Overexpression of EphA2 Promotes the Establishment of Lung Cancer Cell Line With Stable Latent EB Virus Infection

Objective To establish an Epstein-Barr virus (EBV) infectious lung cancer cells mediated by EphA2 overexpression.

Methods A lentiviral system was employed to establish a stable cell line with ectopic overexpression of EphA2 in lung cancer cell line. These engineered cells were subsequently co-cultured with Akata B cells which carry EBV-eGFP. Following co-culture, successfully infected epithelial cells were selected using G418 and validated by immunofluorescence of EBNA1. Phenotypic assays, including colony formation, wound healing, transwell migration, and invasion assays, were then performed on the established A549-EphA2-EBV cell line to assess the effects of EBV infection.

Results It is difficult to achieve EBV infection in a short period of time through direct contact co-culture of A549 parental cells with Akata-EBV-eGFP. EphA2 overexpression significantly enhanced the infection efficiency of EBV in lung cancer cells, achieving an infection efficiency of 0.53%. Green fluorescent protein expression was observed after more than ten passages of culture, and EBER in situ hybridization confirmed the persistence of EBV infection, demonstrating that the cell line harbors EBV in a stable latent state.

Conclusions We established a lung cancer epithelial cell line with stable latent EBV infection by EphA2 overexpressing, providing essential material for investigating the mechanisms underlying pulmonary lymphoepithelioma carcinoma. The malignant phenotypes, including proliferation and migration, of these latently EBV-infected lung cancer epithelial cells were suppressed, and the underlying molecular mechanisms warrant further investigation.