ZNF366 Inhibits The Proliferation And Metastasis Of Non-small Cell Lung Carcinoma By Regulating Hippo Pathway And Wnt Pathway Activity

Background:Lung cancer is one of the most common types of malignant cancer in the world and has become the first cause of death from human malignant tumors worldwide.Non-small cell lung carcinoma(NSCLC)accounts for about 85% of all lung cancer types,and because most patients are asymptomatic in the early stages,about 75% of patients are in the middle and advanced stages at the time of discovery,and the 5-year survival rate is very low.ZNF366(also known as DC-SCRIPT)is a zinc finger transcription factor that plays an important role in regulating mammalian dendritic cell differentiation,and studies have shown that ZNF366 expression is an independent marker of good prognosis for breast cancer.However,the expression and mechanism of ZNF366 in lung cancer are unknown,so we hope to explore the function of ZNF366 in NSCLC.Methods : Firstly,the m RNA expression of ZNF366 in LUSC(lung squamous carcinoma,LUSC)and LUAD(lung adenocarcinoma,LUAD)was biologically analyzed through the databases TCGA and GEPIA,and its relationship with the adverse prognosis of lung cancer was evaluated.Subsequently,the expression of ZNF366 in NSCLC tissues and adjacent normal tissues was detected by immunohistochemical staining,and then the experimental results were statistically analyzed to analyze the correlation with the clinicopathological factors of NSCLC.At the same time,the expression of ZNF366 in NSCLC cell lines was detected by Western blot experiment,and the expression of ZNF366 in NSCLC cell lines was detected by cell immunofluorescence experiments.After that,ZNF366 was overexpressed and interfered with in NSCLC cell lines,and the effects of ZNF366 on the invasion and migration ability of NSCLC cells were detected by Transwell experiment,and the effects of ZNF366 on the proliferation function of NSCLC cells were explored by MTS and cloning formation experiments.The Western blot experiment was used to detect the effects of overexpression and interference with ZNF366 on key proteins in the Hippo signaling pathway,classical Wnt signaling pathway and EMT(Epithelial-mesenchymal transition,EMT)process.Finally,the coimmunoprecipitation experiment was used to detect the effect of ZNF366 on the star molecules YAP and SAV1 in the Hippo pathway.Results:1.Using TCGA and GEPIA database analysis,it was found that ZNF366 was poorly expressed in lung cancer tissues.2.Compared with normal lung tissue and human bronchial epithelial cell HEBE,the expression level of ZNF366 in lung cancer tissues and lung cancer cell lines was down-regulated,consistent with the analysis of the database,and the statistical results showed that ZNF366 expression was related to T stage in tumor TNM stage,and low ZNF366 expression was associated with poor prognosis.3.GO and KEGG pathway enrichment analysis showed that ZNF366 was related to the Hippo pathway and the Wnt pathway.4.The results of cell immunofluorescence experiments showed that ZNF366 was mainly localized in the nucleus.5.Overexpression of ZNF366 in A549 cells inhibited the proliferation,invasion and migration ability of NSCLC cells,while the opposite result occurred after the application of si RNA knockdown ZNF366.6.After overexpression of ZNF366 in A549 cells,the expression level of the main protein in the EMT process decreases,and the opposite result occurs after interference.7.The results of co-immunoprecipitation experiments showed that ZNF366 could interact with SAV1 and YAP.8.After overexpression of ZNF366 in A549 cells,the Hippo pathway is activated,the expression of the main component protein LATS1 in the pathway increases,the expression of YAP decreases,and the Wnt pathway is inhibited.After knocking down ZNF366,the expression of LATS1 decreased,the expression of YAP increased,and the Wnt pathway was also activated,resulting in the complete opposite.These data suggest that ZNF366 can regulate the Hippo pathway and the Wnt pathway,affecting the biological function of NSCLC cells.Conclusion : ZNF366 expression was downregulated in non-small cell lung cancer,affecting the cell biological function of non-small cell lung cancer.ZNF366 can inhibit the malignant phenotype of lung cancer cells by modulating the Hippo pathway and the Wnt pathway.