Hsp90 cleavage by an oxidative stress leads to its client proteins degradation and cancer cell death.
The heat shock protein 90 (Hsp90) plays a crucial role in the stability of several proteins that are essential for malignant transformation. Hsp90 is therefore an interesting therapeutic target for cancer therapy. In this paper, we investigated whether an oxidative stress generated during ascorbate-driven menadione redox cycling (ascorbate/menadione), affects Hsp90 leading to the degradation of some critical proteins and cell death. Unlike 17-AAG, which inhibits Hsp90 but enhances Hsp70 levels, ascorbate/menadione-treated cells present an additional Hsp90 protein band of about 70kDa as shown by Western blot analysis, suggesting Hsp90 cleavage. This Hsp90 cleavage seems to be a selective phenomenon since it was observed in a large panel of cancer cell lines but not in non-transformed cells. Antibodies raised against either the N-terminus or the C-terminus domains of Hsp90 suggest that the site of cleavage should be located at its N-terminal part. Furthermore, antibodies raised against either the alpha- or the beta-Hsp90 isoform show that Hsp90beta is cleaved while the alpha isoform is down-regulated. We have further shown that different Hsp90 client proteins like Bcr-Abl (a chimerical protein expressed in K562 leukemia cells), RIP and Akt, were degraded when K562 cells were exposed to an oxidative stress. Both Hsp90 cleavage and Bcr-Abl degradation were observed by incubating K562 cells with another H(2)O(2)-generating system (glucose/glucose oxidase) and by incubating KU812 cells (another leukemia cell line) with ascorbate/menadione. Due to the major role of Hsp90 in stabilizing oncogenic and mutated proteins, these results may have potential clinical applications.
Beck
Raphael
R
Unité de Pharmacocinétique, Métabolisme, Nutrition, et Toxicologie, Département des sciences pharmaceutiques, Université Catholique de Louvain, avenue E. Mounier 73, 1200 Brussels, Belgium.
Verrax
Julien
J
Gonze
Thomas
T
Zappone
Marianne
M
Pedrosa
Rozangela Curi
RC
Taper
Henryk
H
Feron
Olivier
O
Calderon
Pedro Buc
PB
D017209:Apoptosis; D001205:Ascorbic Acid / Q000494:pharmacology; D015153:Blotting, Western; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D020014:K562 Cells; D018384:Oxidative Stress; D024483:Vitamin K 3 / Q000494:pharmacology
D018841:HSP90 Heat-Shock Proteins; D024483:Vitamin K 3; D001205:Ascorbic Acid