Augmentation of ethanol-induced cell damage and activation of nuclear factor-κB by annexin IV in cultured cells

Background: We previously reported that the amount of annexin IV expression was increased in culture cells by exposure to ethanol. Here, to investigate the physiologic role of annexin IV, we analyzed ethanol-induced cytotoxicity and nuclear factor (NF)-kappaB activity by using annexin IV-overexpressed cells. Methods: Annexin IV overexpression was performed by transfection of expression vector, in which annexin IV complementary DNA was ligated, to culture cells (rat glioma C6 cells and human neuroblastoma SH-SY5Y cells). Ethanol-induced cytotoxicity was assayed by measuring the mitochondrial enzyme (dehydrogenase) activity or trypan blue exclusion. NF-kappaB activity was measured by electrophoretic mobility shift assay with a kappaB-oligonucleotide probe. Results: Ethanol-induced cytotoxicity was increased by overexpression of annexin IV in both C6 cells and SH-SY5Y cells. Annexin IV overexpression augmented ethanol-induced NF-kappaB activation. Conclusions: Ethanol-induced increase in annexin IV expression might amplify ethanol-induced cytotoxicity via NF-kappaB activation.