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A 3D spheroid-specific role of Wnt/β-catenin and Eph-Ephrin in nasopharyngeal carcinoma cells.

One of the greatest unmet needs hindering the successful treatment of nasopharyngeal carcinomas (NPC) is the lack of representative, physiological, and cost-effective models. Although Epstein-Barr virus (EBV) infection is consistently present in NPC, most studies have focused on EBV-negative NPC. For the first time, 3D spheroids model of EBV-positive and -negative NPC cells were established and analyzed as compared to classical 2D culture in various aspects of tumor phenotype and drugs response. Compared to 2D monolayer, 3D spheroids showed a significant increase in their migration capacity, stemness characteristics, hypoxia and drug resistance. Coculture with endothelial cells that mimic the essential interactions in the tumor microenvironment effectively enhanced spheroid dissemination. Furthermore, RNA-sequencing showed significant changes at the transcriptional level compared to 2D monolayer. Particularly, we identified known (VEGF, AKT, mTOR) and novel (Wnt/β-catenin, Eph-Ephrin) cell signaling involved in NPC spheroids. Their targeting using FDA-approved drugs are effective in monoculture and coculture. These findings provide the first evidence on the establishment of an EVB-positive and -negative NPC 3D spheroids in resembling advanced/metastatic features, and the potential in identifying new drug targets.