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J. Biol. Chem., Vol. 275, Issue 50, 39435-39443, December 15, 2000
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From the Cell Stress and Aging Section, Laboratory of Biological
Chemistry, National Institute on Aging, National Institutes of
Health, Baltimore, Maryland 21224-6825
Cisplatin activates multiple signal transduction
pathways involved in coordinating cellular responses to stress. Here we
demonstrate a requirement for extracellular signal-regulated protein
kinase (ERK), a member of the mitogen-activated protein kinase family in mediating cisplatin-induced apoptosis of human cervical carcinoma HeLa cells. Cisplatin treatment resulted in dose- and time- dependent activation of ERK. That elevated ERK activity contributed to cell death
by cisplatin was supported by several observations: 1) PD98059 and
U0126, chemical inhibitors of the MEK/ERK signaling pathway, prevented
apoptosis; 2) pretreatment of cells with TPA, an activator of the ERK
pathway, enhanced their sensitivity to cisplatin; 3) suramin, a growth
factor receptor antagonist that greatly suppressed ERK activation,
likewise inhibited cisplatin-induced apoptosis; and, finally, 4) HeLa
cell variants selected for cisplatin resistance showed reduced
activation of ERK following cisplatin treatment. Cisplatin-induced
apoptosis was associated with cytochrome c release and
subsequent caspase-3 activation, both of which could be prevented by
treatment with the MEK inhibitors. However, the caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone protected HeLa cells
against apoptosis without affecting ERK activation. Taken together, our
findings suggest that ERK activation plays an active role in mediating
cisplatin-induced apoptosis of HeLa cells and functions upstream of
caspase activation to initiate the apoptotic signal.
Requirement for ERK Activation in Cisplatin-induced
Apoptosis*
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom all correspondence should be addressed: Laboratory of
Biological Chemistry, Box 12, NIA, National Institutes of Health, 5600 Nathan Shock Dr., Baltimore, MD 21224. Tel.: 410-558-8446; Fax:
410-558-8386.
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