LMP1-mediated glycolysis induces myeloid-derived suppressor cell expansion in nasopharyngeal carcinoma
Fig 2
LMP1 increased glycolysis and the expression of glycolysis-related genes in NPC cells.
(A) Exogenous EBV oncoprotein LMP1 was stably overexpressed in the NPC cell lines CNE2 and TW03 following lenti-LMP1 transfection, and the lenti-vector was included as control. WB showed that the expression of LMP1 and GLUT1 was increased in CNE2-LMP1 and TW03-LMP1 cells compared with that in CNE2-vector and TW03-vector cells, and GAPDH was included as a control. One representative experiment of five independent experiments is shown. (B-D) ECAR assay results. CNE2-vector, CNE2-LMP1, TW03-vector and TW03-LMP1 cells were cultured in base DMEM media with no glucose or glutamine. ECAR was assessed after the addition of 10 mM glucose and in response to the metabolic inhibitor oligomycin and the glucose inhibitor 2-DG. The time course and calculations for (B) glycolytic capacity and (C) statistical analysis of glycolytic capacity and glycolytic reserve are shown. (D) The relative mRNA levels of glycolysis-related genes, including GLUT1, HK2, GPI, PFKB1, 2, 3 and 4, ALDOA, PGK1, PKM2 and LDHA, in CNE2-vector, CNE2-LMP1, TW03-vector and TW03-LMP1 cells were measured via real-time qRT-PCR. Statistical results are representative of at least three experiments performed in triplicate. All values are shown as the means ± SEM; * indicates P < 0.05. ** indicates P < 0.01.