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Vol. 58, Issue 3, 535-541, September 2000
Laboratory of Membrane Biochemistry and Biophysics, National
Institute on Alcohol Abuse and Alcoholism, Rockville, Maryland (Y.S.,
K.-S.J., I.J.L., M.-A.B., B.J.S.); and Laboratory of Bioorganic
Chemistry, National Institute of Diabetes and Digestive and Kidney
Diseases, Bethesda, Maryland (Y.-C.K.)
The by-product of lipid peroxidation, 4-hydroxynonenal (HNE), was shown
to cause apoptosis in PC12 cells. In this study, we investigated the
molecular mechanism of HNE-induced apoptosis in these cells.
Specifically, we determined the effect of HNE on the activities of
mitogen-activated protein (MAP) kinases involved in early signal
transduction. Within 15 to 30 min after HNE treatment, c-Jun N-terminal
protein kinase (JNK) was maximally activated, before it returned to
control level at 1 h post-treatment. In contrast, activities of
extracellular signal-regulated kinase and p38 MAP kinase
remained unchanged from their baseline levels. Stress-activated protein
kinase kinase (SEK1), an upstream kinase of JNK, was also activated
within 5 min after HNE treatment and remained activated for up to 60 min. Marked activation of the JNK pathway through SEK1 and apoptosis
signal-regulating kinase 1 (ASK1), an upstream kinase of SEK1, was
demonstrated by the transient transfection of cDNA for wild-type SEK1
or ASK1 together with JNK into COS-7 cells. Furthermore, significant
reductions in JNK activation and HNE-induced cell death were observed
when either of the dominant negative mutant of SEK1 or ASK1 was
cotransfected with JNK. Pretreatment of PC12 cells with a
survival-promoting agent, 8-(4-chlorophenylthio)-cAMP, prevented both
the HNE-induced JNK activation and apoptosis. Nonaldehyde, a nontoxic
aldehyde, neither caused apoptosis nor JNK activation. Pretreatment of
PC12 cells with SB203580, a specific inhibitor of p38 MAP kinase, had no effect on HNE-induced apoptosis. All these data suggest that the
selective JNK activation by HNE is critical for the apoptosis of PC12
cells and that the HNE-mediated apoptosis is likely to be mediated
through the activation of the ASK1-SEK1-JNK pathway without
activation of extracellular signal-regulated kinase or p38 MAP kinase.
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