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Vol. 25, No. 2-3, 2010
Issue release date: 2010
Cell Physiol Biochem 2010;25:241–252
(DOI:10.1159/000276558)

The Double Edge of Reactive Oxygen Species as Damaging and Signaling Molecules in HL60 Cell Culture

Ferrer M.D. · Sureda A. · Mestre A. · Tur J.A. · Pons A.
Laboratori de Ciències de l’Activitat Física, Departament de Biologia Fonamental i Ciències de la Salut. Grup de Nutrició Comunitaria i Estrés Oxidatiu, IUNICS, Universitat de les Illes Balears, Palma de Mallorca
email Corresponding Author

Abstract

Aims: Our aim was to establish the conditions in which reactive oxygen species produce pathological or hormetic effects on HL60 cells. Methods: HL60 cells were treated with either single bouts (1, 10 and 100 µM) or a sustained production (0.1, 1.0 and 10.0 nM/s) of H2O2. Results: Exposure to 10 and 100 µM H2O2 activated catalase, glutathione peroxidase and glutathione reductase through post-transcriptional mechanisms and induced oxidative modification of proteins. When cells where exposed to sustained H2O2 production, a clear dose-response effect was detected in the activity of the antioxidant enzymes catalase, glutathione peroxidase and Mn-SOD, with higher concentrations of H2O2 inducing greater enzyme activities. Catalase, HO-1, UCP-3, iNOS and PGC-1α expressions were activated after sustained exposure to 1 and 10 nM H2O2/s. Although the antioxidant defenses were activated, oxidative damage appeared in DNA and proteins in cells treated with 1 and 10 nM/s. Conclusions: HL60 cells respond to exposure to sustained levels of H2O2 in a dose-response manner to H2O2 concentration by activating the expression and activity of the antioxidant machinery, although the activation of the antioxidant defenses is not enough to avoid the appearance of oxidative damage. Of the two designs proposed, continuous exposure seems to be more appropriate to study the antioxidant response of HL60 cells to H2O2.


 Outline


 goto top of outline Key Words

  • Adaptive response
  • Gene expression
  • HL60
  • Hormesis
  • Oxidative damage

 goto top of outline Abstract

Aims: Our aim was to establish the conditions in which reactive oxygen species produce pathological or hormetic effects on HL60 cells. Methods: HL60 cells were treated with either single bouts (1, 10 and 100 µM) or a sustained production (0.1, 1.0 and 10.0 nM/s) of H2O2. Results: Exposure to 10 and 100 µM H2O2 activated catalase, glutathione peroxidase and glutathione reductase through post-transcriptional mechanisms and induced oxidative modification of proteins. When cells where exposed to sustained H2O2 production, a clear dose-response effect was detected in the activity of the antioxidant enzymes catalase, glutathione peroxidase and Mn-SOD, with higher concentrations of H2O2 inducing greater enzyme activities. Catalase, HO-1, UCP-3, iNOS and PGC-1α expressions were activated after sustained exposure to 1 and 10 nM H2O2/s. Although the antioxidant defenses were activated, oxidative damage appeared in DNA and proteins in cells treated with 1 and 10 nM/s. Conclusions: HL60 cells respond to exposure to sustained levels of H2O2 in a dose-response manner to H2O2 concentration by activating the expression and activity of the antioxidant machinery, although the activation of the antioxidant defenses is not enough to avoid the appearance of oxidative damage. Of the two designs proposed, continuous exposure seems to be more appropriate to study the antioxidant response of HL60 cells to H2O2.

Copyright © 2010 S. Karger AG, Basel


 goto top of outline Author Contacts

Dr. Antoni Pons Biescas
Laboratori de Ciències de l’Activitat Física
Universitat de les Illes Balears
Ctra. Valldemossa Km 7.5, E-07122 Palma de Mallorca, Illes Balears (Spain)
Tel. +34-971173171, Fax: +34-971173184, E-Mail antonipons@uib.es


 goto top of outline Article Information

Accepted: October 22, 2009
Published online: January 12, 2010
Number of Print Pages : 12


 goto top of outline Publication Details

Cellular Physiology and Biochemistry (International Journal of Experimental Cellular Physiology, Biochemistry and Pharmacology)

Vol. 25, No. 2-3, Year 2010 (Cover Date: 2010)

Journal Editor: F. Lang, Tübingen
ISSN: 1015–8987 (Print), eISSN: 1421–9778 (Online)

For additional information: http://www.karger.com/journals/cpb


Copyright / Drug Dosage / Disclaimer

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

Abstract

Aims: Our aim was to establish the conditions in which reactive oxygen species produce pathological or hormetic effects on HL60 cells. Methods: HL60 cells were treated with either single bouts (1, 10 and 100 µM) or a sustained production (0.1, 1.0 and 10.0 nM/s) of H2O2. Results: Exposure to 10 and 100 µM H2O2 activated catalase, glutathione peroxidase and glutathione reductase through post-transcriptional mechanisms and induced oxidative modification of proteins. When cells where exposed to sustained H2O2 production, a clear dose-response effect was detected in the activity of the antioxidant enzymes catalase, glutathione peroxidase and Mn-SOD, with higher concentrations of H2O2 inducing greater enzyme activities. Catalase, HO-1, UCP-3, iNOS and PGC-1α expressions were activated after sustained exposure to 1 and 10 nM H2O2/s. Although the antioxidant defenses were activated, oxidative damage appeared in DNA and proteins in cells treated with 1 and 10 nM/s. Conclusions: HL60 cells respond to exposure to sustained levels of H2O2 in a dose-response manner to H2O2 concentration by activating the expression and activity of the antioxidant machinery, although the activation of the antioxidant defenses is not enough to avoid the appearance of oxidative damage. Of the two designs proposed, continuous exposure seems to be more appropriate to study the antioxidant response of HL60 cells to H2O2.



 goto top of outline Author Contacts

Dr. Antoni Pons Biescas
Laboratori de Ciències de l’Activitat Física
Universitat de les Illes Balears
Ctra. Valldemossa Km 7.5, E-07122 Palma de Mallorca, Illes Balears (Spain)
Tel. +34-971173171, Fax: +34-971173184, E-Mail antonipons@uib.es


 goto top of outline Article Information

Accepted: October 22, 2009
Published online: January 12, 2010
Number of Print Pages : 12


 goto top of outline Publication Details

Cellular Physiology and Biochemistry (International Journal of Experimental Cellular Physiology, Biochemistry and Pharmacology)

Vol. 25, No. 2-3, Year 2010 (Cover Date: 2010)

Journal Editor: F. Lang, Tübingen
ISSN: 1015–8987 (Print), eISSN: 1421–9778 (Online)

For additional information: http://www.karger.com/journals/cpb


Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.