The M+echanism Of Epstein-Barr Virus Encoded LMP1 Protein Affects The Proliferation Of NK/T Cell Lymphoma Through Glycolytic Pathway

Background and PurposeNK/T-cell Lymphoma(NKTCL)was mainly found in East Asia and Latin America,and was relatively rare in European and American countries.Epstein-Barr virus infection plays an important role in the occurrence and development of NKTCL.At the present,the pathogenesis and effective treatment of NKTCL are still under study.Immunotherapy has made good progress in the treatment of NKTCL in recent years,but the selection of immune checkpoints is still worthy of further investigation.Cell metabolism plays a very important role in the development of tumor.Therefore,this study focused on the role of the oncogenic protein latent membrane protein 1(LMP1)encoded by Epstein-Barr virus on the glycolytic metabolism of NKTCL and its mechanism.Materials and Methods1 Metabolomics was used to screen and identify the metabolites and metabolic pathways of Epstein-Barr virus positive NK lymphoma cells(LMP1+)(NKL,NK92),Epstein-Barr virus negative NK lymphoma cells(LMP1-)(YTS,Kai3,SNK6)and normal NK cells.2 The effect of glycolytic activity on cell proliferation,apoptosis and drug sensitivity of Epstein-Barr virus positive NK lymphoma cells(LMP1+)(NKL,NK92).The CCK-8 experiment was used to determine the cell OD values of NKL and NK92 with glycolysis inhibitors(experimental group)and without glycolysis inhibitors only added with DMSO(control group).The OD values were measured at 24 hours,48 hours,and 72 hours.The change in the graph depicts the proliferation curve of the cell.Flow cytometry was used to determine the apoptosis ratio of the experimental group(with glycolysis inhibitor added)and the control group.The CCK-8 experiment was used to determine the cell OD value of each group and calculate the gemcitabine(Gem)IC50 value to compare the drug sensitivity of each group.3 The candidate protein molecules that interact with LMP1 were identified by co-immunoprecipitation combined with protein profilometry,and the interacting protein molecules that have potential influence on signaling pathway activity were screened.Result1 The study suggested that the glycolytic level of Epstein-Barr virus positive lymphoma cells(NKL,NK92)was significantly higher than that of Epstein-Barr virus negative lymphoma cells(YTS,KAI3,SNK6)and normal NK cells.The glucose uptake rate and lactic acid production rate were higher than those of Epstein-Barr virus negative lymphoma cells and normal NK cells,and Epstein-Barr virus positive NK/T cell lymphoma cell line gemcitabine(GEM)had lower drug sensitivity than Epstein-Barr virus negative NK/T cell lymphoma cell line.2 After adding glycolysis inhibitors to the medium of EB virus-positive NK lymphoma cell lines(YT,SNK6),the proliferation ability of tumor cells was significantly weakened(P<0.001),and the proportion of tumor cell apoptosis increased(P<0.001),Tumor cells are more sensitive to drugs(P<0.001).3 Protein molecules interacting with LMP1 in Epstein-Barr virus positive tumor cells(LMP1+)were identified by co-immunoprecipitation combined with protein profilometry,including key protein molecules such as TRAF3 and RPL4 that may have important effects on downstream signaling pathways.Conclusion1 The glycolytic activity of Epstein-Barr virus positive NK/T lymphoma cells was higher than that of Epstein-Barr virus negative NK/T lymphoma cells.2 After inhibiting glycolytic activity,the proliferation ability of NK/T lymphoma cells decreased,apoptosis increased,and drug sensitivity increased.3 The LMP1 protein encoded by Epstein-Barr virus may affect the downstream signaling pathway through binding with TRAF3 and other molecules,and then regulate the glycolysis of tumor cells.