Effects On Cell Proliferation And NF-κB Pathway In Neuroblastoma After Inhibition Of Notch Pathway By DAPT

Objectives:1,To evaluate the effect ofγ-secretase inhibitor DAFT on cell proliferation of human neuroblastoma SH-SY5Y.2,To observe the effect of DAPT on Notch signaling molecules,including Notch1 and Hes1.3,To study the interaction between Notch and NF-κB pathways and its potential mechanisms in neuroblastoma SH-SY5Y after DAPT treatment.Methods:MTT assay to find the best drug concentration and the best treatment time after different concentrations of DAPT treatment in SH-SY5Y cells;as the role of drug concentration and time to deal with cells,we use RT - PCR,Western blotting to detect expression changes of some cell proliferation associated geneResults:1,DAPT inhibited SH-SY5Y cell growth effectively in a time- and dose-dependent manner.2,5μM DAPT can block Notch signaling pathway.After 48 hours' treatment, DAPT down-regulated expression of NICD and Hesl,and then also down-regulated expression of p65 with its target genes Cyclin D1,Bcl-2.Conclusions:1,DAPT effectively blocked Notch signaling transduction and inhibited cell growth in SH-SY5Y.2,In neuroblastoma SH-SY5Y,following block of Notch signaling,p65 expression is down-regulated,and a similar effect on Cyclin D1,Bcl-2. These mean a cooperation relationship of Notch and NF-κB pathways...