Augmentation of ethanol-induced cell damage and activation of nuclear factor-kappa B 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)-kappa B 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-kappa B activity was measured by electrophoretic mobility shift assay with a kappa B-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-kappa B activation.


CONCLUSIONS
Ethanol-induced increase in annexin IV expression might amplify ethanol-induced cytotoxicity via NF-kappa B activation.
Sohma
Hitoshi
H
Department of Biochemistry, Sapporo Medical University School of Medicine, Sapporo, Japan. sohma@sapmed.ac.jp
Ohkawa
Hiroko
H
Sakai
Rie
R
Hashimoto
Eri
E
Ukai
Wataru
W
Saito
Toshikazu
T
D000818:Animals; D017314:Annexin A4 / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects*; D024202:Electrophoretic Mobility Shift Assay; D000431:Ethanol / Q000633:toxicity*; D005910:Glioma; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism*; D009447:Neuroblastoma; D051381:Rats; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000187:drug effects*
D017314:Annexin A4; D016328:NF-kappa B; D000431:Ethanol