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Submitted on June 26, 2003
Accepted on September 15, 2003
1 Departments of Neurology, and Anatomy & Neurobiology, University of Maryland, School of Medicine, Baltimore MD, 21201
* To whom correspondence should be addressed. E-mail: ckoski{at}umaryland.edu.
Mechanisms underlying the divergent effects of ovarian hormones on neuron death induced by tumor necrosis factor-
(TNF) were investigated in differentiated PC12 cells (dPC12). dPC12 were exposed to 17
-estradiol (E, 1.0 nM), progesterone (P,100 nM) or a combination of both hormones for 0-72 h before treatment with TNF (0-150ng) to induce cell death. Cells undergoing apoptosis were identified by a TUNEL assay and fluorescence activated cell sorting (FACS) after 18 h. Cell death induced by TNF was decreased 89% after E treatment and increased 2-fold after P treatment compared with cells treated with TNF alone. Treatment with E for 24 h before TNF exposure was required for maximum neuroprotection, whereas P-enhanced death was maximal after a 30 min P treatment. TNF induced a 3-fold increased activity of c-JUN-N terminal kinase (JNK1) in d PC12 within 20 min that could be increased 5 to 8-fold by P together with TNF. A peptide inhibitor of JNK1, abrogated P enhancement of TNF-mediated dPC12 death but had only a minimal effect on cell death by TNF alone. Inhibition of caspase 8 activation reduced death induced by TNF alone but was much less effective for P+TNF. P alone did not activate caspase 8. E increased estrogen receptor (ER) and Bcl-xL expression, and all but abolished TNF receptor 1 (TNFR1) expression. P decreased ER and Bcl-xL expression and doubled TNFR1 expression. These data suggest that P regulates apoptosis or survival through augmentation of JNK signaling and altered TNFR1 expression whereas E mainly affects the expression of BCL-xL, TNFR1 and ER.
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