MDA Hedgehog signaling pathway,to investigate the expression of Smo and Gli-1 in the Hedgehog signaling pathway,And whether it affects the expression of Notch-1 and Hes-1,key molecules of the Notch signaling pathway.Methods:The MDA-MB-231 cells purchased from the Chinese Academy of Sciences Cell Bank were cultured.According to the dosing concentration,the experiments were divided into 4 groups:Cy 0 group(targeting concentration in the ring is 0 μmol/L group),Cy 10 group(targeting concentration in the ring is 10 μmol/L group),Cy 20 group(targeting concentration in the ring is 20 μmol/L group),Cy 40 group(targeting concentration in the ring is 40 μmol/L group),After 48 hours of ring target treatment,mRNA and protein were extracted.The expression levels of Smo,Gli-1,Notch-1 and Hes-1 mRNA in cells were detected by RT-PCR.Smo,Gli-1,1 Notch-Hes-1 expression and protein was detected by Western blot,The final statistical analysis.Results:(1)Cyclopamine respectively 0,10,20,40 μmol/L concentration the role of cell 48h,The expression of Smo protein was detected by Western blot.The expression of mRNA was detected by real-time quantitative PCR.Turn out,After different concentrations of ring target treatment of MDA-MB-231 cells,Smo protein expression decreased,The higher the drug concentration,the greater the decrease in protein expression(P<0.01).After different concentrations of ring target treatment of MDA-MB-231 cells,Smo mRNA expression decreased,The higher the drug concentration,the greater the decrease in mRNA expression(P<0.01).Same,After different concentrations of ring target treatment of MDA-MB-231 cells,Gli-1 protein expression decreased The higher the drug concentration,the greater the decrease in protein expression(P<0.01).After different concentrations of ring target treatment of MDA-MB-231 cells,Gli-1 mRNA expression decreased,The higher the drug concentration,the greater the decrease in mRNA expression(P<0.01).(2)Cyclopamine respectively 0,10,20,40 μmol/L concentration the role of cell 48h,The expression of mRNA was detected by Western blot and real-time quantitative PCR.Turn out:After different concentrations of ring target treatment of MDA-MB-231 cells,There was no significant change in the expression of Notch-1 protein and it was not related to the concentration of the drug(P>0.05).After different concentrations of ring target treatment of MDA-MB-231 cells,There was no significant change in the expression of Notch-1 mRNA,and it was not related to the concentration of the drug(P>0.05).Same,After different concentrations of ring target treatment of MDA-MB-231 cells,There was no significant change in the expression of Hes-1 protein and it was not related to the concentration of Hes-1 protein(P>0.05).After different concentrations of ring target treatment of MDA-MB-231 cells,The expression of Hes-1 mRNA had no significant changes,and it had no relation with the concentration of Hes-1(P>0.05).Conclusion:(1)Cyclopamine specifically inhibit the Hedgehog signaling pathway,By treatment in vitro MDA-MB-231 cells,Different concentrations of Cyclopamine can inhibit the expression of key molecules Smo and Gli-1 in Hedgehog signaling pathway.And as the concentration increases,the inhibitory effect gradually increases.(2)Cyclopamine acts on MDA-MB-231 cells in vitro,Does not affect the expression of Notch-1 and Hes-1 key molecules in the Notch signaling pathway.(3)Inhibition of Hedgehog signaling pathway in TNBC does not affect the activity of Notch signaling pathway.