The Transcripotomes Of Interactions Between Nasopharyngeal Carcinoma And Lymphatic Endothelial Cell

Objective:To establish a lymphatic endothelial cell microenvironment of nasopharyngeal carcinoma under co-culture of nasopharyngeal carcinoma and lymphatic endothelial cell.To explore the interactions between co-cultured nasopharyngeal carcinoma and lymphatic endothelial cell by RNA-Seq detecting on differentially expressed genes.To find the nasopharyngeal lymphatic metastasis associated genes for further clarifying the mechanisms of nasopharyngeal carcinoma lymph node metastasis.Methods:Red fluorescence-labeled lentivirus and green fluorescence-labeled lentivirus were respectively used to transfect nasopharyngeal carcinoma line 5-8F cells and lymphatic endothelial cell(LEC),and flow cytometry was used to detect the transfection efficiency.Morphological changes of transfected 5-8F cells and LEC were observed with live cell imaging system.After co-cultured for 72h,cells were sorted with flow cytometry.RNA-Seq was used to detect differentially expressed genes of 5-8F cells and LEC before and after co-culture,and the result was analyzed with bioinformatics.The expression profile data of 3 selected gene were analyzed with Realtime-PCR for verification.Results:1.The observation of live cell imaging system showed that LEC grow around 5-8F cells under co-culture.The athletic ability of LEC was stronger than 5-8F cells and the athletic ability of some 5-8F cells was enhanced when contacting with LEC.2.Significant difference was observed when analyzing the expression profile data of differentially expressed genes in co-cultured tumor cells and lymphatic endothelial cells.3.Differentially expressed genes with significantly enriched GO function were mainly involved in cell adhesion,cell proliferation,transcription regulation,signal transduction and so on.4.The function enrichment Pathway of differentially expressed genes of 5-8F cells and LEC was 9 and 8 respectively.5.Realtime-PCR showed that the overall trend of differentially expressed multiple relationship and sequencing results was consistent.Conclusion:1.Significant differences were found in expressed genes of 5-8F cells and LEC under co-culture,and interactions was observed between 5-8F cells and LEC.2.The analysis of function enrichment Pathway with KEGG database on differentially expressed genes suggested that lymphatic endothelial cell microenvironment may prompt epithelial-mesenchymal transition of nasopharyngeal carcinoma through MAPK signal pathway.Lymphatic endothelial cell may prompt proliferation and induce epithelial-mesenchymal transition of nasopharyngeal carcinoma through important signaling pathways of cancer.