Activation of nuclear factor-kappaB during doxorubicin-induced apoptosis in endothelial cells and myocytes is pro-apoptotic: the role of hydrogen peroxide.

Doxorubicin (DOX) is a widely used anti-tumour drug. Cardiotoxicity is a major toxic side effect of DOX therapy. Although recent studies implicated an apoptotic pathway in DOX-induced cardiotoxicity, the mechanism of DOX-induced apoptosis remains unclear. In the present study, we investigated the role of reactive oxygen species and the nuclear transcription factor nuclear factor kappaB (NF-kappaB) during apoptosis induced by DOX in bovine aortic endothelial cells (BAECs) and adult rat cardiomyocytes. DOX-induced NF-kappaB activation is both dose- and time-dependent, as demonstrated using electrophoretic mobility-shift assay and luciferase and p65 (Rel A) nuclear-translocation assays. Addition of a cell-permeant iron metalloporphyrin significantly suppressed NF-kappaB activation and apoptosis induced by DOX. Overexpression of glutathione peroxidase, which detoxifies cellular H(2)O(2), significantly decreased DOX-induced NF-kappaB activation and apoptosis. Inhibition of DOX-induced NF-kappaB activation by a cell-permeant peptide SN50 that blocks translocation of the NF-kappaB complex into the nucleus greatly diminished DOX-induced apoptosis. Apoptosis was inhibited when IkappaB mutant vector, another NF-kappaB inhibitor, was added to DOX-treated BAECs. These results suggest that NF-kappaB activation in DOX-treated endothelial cells and myocytes is pro-apoptotic, in contrast with DOX-treated cancer cells, where NF-kappaB activation is anti-apoptotic. Removal of intracellular H(2)O(2) protects endothelial cells and myocytes from DOX-induced apoptosis, possibly by inhibiting NF-kappaB activation. These findings suggest a novel mechanism for enhancing the therapeutic efficacy of DOX.

[1]  A. Porter,et al.  NF-κB Activation Mediates Doxorubicin-induced Cell Death in N-type Neuroblastoma Cells* , 2001, The Journal of Biological Chemistry.

[2]  J. Joseph,et al.  Doxorubicin-induced Apoptosis Is Associated with Increased Transcription of Endothelial Nitric-oxide Synthase , 2001, The Journal of Biological Chemistry.

[3]  J. Camp,et al.  Reactivity of mono-meso-substituted iron(II) octaethylporphyrin complexes with hydrogen peroxide in the absence of dioxygen. Evidence for nucleophilic attack on the heme. , 2001, Journal of the American Chemical Society.

[4]  J. Jaffrezou,et al.  Signaling pathways activated by daunorubicin. , 2001, Blood.

[5]  M. Aoki,et al.  Endothelial Apoptosis Induced by Oxidative Stress Through Activation of NF-&kgr;B: Antiapoptotic Effect of Antioxidant Agents on Endothelial Cells , 2001, Hypertension.

[6]  S. Li,et al.  GPx-1 gene delivery modulates NFkappaB activation following diverse environmental injuries through a specific subunit of the IKK complex. , 2001, Antioxidants & redox signaling.

[7]  L. Marnett,et al.  IκB Kinase, a Molecular Target for Inhibition by 4-Hydroxy-2-nonenal* , 2001, The Journal of Biological Chemistry.

[8]  C. Wanner,et al.  Oxidized LDL suppresses NF-kappaB and overcomes protection from apoptosis in activated endothelial cells. , 2001, Journal of the American Society of Nephrology : JASN.

[9]  H. Kalthoff,et al.  Inhibition of NF-κB sensitizes human pancreatic carcinoma cells to apoptosis induced by etoposide (VP16) or doxorubicin , 2001, Oncogene.

[10]  G. Pieper,et al.  Antioxidant treatment inhibits activation of myocardial nuclear factor kappa B and inhibits nitrosylation of myocardial heme protein in cardiac transplant rejection. , 2001, Antioxidants & redox signaling.

[11]  J. Joseph,et al.  Bicarbonate exacerbates oxidative injury induced by antitumor antibiotic doxorubicin in cardiomyocytes. , 2000, American journal of physiology. Heart and circulatory physiology.

[12]  J. Joseph,et al.  Doxorubicin-induced Apoptosis in Endothelial Cells and Cardiomyocytes Is Ameliorated by Nitrone Spin Traps and Ebselen , 2000, The Journal of Biological Chemistry.

[13]  B. Aggarwal.  Apoptosis and nuclear factor-κb: a tale of association and dissociation , 2000 .

[14]  Y. Ueda,et al.  Fas-mediated apoptosis in adriamycin-induced cardiomyopathy in rats: In vivo study. , 2000, Circulation.

[15]  M. Runge,et al.  Doxorubicin-Induced Cardiomyopathy , 2000 .

[16]  Y Li,et al.  [Mitochondria and apoptosis]. , 2000, Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine].

[17]  J. Crow,et al.  Manganese and iron porphyrins catalyze peroxynitrite decomposition and simultaneously increase nitration and oxidant yield: implications for their use as peroxynitrite scavengers in vivo. , 1999, Archives of biochemistry and biophysics.

[18]  C. Der,et al.  Integration of Rac-dependent Regulation of Cyclin D1 Transcription through a Nuclear Factor-κB-dependent Pathway* , 1999, The Journal of Biological Chemistry.

[19]  Hsiu-Hsiang Lee,et al.  NF‐κB‐dependent Fas ligand expression , 1999 .

[20]  M. Baudry,et al.  EUK-134, a synthetic superoxide dismutase and catalase mimetic, prevents oxidative stress and attenuates kainate-induced neuropathology. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[21]  T. Chase,et al.  Nuclear Factor κB Nuclear Translocation Upregulates c-Myc and p53 Expression during NMDA Receptor-Mediated Apoptosis in Rat Striatum , 1999, The Journal of Neuroscience.

[22]  N. Olashaw,et al.  Tumor Necrosis Factor-α-induced Proliferation of Human Mo7e Leukemic Cells Occurs via Activation of Nuclear Factor κB Transcription Factor* , 1999, The Journal of Biological Chemistry.

[23]  H. Jo,et al.  Effects of pyrrolidine dithiocarbamate on endothelial cells: protection against oxidative stress. , 1999, Free radical biology & medicine.

[24]  L. Owen-Schaub,et al.  Activation-Dependent Transcriptional Regulation of the Human fas Promoter Requires NF-κB p50-p65 Recruitment , 1999, Molecular and Cellular Biology.

[25]  D. Sawyer,et al.  Daunorubicin-induced apoptosis in rat cardiac myocytes is inhibited by dexrazoxane. , 1999, Circulation research.

[26]  S. Manna,et al.  Lipopolysaccharide inhibits TNF-induced apoptosis: role of nuclear factor-kappaB activation and reactive oxygen intermediates. , 1999, Journal of immunology.

[27]  M. Karin,et al.  An Essential Role for Nuclear Factor κB in Promoting Double Positive Thymocyte Apoptosis , 1999, The Journal of experimental medicine.

[28]  Lei Wang,et al.  Regulation of cardiomyocyte apoptotic signaling by insulin-like growth factor I. , 1998, Circulation research.

[29]  M. Currie,et al.  Characterization of the Cytoprotective Action of Peroxynitrite Decomposition Catalysts* , 1998, The Journal of Biological Chemistry.

[30]  M. Currie,et al.  Peroxynitrite decomposition catalysts: therapeutics for peroxynitrite-mediated pathology. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[31]  J. Marshall,et al.  Pyrrolidine dithiocarbamate attenuates endotoxin-induced acute lung injury. , 1997, American journal of respiratory cell and molecular biology.

[32]  N. Thornberry,et al.  Caspases: killer proteases. , 1997, Trends in biochemical sciences.

[33]  L. O’Neill,et al.  Daunorubicin Activates NFκB and Induces κB-dependent Gene Expression in HL-60 Promyelocytic and Jurkat T Lymphoma Cells* , 1997, The Journal of Biological Chemistry.

[34]  P. Golstein.  Controlling Cell Death , 1997, Science.

[35]  S. Liochev,et al.  A metalloporphyrin superoxide dismutase mimetic protects against paraquat-induced endothelial cell injury, in vitro. , 1995, The Journal of pharmacology and experimental therapeutics.

[36]  J. Ye,et al.  Characterization of a silencer regulatory element in the human interferon-gamma promoter. , 1994, The Journal of biological chemistry.

[37]  S. Ben‐Sasson,et al.  Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation , 1992, The Journal of cell biology.

[38]  J. Doroshow,et al.  Redox cycling of anthracyclines by cardiac mitochondria. I. Anthracycline radical formation by NADH dehydrogenase. , 1986, The Journal of biological chemistry.

[39]  A. Buzdar,et al.  Early and delayed clinical cardiotoxicity of doxorubicin , 1985, Cancer.

[40]  R. Mason,et al.  Spin-trapping and direct electron spin resonance investigations of the redox metabolism of quinone anticancer drugs. , 1980, Biochimica et biophysica acta.

[41]  I. Ifrim,et al.  Adriamycin: the role of lipid peroxidation in cardiac toxicity and tumor response. , 1977, Science.

[42]  R. Beyaert.  Nuclear Factor кB , 2003, Springer Netherlands.

[43]  J. G. Cory,et al.  Enhanced roscovitine-induced apoptosis is mediated by a caspase-3-like activity in deoxyadenosine-resistant mouse leukemia L1210 cells. , 2000, Anticancer research.

[44]  Xianglin Shi,et al.  New insights into the role of nuclear factor-kappaB, a ubiquitous transcription factor in the initiation of diseases. , 1999, Clinical chemistry.

[45]  P. Cerutti,et al.  The roles of hydrogen peroxide and superoxide as messengers in the activation of transcription factor NF-kappa B. , 1995, Chemistry & biology.