The Activation Of Notch-1Signal That Effect For The Epithelial-mesenchymal Transformation And The Biology In The Esophageal Squamous Cell Carcinoma

Objective:To study the regulational function of Notch-1signaling pathway for epithelial-mesenchymal transformation in the Eca-109.Clear the change of proliferation,apoptosis and migration ability in the Eca-109after the Notch-1signaling pathwayactivated.Methods:Know the mechanisms of Notch-1signaling pathways activation for theepithelial-mesenchymal transformation in the esophageal squamous cell carcinoma,The experiment was divided into the group of Control, DMSO, MOCK, N1ICD,TGF-β1, DAPT+TGF-β1, DAPT.LV-N1ICD transfected the Eca-109toup-regulation Notch-1signaling pathway;After72hours, according to the groupingsituation respectively adding5ng/ml TGF-β1induced the esophageal squamous cellcarcinoma(Eca-109) occurred EMT and15μmol/ml DAPT inhibition the Eca-109occurred EMT.After48hours,detected the cell viability by CCK-8; Western blottingdetected the protein expression change of E-cadhenrin,Vimentin,Notch-1,Hes-1,Hey-1,Jagged-1.after5W,Crystal violet stained and counted the clones.observed themigration ability changed of Eca-109during24hours. AnnexinV/PI dyed anddetected cell apoptosis.Result:1The CCK8test cell vitality results showed that the proliferation vitality of theEca-109was the strongest in the group of N1ICD, compared with the group ofControl, MOCK, TGF-β1，TGF-β1+DAPT, DMSO, DAPT was significant difference（P<0.05）.In the group of DAPT, the Eca-109proliferationability was the weakest,compared with other group was significant difference.Compared with the group ofDMSO, there was no significant difference.2The cell clone count results showed that the group of N1ICD compared withthe group of Control、MOCK、TGF-β1、DAPT+TGF-β1和DMSO、DAPT，Eca-109clone formation rate and the number increased obviously, there were statistically significant differences（P<0.05）.3The scratch test results showed that the group of N1ICD compared with thegroup of Control、MOCK、TGF-β1、DAPT+TGF-β1和DMSO、DAPT，The closureof scratch wound of Eca-109was facilitated obviously, there were statisticallysignificant difference (P <0.05).4Flow cytometry instrument detected the apoptosis of Eca-109results showedthat the group of N1ICD compared with the group of Control、MOCK、TGF-β1、DAPT+TGF-β1和DMSO、DAPT，the percentage of apoptotic cells decreasedsignificantly,there were statistically significant difference (P <0.05).5Western-blotting test results showed that the protein expression of E-Cadherinwas significantly reduction in the group of N1ICD,compared with the group ofControl,MOCK,TGF-β1,DAPT+TGF-β1和DMSO,DAPT,there was a significantdifference (P <0.05).the proteins expression of Vimentin,Notch-1, Hes-1,Hey-1、Jagged-1were significantly increment in the N1ICD+TGF-β1group,compared withthe group of Control,MOCK,TGF-β1,DAPT+TGF-β1和DMSO,DAPT,there was asignificant difference (P <0.05).Conclusion:The Lentiviral Vector-N1ICD(LV-N1ICD) transfected Eca-109to up-regulationthe Notch1signaling pathways,It obviously promote the epithelial-mesenchymal(EMT) transformation in the esophageal squamous cell carcinoma(Eca-109),and As the activation of the Notch-1signal,the proliferation, migration andapoptosis resistance ability of Eca-109were significantly enhanced.