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(The FASEB Journal. 1999;13:69-79.)
© 1999 FASEB

Research Communications

Hydrogen peroxide produced during {gamma}-glutamyl transpeptidase activity is involved in prevention of apoptosis and maintainance of proliferation in U937 cells

Barbara Del Belloa, Aldo Paolicchib, Mario Comportia, Alfonso Pompellaa,1 and Emilia Maellaroa

a Institute of General Pathology, University of Siena Medical School, I-53100 Siena, Italy
b Department of Experimental Biomedicine, University of Pisa Medical School, I-56100 Pisa, Italy

It has been reported in several cell lines that exposure to low levels of reactive oxygen species can exert a stimulatory effect on their proliferation. We have previously shown that mild oxidative conditions can also counteract apoptotic stimuli. A constitutive cellular production of low levels of superoxide and hydrogen peroxide originates from various sources; among these, {gamma}-glutamyl transpeptidase (GGT), the plasma membrane-bound activity in charge of metabolizing extracellular reduced glutathione, has recently been included. Since the inhibition of GGT is a sufficient stimulus for the induction of apoptosis in selected cell lines, we investigated whether this effect might result from the suppression of the mentioned GGT-dependent prooxidant reactions, on the theory that the latter may represent a basal antiapoptotic and proliferative signal for the cell. Experiments showed that: 1) GGT activity in U937 monoblastoid cells is associated with the production of low levels of hydrogen peroxide, and two independent GGT inhibitors cause a dose-dependent decrease of such GGT-dependent production of H2O2; 2) GGT inhibition with acivicin results in cell growth arrest, and induces cell death and DNA fragmentation with the ladder appearance of apoptosis; 3) treatment of cells with catalase—and even more with Trolox C—is able to decrease their proliferative rate; 4) GGT inhibition (with suppression of H2O2 production) results in a down-regulation of poly(ADP-ribose) polimerase (PARP) activity, which precedes the proteolytic cleavage of PARP molecule, such as that typically induced by caspases. The reported data suggest that the low H2O2 levels originating as a by-product during GGT activity are able to act as sort of a `life signal' in U937 cells, insofar as they can maintain cell proliferation and protect against apoptosis, possibly through an up-regulation of PARP activity.—Del Bello, B., Paolicchi, A., Comporti, M., Pompella, A., Maellaro, E. Hydrogen peroxide produced during {gamma}-glutamyl transpeptidase activity is involved in prevention of apoptosis and maintainance of proliferation in U937 cells. FASEB J. 13, 69–79 (1999)


Key Words: oxidative reactions • acivicin • PARP cleavage • poly(ADP-ribose) polymerase




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