This Article Full Text Full Text (PDF) Methods Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Arai, M. Articles by Nagai, R. Search for Related Content PubMed PubMed Citation Articles by Arai, M. Articles by Nagai, R. Related Collections Gene expression Physiological and pathological control of gene expression

(Circulation Research. 2000;86:8.)
© 2000 American Heart Association, Inc.

Molecular Medicine

Mechanism of Doxorubicin-Induced Inhibition of Sarcoplasmic Reticulum Ca²⁺-ATPase Gene Transcription

Masashi Arai, Akemi Yoguchi, Takako Takizawa, Tomoyuki Yokoyama, Tsugiyasu Kanda, Masahiko Kurabayashi, Ryozo Nagai

From the Second Department of Internal Medicine, Gunma University School of Medicine, Gunma, Japan.

Correspondence to Masashi Arai, MD, Second Department of Internal Medicine, Gunma University School of Medicine, Showa-Machi 3-39-22, Maebashi, Gunma 371-8511, Japan. E-mail araim{at}akagi.sb.gunma-u.ac.jp

Abstract—Doxorubicin (DOX)–induced cardiomyopathy has been found to be associated with impaired Ca²⁺ handling in the sarcoplasmic reticulum (SR), leading to reduced cardiac function. We have recently demonstrated that expression of mRNA encoding sarco(endo)plasmic reticulum Ca²⁺-ATPase 2 (SERCA2), a major Ca²⁺ transport protein in SR, is markedly decreased in DOX-treated hearts. To extend this observation, we have dissected the molecular mechanisms by which DOX downregulates SERCA2 gene transcription. Using cultured rat neonatal cardiac myocytes, we found that the antioxidant N-acetylcysteine blocked the DOX-induced decrease in SERCA2 mRNA levels, as well as the DOX-induced increase in H₂O₂ concentration; thus, H₂O₂ is an intracellular mediator of DOX activity. Using a luciferase reporter assay, we found that the sequence from -284 to -72 bp in the 5' flanking region of the SERCA2 gene has a DOX-responsive element. Although several transcription factors have putative binding motifs in this region of the SERCA2 gene, only the expression of Egr-1 mRNA and the binding of Egr-1 protein to the 5' regulatory sequence of SERCA2 gene increased markedly after DOX administration. We also found that overexpression of Egr-1 was associated with a significant reduction in SERCA2 gene transcription. In addition, Egr-1 antisense oligonucleotides blocked the DOX-induced reduction in SERCA2 mRNA, suggesting that Egr-1 is a transcriptional inhibitor of the SERCA2 gene in DOX-induced cardiomyopathy. We observed activation of 3 mitogen-activated protein kinases (MAPKs), p44/42 MAPK, p38 MAPK, and stress-activated MAPK/Jun N-terminal kinase, by DOX, but only a specific inhibitor of the p44/42 MAPK kinase suppressed the effects of DOX on Egr-1 and SERCA2 mRNA expression. These findings indicate that reactive oxygen intermediates, the transcription factor Egr-1, and p44/42 MAPK are critical elements in the transcriptional regulation of the SERCA2 gene in response to DOX.

Key Words: SERCA2 • doxorubicin • hydrogen peroxide • mitogen-activated protein kinase • Egr-1

This article has been cited by other articles:

				O. S. Bains, R. H. Takahashi, T. A. Pfeifer, T. A. Grigliatti, R. E. Reid, and K. W. Riggs Two Allelic Variants of Aldo-Keto Reductase 1A1 Exhibit Reduced in Vitro Metabolism of Daunorubicin Drug Metab. Dispos., May 1, 2008; 36(5): 904 - 910. [Abstract] [Full Text] [PDF]

				M. Shimura, S. Minamisawa, H. Takeshima, Q. Jiao, Y. Bai, S. Umemura, and Y. Ishikawa Sarcalumenin alleviates stress-induced cardiac dysfunction by improving Ca2+ handling of the sarcoplasmic reticulum Cardiovasc Res, January 15, 2008; 77(2): 362 - 370. [Abstract] [Full Text] [PDF]

				Y. Xu, C. Liu, J. C. Clark, and J. A. Whitsett Functional Genomic Responses to Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) and CFTR{Delta}508 in the Lung J. Biol. Chem., April 21, 2006; 281(16): 11279 - 11291. [Abstract] [Full Text] [PDF]

				Y.-C. Lien, T. Noel, H. Liu, A. J. Stromberg, K.-C. Chen, and D. K. St. Clair Phospholipase C-{delta}1 Is a Critical Target for Tumor Necrosis Factor Receptor-Mediated Protection against Adriamycin-Induced Cardiac Injury. Cancer Res., April 15, 2006; 66(8): 4329 - 4338. [Abstract] [Full Text] [PDF]

				Y.-C. Lien, S.-M. Lin, R. Nithipongvanitch, T. D. Oberley, T. Noel, Q. Zhao, C. Daosukho, and D. K. St. Clair Tumor necrosis factor receptor deficiency exacerbated Adriamycin-induced cardiomyocytes apoptosis: an insight into the Fas connection. Mol. Cancer Ther., February 1, 2006; 5(2): 261 - 269. [Abstract] [Full Text] [PDF]

				C. Poizat, P. L. Puri, Y. Bai, and L. Kedes Phosphorylation-Dependent Degradation of p300 by Doxorubicin-Activated p38 Mitogen-Activated Protein Kinase in Cardiac Cells Mol. Cell. Biol., April 1, 2005; 25(7): 2673 - 2687. [Abstract] [Full Text] [PDF]

				T. Takaseya, M. Ishimatsu, E. Tayama, A. Nishi, T. Akasu, and S. Aoyagi Mechanical unloading improves intracellular Ca2+ regulation in rats with doxorubicin-induced cardiomyopathy J. Am. Coll. Cardiol., December 7, 2004; 44(11): 2239 - 2246. [Abstract] [Full Text] [PDF]

				W.-h. Feng, J. I. Cohen, S. Fischer, L. Li, M. Sneller, R. Goldbach-Mansky, N. Raab-Traub, H.-J. Delecluse, and S. C. Kenney Reactivation of Latent Epstein-Barr Virus by Methotrexate: A Potential Contributor to Methotrexate-Associated Lymphomas J Natl Cancer Inst, November 17, 2004; 96(22): 1691 - 1702. [Abstract] [Full Text] [PDF]

				R. Vlasblom, A. Muller, R. J.P Musters, M. J Zuidwijk, C. van Hardeveld, W. J Paulus, and W. S Simonides Contractile arrest reveals calcium-dependent stimulation of SERCA2a mRNA expression in cultured ventricular cardiomyocytes Cardiovasc Res, August 15, 2004; 63(3): 537 - 544. [Abstract] [Full Text] [PDF]

				W.-h. Feng, G. Hong, H.-J. Delecluse, and S. C. Kenney Lytic Induction Therapy for Epstein-Barr Virus-Positive B-Cell Lymphomas J. Virol., February 15, 2004; 78(4): 1893 - 1902. [Abstract] [Full Text] [PDF]

				G. W. Small, S. Somasundaram, D. T. Moore, Y. Y. Shi, and R. Z. Orlowski Repression of Mitogen-Activated Protein Kinase (MAPK) Phosphatase-1 by Anthracyclines Contributes to Their Antiapoptotic Activation of p44/42-MAPK J. Pharmacol. Exp. Ther., December 1, 2003; 307(3): 861 - 869. [Abstract] [Full Text] [PDF]

				K. Niwano, M. Arai, K. Tomaru, T. Uchiyama, Y. Ohyama, and M. Kurabayashi Transcriptional Stimulation of the eNOS Gene by the Stable Prostacyclin Analogue Beraprost Is Mediated Through cAMP-Responsive Element in Vascular Endothelial Cells: Close Link Between PGI2 Signal and NO Pathways Circ. Res., September 19, 2003; 93(6): 523 - 530. [Abstract] [Full Text] [PDF]

				S. Heon, M. Bernier, N. Servant, S. Dostanic, C. Wang, G. M. Kirby, L. Alpert, and L. E. Chalifour Dexrazoxane does not protect against doxorubicin-induced damage in young rats Am J Physiol Heart Circ Physiol, July 11, 2003; 285(2): H499 - H506. [Abstract] [Full Text] [PDF]

				F. D. Ragione, V. Cucciolla, V. Criniti, S. Indaco, A. Borriello, and V. Zappia p21Cip1 Gene Expression Is Modulated by Egr1: A NOVEL REGULATORY MECHANISM INVOLVED IN THE RESVERATROL ANTIPROLIFERATIVE EFFECT J. Biol. Chem., June 20, 2003; 278(26): 23360 - 23368. [Abstract] [Full Text] [PDF]