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(The FASEB Journal. 1999;13:2061-2070.)
© 1999 FASEB

HSP27 inhibits cytochrome c-dependent activation of procaspase-9

CARMEN GARRIDO1, JEAN-MARIE BRUEY, ANNIE FROMENTIN, ARLETTE HAMMANN, ANDRÉ PATRICK ARRIGO* and ERIC SOLARY

INSERM U517, Groupe Biologie et Thérapie des Cancers (JE 515), Faculty of Medicine and Pharmacy, 21033 Dijon, France; and
* Stress Laboratory, CNRS UMR-5534, Claude Bernard University, Lyon, France

1Correspondence: INSERM U517, Faculty of Medicine and Pharmacy, 7 Blvd. Jeanne D'Arc, 21033 Dijon, France. E-mail: cgarrido{at}u-bourgogne.fr

We have previously shown that the small heat shock protein HSP27 inhibited apoptotic pathways triggered by a variety of stimuli in mammalian cells. The present study demonstrates that HSP27 overexpression decreases U937 human leukemic cell sensitivity to etoposide-induced cytotoxicity by preventing apoptosis. As observed for Bcl-2, HSP27 overexpression delays poly(ADP-ribose)polymerase cleavage and procaspase-3 activation. In contrast with Bcl-2, HSP27 overexpression does not prevent etoposide-induced cytochrome c release from the mitochondria. In a cell-free system, addition of cytochrome c and dATP to cytosolic extracts from untreated cells induces the proteolytic activation of procaspase-3 in both control and bcl-2-transfected U937 cells but fails to activate procaspase-3 in HSP27-overexpressing cells. Immunodepletion of HSP27 from cytosolic extracts increases cytochrome c/dATP-mediated activation of procaspase-3. Overexpression of HSP27 also prevents procaspase-9 activation. In the cell-free system, immunodepletion of HSP27 increases LEDH-AFC peptide cleavage activity triggered by cytochrome c/dATP treatment. We conclude that HSP27 inhibits etoposide-induced apoptosis by preventing cytochrome c and dATP-triggered activity of caspase-9, downstream of cytochrome c release.—Garrido, C., Bruey, J.-M., Fromentin, A., Hammann, A., Arrigo, A. P., Solary, E. HSP27 inhibits cytochrome c-dependent activation of procaspase-9.


Key Words: apoptosis • cell death • etoposide • drug resistance • leukemia




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