Effect Of Notch-1 Silencing On Epithelial-mesenchymal Transformation With Bladder Cancer In Vitro

Objective:To explore the effect of Notch-1 silencing on epithelial-mesenchyme transition(EMT) and its molecular mechanism. Methods:Bladder cancer cell line 5637 was transfected with Notch-1 shRNA by using liposome lipofectamine 2000.Then the 5637 cells growth,change and transfection efficiency were observed with inverted microscope. Transfection efficiency was also tested by qRT-PCR and Western-blot. mRNA and protein expression of E-cadherin、N-cadherin and vimentin were detected by qRT-PCR and western blot respectively.Flow cytometry was used to detect cell apoptosis rate. Transwell method was used to assesse the changes of 5637 cancer cell invasion ability after transfection. Results:The cell number of test group decreased significantly with obvious change in cellular morphology including cell shrinking and irregular shape.Transfection efficiency was 80%. qRT-PCR and western-blot results showed that Notch-1 was down-regulated after transfection(p<0.05).Furthermore,vimentin was significantly down-regulated and E-cadherin、N-cadherin actin was significantly up-regulated after transfection(p<0.05)The apoptosis rate of experimental group on early and late stage rised significantly detected by flow cytometry(p<0.05)and invasion ability prominently lowered(p<0.05). Conclusion:Blocking Notch signal pathway by down-regulating Notch-1, EMT of bladder cancer was interdicted partly.Meanwhile,the ability of apoptosis resistance and invasion of bladder cancer cell was declined.