Arachidonate release and c-fos expression in various models of hypoxia and hypoxia-hypoglycemia in retinoic acid differentiated neuroblastoma cells.
Hypoxia-hypoglycemia has played an important role in inducing both phospholipase A2 activation and the expression of the early gene c-fos, in the neuroblastoma cell line SK-N-BE, after it has been differentiated by retinoic acid. Under hypoxic-hypoglycemic conditions, arachidonic acid release has found to be significant after 30 min, whereas c-fos expression has required at least 4 h. This model has been obtained by adding glycolytic inhibitor 2-deoxyglucose to the culture and by placing cells in an atmosphere containing 100% N2 for different time periods. This condition has been compared with two different models: NaCN and nitrogen have been used as hypoxic stimuli, without inhibiting the glycolytic pathway, but the same cell cultures have been used. Cell viability and the fall of cellular ATP levels have been evaluated in all the models, in order to monitor and compare the hypoxic cellular damage. Phospholipase A2 activation has been found to be significant in all conditions, even if to a different extent; but only hypoxia combined with the inhibition of the glycolytic pathway, has induced a significant expression of c-fos. It is very difficult to study hypoxic stimuli in 'in vitro' systems. Our study has compared three different models and the one combining gaseous hypoxia and hypoglycemic conditions seems to be very effective in stimulating early events involved in hypoxic phenomena such as phospholipase activation and the expression of the early gene c-fos.
Petroni
A
A
Institute of Pharmacological Sciences, University of Milan, via Balzaretti 9, 20133, Milan, Italy. anna.petroni@unimi.it
Papini
N
N
Blasevich
M
M
Rise
P
P
Galli
C
C
D016718:Arachidonic Acid / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D004789:Enzyme Activation / Q000502:physiology; D006019:Glycolysis / Q000502:physiology; D006801:Humans; D007003:Hypoglycemia / Q000150:complications* / Q000378:metabolism*; D000860:Hypoxia / Q000139:chemically induced / Q000150:complications* / Q000378:metabolism*; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology; D009584:Nitrogen; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism*; D012966:Sodium Cyanide; D013997:Time Factors; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D016760:Proto-Oncogene Proteins c-fos; D016718:Arachidonic Acid; D014212:Tretinoin; D010741:Phospholipases A; D054467:Phospholipases A2; D009584:Nitrogen; D012966:Sodium Cyanide