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(Received for publication, January 11, 1996, and in revised form, May 21, 1996)
mer, John C.
Reed
and
From the University of Erlangen Nürnberg, Faculty of
Medicine, Department of Medicine IV-Experimental Division,
Loschgestra Endogenously generated or exogenously supplied
nitric oxide (NO)-induced apoptotic cell death in the mouse macrophage
cell line RAW 264.7. Apoptotic signaling caused an early accumulation
of the tumor suppressor p53 prior to DNA fragmentation. Contrary to the
notion of specific activating signals, inhibitory transduction
mechanisms largely remain unknown. Therefore, RAW 264.7 macrophages
were stably transfected with human Bcl-2, an anti-apoptotic protein.
Bcl-2 transfectants showed substantial protection from cell death
induced following the exposure to NO donors such as
S-nitrosoglutathione (GSNO) and spermine-NO. In
contrast, in RAW 264.7 parent or in neomycin control-transformed cells,
these NO donors induced internucleosomal DNA cleavage in a
dose-dependent manner. Similarly, expression of the
inducible NO synthase in response to lipopolysaccharide and
interferon-e 8, 91054 Erlangen, Federal Republic of Germany and the
The Burnham Institute, Cancer Research Center, La
Jolla, California 92037
also caused apoptosis in RAW macrophages and
neo controls within 24 h. In contrast, Bcl-2
transfectants appeared highly resistant, although inducible NO synthase
levels increased along with concomitant nitrite production similar to
control cells. The expression of p53 and Bax was also explored in
controls and Bcl-2 transfectants after GSNO addition. GSNO induced p53
expression in Bcl-2 transfectants at levels comparable with
nontransfected RAW macrophages. Moreover, GSNO induced increases in
the steady-state levels of Bax protein in parental and
Bcl-2-transfected cells. We conclude therefore, that Bcl-2 acts
downstream of p53, presumably nullifying the NO-mediated increase in
Bax protein in RAW 264.7 cells.
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B. Brune, C. Gotz, U. K. Messmer, K. Sandau, M.-R. Hirvonen, and E. G. Lapetina Superoxide Formation and Macrophage Resistance to Nitric Oxide-mediated Apoptosis J. Biol. Chem., March 14, 1997; 272(11): 7253 - 7258. [Abstract] [Full Text] [PDF]
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L. Piacenza, G. Peluffo, and R. Radi L-Arginine-dependent suppression of apoptosis in Trypanosoma cruzi: Contribution of the nitric oxide and polyamine pathways PNAS, June 19, 2001; 98(13): 7301 - 7306. [Abstract] [Full Text] [PDF]
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