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Pulmonary and Critical Care Division, Department of Medicine, New England Medical Center/Tupper Research Institute; and
* Department of Biochemistry, Tufts University School of Medicine, Boston, Massachusetts 02111, USA
1Correspondence: Pulmonary and Critical Care Division, New England Medical Center, 750 Washington St., NEMC #257, Boston, MA 02111, USA. E-mail vthannickal{at}lifespan.org
Mitogenic growth factors and transforming growth factor ß1 (TGF-ß1)
induce the generation of reactive oxygen species (ROS) in
nonphagocytic cells, but their enzymatic source(s) and regulatory
mechanisms are largely unknown. We previously reported on the ability
of TGF-ß1 to activate a cell surface-associated
NADH:flavin:O2 oxidoreductase (NADH oxidase) that generates
extracellular H2O2. In this study, we compared
the ROS-generating enzymatic systems activated by mitogenic growth
factors and TGF-ß1 with respect to the primary reactive species
produced (O2.- vs.
H2O2), the site of generation (intracellular
vs. extracellular) and regulation by Ras. We find that the mitogenic
growth factors PDGF-BB, FGF-2, and TGF-
(an EGF receptor ligand) are
able to rapidly (within 5 min) induce the generation of intracellular
O2.- without detectable NADH oxidase activity
or extracellular H2O2 release. In contrast,
TGF-ß1 does not stimulate intracellular O2.-
production and the delayed induction of extracellular
H2O2 release is not associated with
O2.- production. Expression of
dominant-negative Ras (N17Ras) protein by herpes simplex virus-mediated
gene transfer blocks mitogen-stimulated intracellular
O2.- generation but has no effect on
TGF-ß1-induced NADH oxidase activation/H2O2
production. These results demonstrate that there are at least two
distinctly different ROS-generating enzymatic systems in lung
fibroblasts regulated by mitogenic growth factors and TGF-ß1 via
Ras-dependent and -independent mechanisms, respectively. In addition,
these findings suggest that endogenous production of ROS by growth
factors/cytokines may have different biological effects depending on
the primary reactive species generated and site of
production.—Thannickal, V. J., Day, R. M., Klinz, S. G., Bastien, M. C., Larios, J. M., Fanburg, B. L.
Ras-dependent and -independent regulation of reactive oxygen species by
mitogenic growth factors and TGF-ß1.
Key Words: superoxide anion • hydrogen peroxide • cell growth • fibroblasts
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