Expression Of EPGN In Head And Neck Squamous Cell Carcinoma And The Mechanism Of EPGN Promoting Invasion And Metastasis Of Nasopharyngeal Carcinoma

Purpose:EPGN(Epithelial mitogen)is one of the ligands of epidermal growth factor receptor(EGFR).Until now,only few researchs were focused on the relationship between EPGN and the development of head and neck squamous cell carcinoma(HNSCC).Especially the relationship between EPGN and the proliferation,invasion and metastasis of nasopharyngeal carcinoma.The aim of our research is to study the role of EPGN in HNSCC,especially in the invasion and metastasis of nasopharyngeal carcinoma.To provide a new molecular markers for the diagnosis and treatment of nasopharyngeal carcinoma.And to furtherly study the possible molecular mechanisms of its biological function.Experimental Design:1、Detection of EPGN expression in different TNM stage of HNSCC tissue microarray HN803b by immunohistochemistry.2、Downloading the GSE39366 data set related to the expression of EPGN in the HNSCC from the GEO database,to study the possible biological function or signal pathway related to the high expression of EPGN in HNSCC through the gene enrichment analysis.3、Construction of the overexpressing and RNAi plasmid of EPGN,then transfected into nasopharyngeal carcinoma CNE-1、CNE-2 cell lines,respectively.4、To detect the influence of EPGN on the proliferation,apoptosis,motility,invasion and adhesion ability of nasopharyngeal carcinoma CNE-1、CNE-2 cell line through CCK-8,Tunel,scratch assay,traswell and angiogenesis assay,respectively.As well as the influence on the angiogenesis of HUVEC cells.5、Detection of EGFR expression in different TNM stage of HNSCC tissue microarray HN803d by immunohistochemistry.6、To confirm the biological function relationship between EPGN and EGFR through EGFR monoclonal antibody and scratch assay.7、To detect the molecular mechanism between EPGN and the motility,invasion of nasopharyngeal carcinoma CNE-1、CNE-2 cell lines through western blot.And to confirm the relationship between EPGN and EGFR furtherly.Result:1、Through immunohistochemical stain for EPGN,EPGN expressed lower in normal head and neck tissue than in HNSCC(P<0.05).But,there is no statistical difference in different T stage and different pathological grade in the HNSCC tissue microarray(P>0.05).2、The result of gene enrichment analysis demonstrate that the over-expression of EPGN is related to the metastasis of HNSCC.3、Silencing EPGN inhibited the proliferation,motility,invasion and adhesion ability of nasopharyngeal carcinoma CNE-1 CNE-2 cell line(P<0.05),but increased the apoptosis ratio conversely(P<0.05).4、Overexpressing EPGN enhanced the angiogenesis ability of HUVEC cells(P<0.05).5、Through immunohistochemical stain for EGFR,EGFR expressed lower in normal head and neck tissue than in HNSCC(P<0.05).But,there is no statistical difference in different TNM stage and different pathological grade in the HNSCC tissue microarray(P>0.05).6、The result of EGFR monoclonal antibody and scratch assay confirmed that EPGN can promote the motility of the nasopharyngeal carcinoma CNE-1、CNE-2 cell line by interact with its receptor EGFR.7、Overexpression of EPGN increased the expression of mesenchymal markers(Vimentin,MMP-9)and decreased the expression of epithelial marker E-cadherin.Conclusion:EPGN expressed more significantly in N2 stage of HNSCC tissue than that in NO stage of HNSCC tissue;Overexpressing EPGN enhanced the angiogenesis ability of HUVEC cells;Silencing EPGN inhibited the proliferation,motility,invasion and adhesion ability of nasopharyngeal carcinoma CNE-1、CNE-2 cell line,but increased the apoptosis ratio conversely;Overexpression of EPGN increased the expression of mesenchymal markers(Vimentin,MMP-9)and decreased the expression of epithelial marker(E-cadherin);EPGN can promote the motility of the nasopharyngeal carcinoma CNE-1、CNE-2 cell line by interact with its receptor EGFR.