Potential dengue virus-triggered apoptotic pathway in human neuroblastoma cells: Arachidonic acid, superoxide anion, and NF-kappa B are sequentially involved

Direct in vivo evidence for the susceptibility of human neuronal cells to d engue virus has not been reported. In this study, we demonstrated that type 2 dengue (DEN-2) virus infection induced extensive apoptosis in the human neuroblastoma cell line SK-N-SH. Phospholipase A(2) (PLA(2)) was activated by DEN-2 infection, which led to the generation of arachidonic acid (AA). I nhibition of PLA(2) activity by the PLA(2) inhibitors, AACOCF(3) and ONO-RS -082, diminished DEN-2 virus-induced apoptosis. In contrast, the cyclooxyge nase inhibitors aspirin and indomethacin, thought to increase AA accumulati on by blocking AA catabolism, enhanced apoptosis. Exogenous AA induced apop tosis in a dose-dependent manner. Superoxide anion, which is thought to be generated through the AA-activated NADPH oxidase, was increased after infec tion. Pretreatment with superoxide dismutase (SOD) protected cells against DEN-2 virus-induced apoptosis. Furthermore, generation of superoxide anion was blocked by AACOCF(3). In addition, the transcription factors, NF-kappa B and c-Jun, were found to be activated after DEN-2 virus infection. Howeve r, pretreatment of cells with oligodeoxynucleotides containing NF-kappa B, but not c-Jun, binding sites (transcription factor decoy) strongly prevente d dengue virus-induced apoptosis. The finding that AACOCF(3) and SOD signif icantly block activation of NF-kappa B suggests that this activation is der ived from the AA-superoxide anion pathway. Our results indicate that DEN-2 virus infection of human neuroblastoma cells triggers an apoptotic pathway through PLA(2) activation to superoxide anion generation and subsequently t o NF-kappa B activation. This apoptotic effect can be either directly deriv ed from the action of AA and superoxide anion on mitochondria or indirectly derived from the products of apoptosis-related genes activated by NF-kappa B.