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Endocrinology Vol. 139, No. 9 3813-3821
Copyright © 1998 by The Endocrine Society

ARTICLES

Glucocorticoid-Induced Apoptosis and Regulation of NF-{kappa}B Activity in Human Leukemic T Cells1

Jyoti Ramdas and Jeffrey M. Harmon

Department of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799

Address all correspondence and requests for reprints to: Jeffrey M. Harmon, Ph.D., Department of Pharmacology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, Maryland 20814-4799. E-mail: jharmon{at}mxb.usuhs.mil

Glucocorticoid-induced apoptosis was investigated in glucocorticoid-sensitive 6TG1.1 and resistant ICR27TK.3 human leukemic T cells. Following glucocorticoid treatment of 6TG1.1 cells, chromatin fragmentation was observed after a delay of 24 h. Fragmentation was not observed in ICR27TK.3 cells containing mutant glucocorticoid receptors (L753F) that are activation-deficient but retain the ability to repress AP-1 activity. Nor was fragmentation observed after treatment with RU38486, indicating that repression of AP-1 activity is not involved. As described in other systems, fragmentation required ongoing protein synthesis. However, inhibition of protein synthesis with cycloheximide anytime during the first 18 h of steroid treatment was as effective in blocking chromatin fragmentation as inhibition for the entire period, suggesting that synthesis of a component with a rapid turnover rate is required. Dexamethasone treatment completely blocked 12-O-tetradecanoylphorbol 13-acetate induction of nuclear factor-{kappa}B (NF-{kappa}B) activity and elicited an increase in the amount of immunoreactive I{kappa}B{alpha} in sensitive 6TG1.1 cells but not in resistant ICR27TK.3 cells. In addition, mild detergent treatment of cell extracts indicated that a substantial amount of cytoplasmic NF-{kappa}B is complexed with I{kappa}B{alpha} or some other inhibitory factor. These results suggest that induction of a labile inhibitory factor such as I{kappa}B{alpha} may contribute to glucocorticoid-induced apoptosis.




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