Faculty Collaboration Database

Doxorubicin-induced apoptosis is associated with increased transcription of endothelial nitric-oxide synthase. Effect of antiapoptotic antioxidants and calcium. J Biol Chem 2001 Dec 14;276(50):47266-76

Date

10/02/2001

Pubmed ID

11579094

DOI

10.1074/jbc.M106829200

Scopus ID

2-s2.0-0035861687 (requires institutional sign-in at Scopus site)   199 Citations

Abstract

The clinical efficacy of the antitumor antibiotic drug doxorubicin (DOX) is severely limited by its dose-limiting cardiotoxicity in cancer patients. DOX-induced generation of reactive oxygen species was proposed to be a major mechanism of its cardiotoxicity. Previously, we showed that DOX undergoes a reductive activation at the reductase domain of endothelial nitric-oxide synthase (eNOS) forming the semiquinone and superoxide (Vásquez-Vivar, J., Martasek, P., Hogg, N., Masters, B. S. S., Pritchard, K. A., Jr., and Kalyanaraman, B. (1997) Biochemistry 36, 11293-11297). In this report, we provide evidence for DOX-induced increase in eNOS transcription and protein expression in bovine aortic endothelial cells (BAEC). We propose that DOX-induced hydrogen peroxide formation is responsible for the increased transcription of eNOS. BAEC treated with antisense eNOS oligonucleotide inhibits DOX-induced endothelial apoptosis. Treatment with antioxidants restored the levels of antiapoptotic proteins (Hsp70 and Bcl-2) in DOX-treated BAEC. DOX-induced intracellular oxidative stress, as measured by oxidation of dichlorodihydrofluorescein diacetate to dichlorofluorescein and hydroethidium to ethidium, was inhibited by antisense eNOS oligonucleotide and antioxidant treatment. Furthermore, antiapoptotic antioxidants (e.g. FeTBAP, ebselen, and alpha-phenyl-tert-butyl nitrone) inhibited DOX-induced eNOS transcription. We conclude that DOX-induced apoptosis is linked to the redox activation of DOX by eNOS.

Author List

Kalivendi SV, Kotamraju S, Zhao H, Joseph J, Kalyanaraman B

Author

Balaraman Kalyanaraman PhD Professor in the Biophysics department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Antineoplastic Agents
Antioxidants
Apoptosis
Calcium
Cattle
Cell Line
Cells, Cultured
Dose-Response Relationship, Drug
Doxorubicin
Endothelium
Endothelium, Vascular
Enzyme Activation
Fluoresceins
HSP70 Heat-Shock Proteins
Hydrogen Peroxide
Microscopy, Fluorescence
Mitochondria
Models, Biological
Models, Chemical
NADPH-Ferrihemoprotein Reductase
Nitrates
Nitric Oxide Synthase
Nitric Oxide Synthase Type II
Nitrites
Oligonucleotides, Antisense
Oxidative Stress
Oxygen
Protein Structure, Tertiary
Proto-Oncogene Proteins c-bcl-2
RNA, Messenger
Reverse Transcriptase Polymerase Chain Reaction
Time Factors
Transcription, Genetic