[EBNA1 Knockdown Modulates Proliferation, Apoptosis, and Metabolism in Burkitt's Lymphoma Cells by Downregulating miR-127-5p].

OBJECTIVE

To preliminarily elucidates the role of EBNA1 in epigenetically regulating miR-127-5p and influencing the proliferation, apoptosis, and metabolism of Burkitt's lymphoma (BL) cells.

METHODS

The expression of EBNA1 was knocked down in EBV-positive Raji BL cells through lentiviral shRNA transduction. Gene expression, promoter methylation, and apoptosis were evaluated by quantitative real-time PCR, pyrosequencing, and flow cytometry, respectively. Cell proliferation was assessed by the CCK-8 assay, while cellular glycolytic activity was analyzed using the Seahorse XF Glycolysis Rate Assay. A comparative metabolomic study between EBV-positive Raji cells and EBV-negative CA46 BL cells was conducted using UHPLC-QE-MS.

RESULTS

Compared with CA46 cells, the transcriptional level of miR-127-5p was significantly increased in Raji cells (P<0.05). Silencing EBNA1 significantly inhibited Raji cell proliferation, glycolysis level, and miR-127-5p transcriptional level, while promoting Raji cell apoptosis, and increasing Caspase-3 and LCLAT1 protein expression (all P <0.05). Pyrosequencing confirmed that the downregulation of miR-127-5p transcriptional level was associated with increased promoter methylation in EBNA1-shRNAi-Raji cells. Metabolomic analysis revealed that Raji cells produced more tumor metabolism-related metabolites compared to CA46 cells.

CONCLUSION

EBNA1 upregulates the expression of miR-127-5p through demethylation, promoting the proliferation and glycolytic metabolism of EBV+ BL cells, and inhibiting cells apoptosis.