Carcinogenesis, Vol 18, 295-301, Copyright © 1997 by Oxford University Press
T Muller, HJ Haussmann and G Schepers
Previous studies have shown that exposure of Swiss 3T3 cells to mainstream
cigarette smoke (CS) trapped in phosphate-buffered saline (smoke-bubbled
PBS) resulted in the expression of stress response genes, i.e. haem
oxygenase and c-fos, partial inhibition of protein phosphatases 1 and 2A,
as well as partial depletion of the cellular glutathione (GSH) pool. Using
c-fos gene expression in Swiss 3T3 cells as an indicator for a cellular
response against oxidative stress, the following observations are
consistent with peroxynitrite as an active principal formed by CS in
aqueous solutions: (i) sustained c-fos expression was obtained for
smoke-bubbled PBS, peroxynitrite itself and a compound known to
stoichiometrically release superoxide and nitric oxide (NO)
(3-morpholino-sydnonimine, SIN-1); (ii) c-fos expression in cells exposed
to aqueous smoke fractions was inhibited by either the
superoxide-scavenging enzyme superoxide dismutase (SOD), in combination
with catalase, or the NO-scavenger oxyhaemoglobin (HbO2); and (iii)
activation of guanylate cyclase in rat lung cells was observed only when
bubbling was performed with filtered smoke and with whole smoke in the
presence of SOD/catalase. These results are consistent with a rapid
NO-consuming reaction coupled with superoxide-generating properties of the
particulate phase of CS. Moreover, (iv) the half-life of the c-fos-
inducing activity in smoke-bubbled PBS was found to be <1 h which can be
explained by a sustained peroxynitrite formation. Finally, depletion of
intracellular thiol levels by smoke-bubbled PBS appears to favour the
activation of a redox-sensitive component of the c-fos-inducing pathway.
ARTICLES
Evidence for peroxynitrite as an oxidative stress-inducing compound of aqueous cigarette smoke fractions
INBIFO Institut fur biologische Forschung GmBH, Koln, Germany.
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