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J Biol Chem, Vol. 274, Issue 39, 27545-27552, September 24, 1999
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From the Mitogen-activated protein kinase (MAPK) cascades
are activated by diverse extracellular signals and participate in the
regulation of an array of cellular programs. In this study, we
investigated the roles of MAPKs in the induction of phase II
detoxifying enzymes by chemicals. Treatment of human hepatoma (HepG2)
and murine hepatoma (Hepa1c1c7) cells with
tert-butylhydroquinone (tBHQ) or sulforaphane (SUL), two
potent phase II enzyme inducers, stimulated the activity of
extracellular signal-regulated protein kinase 2 (ERK2) but not c-Jun
N-terminal kinase 1. tBHQ and SUL also activated MAPK kinase.
Inhibition of MAPK kinase with its inhibitor, PD98059, abolished ERK2
activation and impaired the induction of quinone reductase, a phase II
detoxifying enzyme, and antioxidant response element (ARE)-linked
reporter gene by tBHQ and SUL. Overexpression of a dominant-negative
mutant of ERK2 also attenuated tBHQ and SUL induction of ARE reporter
gene activity. Interestingly, although expression of Ras and its mutant
forms showed distinct effects on basal ARE reporter gene activity, they
did not affect the activation of reporter gene by the inducers.
Furthermore, a dominant-negative mutant of Ras had little effect on
ERK2 activation by tBHQ and SUL, implicating a Ras-independent
mechanism. Indeed, both tBHQ and SUL were able to stimulate Raf-1
kinase activity in vivo as well as in vitro.
Thus, our results indicate that the induction of
ARE-dependent phase II detoxifying enzymes is mediated by a MAPK pathway, which may involve direct activation of Raf-1 by the inducers.
Department of Pharmaceutics and
Pharmacodynamics, Center for Pharmaceutical Biotechnology, College of
Pharmacy, University of Illinois at Chicago, Chicago, Illinois 60612, the § Department of Microbiology, University of Virginia,
Charlottesville, Virginia 22908, the ¶ Department of Pharmacology,
Lineberger Comprehensive Cancer Center, University of North Carolina,
Chapel Hill, North Carolina 27599, and the
H. Lee Moffitt Cancer
Center and Department of Medical Microbiology and Immunology,
University of South Florida, Tampa, Florida 33612
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