The pathway of leukemic cell death caused by glucocorticoid receptor fragment 465*

被引:15
作者
El-Naghy, M
Johnson, BH
Chen, H
Ansari, NH
Zhang, WP
Molle, P
Ji, YS
Thompson, EB [1 ]
机构
[1] Univ Texas, Med Branch, Dept Human Biol Chem & Genet, Galveston, TX 77555 USA
[2] SUNY Stony Brook, Nassau Cty Med Ctr, Dept Internal Med, E Meadow, NY 11554 USA
[3] Univ Penn, Wharton Sch Business, Philadelphia, PA 19103 USA
关键词
apoptosis; caspase-3; glucocorticoid receptor fragment; mitochondria; T-cell leukemia;
D O I
10.1006/excr.2001.5350
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
The truncated glucocorticoid receptor mutant gene 465* codes for a protein that is interrupted by a frame-shift mutation in the second zinc finger of the natural DNA binding domain. Thus, 465* represents the natural amino acid sequence 1-465 followed by 21 novel amino acids starting at position 466. The entire ligand binding domain is missing. Prior studies have shown that transient transfection of the glucocorticoid-resistant leukemic T-cell clone ICR-27 with a plasmid expressing 465* rapidly reduces the number of viable cells. This response does not require activation by a steroid, and a hybrid protein consisting of green fluorescent protein fused to 465* is found primarily in the cytoplasm. In the present study, we present evidence that the decrease in cell number is due to a form of cell death that bears many of the classic characteristics of apoptosis. Expression of the 465* protein can be detected a few hours after electroporation and is followed by activation of caspase-3 as well as reduction of the mitochondrial inner transmembrane potential. The caspase-3 inhibitor ZVAD-fink blocks 465*-dependent cell death when added acutely after electroporation, but fails to do so later. We conclude that the novel 465* gene causes cell death by apoptosis. (C) 2001 Academic Press.
引用
收藏
页码:166 / 175
页数:10
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