Ascorbic acid enhances the effects of 6-hydroxydopamine and H2O2 on iron-dependent DNA strand breaks and related processes in the neuroblastoma cell line SK-N-SH.
Neuroblastoma cells accumulate ascorbic acid and iron. It was hypothesized that these features could be exploited for sensitizing neuroblastoma cells for therapy in combination with reactive oxygen intermediates. In the present study the effects of 6-hydroxydopamine (6-OHDA) and H2O2 on metabolic parameters critical for cell survival were investigated in cells with low and high ferritin content in the presence and absence of ascorbate. Human neuroblastoma SK-N-SH cells were pretreated with 100 microM FeSO4 and 10 microM desferrioxamine, respectively, for 24 h yielding cells with different ferritin contents. The effects of 6-OHDA and H2O2 (25 microM-250 microM) in the absence and presence of 1 mM ascorbic acid on DNA strand break formation, activation of poly(ADP-ribose) polymerase, and finally decrease in NAD+ and ATP concentration were investigated. All these parameters were influenced by 6-OHDA and H2O2 in a concentration-dependent manner in a similar way. The effects were most pronounced in ferritin-rich cells and in the presence of ascorbic acid. Using isolated CCC PM2 DNA, 6-OHDA and ascorbic acid caused strand breaks that were prevented in the presence of mannitol or desferrithiocine. H2O2-mediated strand breaks were observed only in the presence of ascorbic acid. Based on these data and data published by others a model explaining the deleterious effects of ascorbic acid on neuroblastoma cells is presented. It is suggested that continuous application of a high dosage of ascorbic acid might be a useful approach in neuroblastoma therapy.
Department of Immunology, Research Institute of Scripps Clinic, La Jolla, California 92037.
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Bruchelt
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Schraufstätter
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Niethammer
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Cochrane
D000255:Adenosine Triphosphate / Q000032:analysis; D001205:Ascorbic Acid / Q000493:pharmacokinetics / Q000494:pharmacology*; D004247:DNA / Q000187:drug effects*; D004249:DNA Damage; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007501:Iron / Q000032:analysis / Q000494:pharmacology*; D009243:NAD / Q000032:analysis; D009447:Neuroblastoma / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D016627:Oxidopamine / Q000494:pharmacology*; D011065:Poly(ADP-ribose) Polymerases / Q000032:analysis; D014407:Tumor Cells, Cultured
D009243:NAD; D016627:Oxidopamine; D000255:Adenosine Triphosphate; D004247:DNA; D006861:Hydrogen Peroxide; D007501:Iron; D011065:Poly(ADP-ribose) Polymerases; D001205:Ascorbic Acid