Study On The Mechanism Of Notch Signaling Pathway Inhibitor DAPT Inhibiting The Invasion And Migration Of Tongue Squamous Carcinoma Cell By Regulating LncRNA-KAT14

Research background:The treatment of advanced tongue squamous cell carcinoma has encountered a bottleneck due to its poor efficacy and low survival rate.Studies have shown that there is diversity in the Notch signaling pathway.According to the available data,it is clear that Notch has the function of bidirectional regulation,and further studies in tongue squamous cell carcinoma are lacking.LncRNAs are currently believed to have an influence on the physiological functions of cells.According to the results of previous studies,a variety of lncRNAs are changed in tongue squamous cell carcinoma cells,which can be up-regulated or down-regulated.Our research group found that lncRNA increased more than twice in more than 400 sites.Therefore,we speculate that lncRNA-KAT14 plays a role in oral squamous cell carcinoma.Objective:The tongue squamous cell carcinoma cells CAL-27 and SCC-9 were cultured,and DAPT culture was added to observe their effects on the proliferation,invasion and migration of tongue cancer cells.The proliferation,invasion and migration of tongue cancer cells were detected by CCK-8 method,cell scratch test,plate clone formation test,and cell migration and invasion test.The expressions of epithelial mesostromal associated proteins N-cadherin,E-cadherin,Vimentin and Snail were detected by Western blot assay,and the changes of lncRNA-KAT14 in tongue cancer were detected by real-time PCR assay.Further,by silencing lncRNA-KAT14,the effects of lncRNA-KAT14 on the proliferation,invasion and migration of cancer cells and the changes of related proteins were detected in the same method.LncRNA profiles of tongue cancer cells were detected by high throughput RNA sequencing.Methods:The tongue squamous cell carcinoma cells CAL-27 and SCC-9 were cultured,and DAPT culture was added to observe their effects on the proliferation,invasion and migration of tongue squamous cell carcinoma cells.The proliferation,invasion and migration of tongue squamous cell carcinoma cells were detected by CCK-8 method,cell scratch test,plate clone formation test,and cell migration and invasion test.Western blot assay was used to detect the expressions of epithelial mesostromal associated proteins N-cadherin,E-cadherin,Vimentin and Snail.Real-time PCR assay was used to detect the changes of lncRNA-KAT14 in tongue squamous cell carcinoma.Further,by silencing lncRNA-KAT14,the effects of lncRNA-KAT14 on the proliferation,invasion and migration of cancer cells and the changes of related proteins were detected in the same method.LncRNA profiles of tongue squamous cell carcinoma cells were detected by high throughput RNA sequencing.Results:DAPT can reduce the proliferation,invasion and migration of tongue squamous cell carcinoma cells.The expression of lncRNA-KAT14 is decreased in tongue squamous cell carcinoma,and the addition of DAPT can overexpress lncRNA-KAT14 in tongue squamous cell carcinoma,and alter the expression of epithelial mesenchymal transformation related proteins,thus inhibiting the proliferation,invasion and migration ability of cancer cells.When lncRNA-KAT14 was silenced,it was found that the invasion ability of tongue cancer cells was increased.When DAPT was added,the proliferation,invasion and migration ability of tongue cancer cells were also decreased,and the expression of lncRNA-KAT14 was increased,that is,the overexpression of lncRNA-KAT14 was induced by DAPT.RNA-sequencing samples predicted an increase in lncRNAs in tongue cancer cells after DAPT treatment,including 1919 new transcripts in the 200?M group and 2668 in the 500?M group,while only 269 in the control group.Our results showed that KAT14(Gene ID 57325)was significantly higher in the two DAPT intervention groups(6,00825 and 8.1327,respectively),while there was little KAT14 in the non-intervention group.Conclusion:DAPT can inhibit the proliferation,invasion and migration of tongue squamous cell carcinoma cells,and can obviously induce the overexpression of lncRNA-KAT14.DAPT may inhibit the proliferation,invasion and migration of tongue squamous cell carcinoma cells by regulating the epithelial-mesoplasm-related proteins N-cadherin,E-cadherin,Vimentin and Snail.The expression of KAT14 increased after DAPT treatment,suggesting that DAPT may induce the overexpression of lncRNA-KAT14 and affect the proliferation,invasion and migration of tongue cancer cells.