The Epstein-Barr Virus-Encoded MicroRNA MiR-BART9 Promotes Tumor Metastasis by Targeting E-Cadherin in Nasopharyngeal Carcinoma

MicroRNAs (miRNAs) are a family of small RNA molecules that negatively regulate the expression of protein-coding genes and play critical roles in orchestrating diverse cellular processes. This regulatory mechanism is also exploited by viruses to direct their life cycle and evade the host immune system. Epstein-Barr virus (EBV) is an oncogenic virus that is closely associated with multiple human diseases, including nasopharyngeal carcinoma (NPC), which is a highly metastatic type of tumor and is frequently reported in South Asia. Several viral proteins have been found to promote the migration and invasiveness of NPC cells. However, not all tumor tissues express these viral oncoproteins, suggesting that other mechanisms may contribute to the aggressive behavior of NPC tumor cells. A previous sequencing study by our group revealed that the EBV miRNA miR-BART9 was expressed at high levels in all EBV-positive NPC tissues. In the present study, we used gain- and loss-of-function approaches to investigate the effect of miR-BART9 in EBV-negative and EBV-positive NPC cells. We discovered that miR-BART9 promotes the migration and invasiveness of cultured NPC cells. The promigratory activity observed in vitro was manifested as an enhanced metastatic ability in vivo. Computational analysis revealed that miR-BART9 may target E-cadherin, a membrane protein that is pivotal in preserving cell-cell junctions and the epithelial phenotype. Through biochemical assays and functional rescue analysis, we confirmed that miR-BART9 specifically inhibits E-cadherin to induce a mesenchymal-like phenotype and promote the migration of NPC cells. These results indicated that miR-BART9 is a prometastatic viral miRNA and suggested that high levels of miR-BART9 in EBV-positive NPC cells may contribute to the aggressiveness of tumor cells. Author Summary MicroRNAs (miRNAs) are a family of small RNA molecules that negatively regulate the expression of protein-coding genes and orchestrate diverse cellular processes. This regulatory mechanism is also exploited by viruses to manage their life cycle and to evade the host immune system. Epstein-Barr virus (EBV) is closely associated with nasopharyngeal carcinoma (NPC), which is a highly metastatic type of tumor. A previous by our group study discovered that the EBV miRNA miR-BART9 is expressed at high levels in all EBV-positive NPC tissues. In the present study, we used gain- and loss-of-function approaches to investigate the function of miR-BART9 in EBV-negative and EBV-positive NPC cells. We showed that miR-BART9 promotes the migration and invasiveness of cultured NPC cells and enhances the metastatic ability of NPC tumors in vivo. Through computational analysis, we discovered that E-cadherin, a membrane protein that is critical for maintaining cell-cell contact and suppressing tumor metastasis, is a potential target for miR-BART9. Through biochemical assays and a functional rescue analysis, we confirmed that miR-BART9 specifically inhibits E-cadherin to enhance the motility of NPC cells. Our results show that miR-BART9 is a prometastatic viral miRNA and suggest that miR-BART9 may contribute to the aggressiveness of EBV-positive NPC tumors.