title: Matrix metalloproteinase-2 and -9 are induced differently by doxorubicin in H9c2 cells: The role of MAP kinases and NAD(P)H oxidase.
abstract: OBJECTIVE
  Dysregulation of myocardial metalloproteinases (MMPs) is now regarded as an early contributory mechanism for the initiation and progression of heart failure. Doxorubicin is a strongly cardiotoxic anticancer drug. This study investigates the effects of doxorubicin on myocardial MMP-2 and MMP-9 activation.
  
  
  METHODS
  After pre-treatment with or without carvedilol or dexrazoxane, we exposed H9c2 cardiomyocytes to doxorubicin to evaluate reactive oxygen species (ROS) formation and MMP-2 and MMP-9 expression and activation. To investigate the signaling pathways leading to doxorubicin-induced MMP activation, we also examined the phosphorylation of three members of the MAPK family (ERK1/2, p38, and JNK), the effects of selective inhibitors of ERK1/2, p38, and JNK on MMP transcription and activity, the transcription of the NAD(P)H oxidase subunit Nox1, and the effects of the NAD(P)H oxidase inhibitor DPI on MMP activation.
  
  
  RESULTS
  Doxorubicin induces a significant increase in ROS formation and a rapid increase of MMP expression and activation. Pre-treatment with carvedilol or dexrazoxane prevented these effects. We also found that p38 is the MAPK that is mainly responsible for MMP-9 activation through an NAD(P)H-independent mechanism. ERK and JNK modulate the transcription of the NAD(P)H oxidase subunit Nox1, while the JNK/ERK NAD(P)H oxidase cascade is an important pathway that mediates doxorubicin signaling to MMP-2. Inhibition of NAD(P)H oxidase attenuates the increase in MMP-2, but augments the doxorubicin-induced increase in MMP-9.
  
  
  CONCLUSIONS
  Enhancement of MMP-2 and MMP-9 in cardiac myocytes in response to doxorubicin is mediated by the cooperation of ERK, JNK, and p38 kinase pathways, most of which are redox dependent.
authors.affiliation: Research Center of Cardiovascular Biology, Department of Cardiology, University of Genova, Viale Benedetto XV, 6 16132 Genova, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Spallarossa
authors.forename: Paola
authors.initials: P
authors.lastname: Altieri
authors.forename: Silvano
authors.initials: S
authors.lastname: Garibaldi
authors.forename: Giorgio
authors.initials: G
authors.lastname: Ghigliotti
authors.forename: Chiara
authors.initials: C
authors.lastname: Barisione
authors.forename: Valeria
authors.initials: V
authors.lastname: Manca
authors.forename: Patrizia
authors.initials: P
authors.lastname: Fabbi
authors.forename: Alberto
authors.initials: A
authors.lastname: Ballestrero
authors.forename: Claudio
authors.initials: C
authors.lastname: Brunelli
authors.forename: Antonio
authors.initials: A
authors.lastname: Barsotti
mesh_terms: D000873:Anthracenes / Q000494:pharmacology; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D015153:Blotting, Western; D002227:Carbazoles / Q000494:pharmacology; D000077261:Carvedilol; D002395:Catecholamines / Q000494:pharmacology; D002460:Cell Line; D004317:Doxorubicin / Q000494:pharmacology*; D004789:Enzyme Activation; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D048288:Imidazolines / Q000494:pharmacology; D053613:Janus Kinase 1; D020935:MAP Kinase Signaling System; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000201:enzymology*; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000502:physiology*; D011412:Propanolamines / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D011725:Pyridines / Q000494:pharmacology; D011929:Razoxane / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors
chemical_list: D000873:Anthracenes; D000903:Antibiotics, Antineoplastic; D000975:Antioxidants; D002227:Carbazoles; D002395:Catecholamines; D005419:Flavonoids; D007093:Imidazoles; D048288:Imidazolines; D011412:Propanolamines; D011725:Pyridines; D000077261:Carvedilol; C432165:pyrazolanthrone; D011929:Razoxane; D004317:Doxorubicin; C011775:(3,4-dihydroxyphenylamino)-2-imidazoline; D019255:NADPH Oxidases; D011505:Protein-Tyrosine Kinases; C507904:JAK1 protein, human; D053613:Janus Kinase 1; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D020782:Matrix Metalloproteinases; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; C093642:SB 203580; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one