title: Hypoxia-mediated alterations and their role in the HER-2/neuregulated CREB status and localization.
abstract: The cAMP-responsive element-binding protein (CREB) is involved in the tumorigenicity of HER-2/neu-overexpressing murine and human tumor cells, but a link between the HER-2/neu-mediated CREB activation, its posttranslational modification and localization and changes in the cellular metabolism, due to an altered (tumor) microenvironment remains to be established. The present study demonstrated that shRNA-mediated silencing of CREB in HER-2/neu-transformed cells resulted in decreased tumor formation, which was associated with reduced angiogenesis, but increased necrotic and hypoxic areas in the tumor. Hypoxia induced pCREBSer133, but not pCREBSer121 expression in HER-2/neu-transformed cells. This was accompanied by upregulation of the hypoxia-inducible genes GLUT1 and VEGF, increased cell migration and matrix metalloproteinase-mediated invasion. Treatment of HER-2/neu+ cells with signal transduction inhibitors targeting in particular HER-2/neu was able to revert hypoxia-controlled CREB activation. In addition to changes in the phosphorylation, hypoxic response of HER-2/neu+ cells caused a transient ubiquitination and SUMOylation as well as a co-localization of nuclear CREB to the mitochondrial matrix. A mitochondrial localization of CREB was also demonstrated in hypoxic areas of HER-2/neu+ mammary carcinoma lesions. This was accompanied by an altered gene expression pattern, activity and metabolism of mitochondria leading to an increased respiratory rate, oxidative phosphorylation and mitochondrial membrane potential and consequently to an enhanced apoptosis and reduced cell viability. These data suggest that the HER-2/neu-mediated CREB activation caused by a hypoxic tumor microenvironment contributes to the neoplastic phenotype of HER-2/neu+ cells at various levels.
authors.affiliation: Institute of Medical Immunology, Martin Luther University Halle-Wittenberg, Halle, Germany.
authors.forename: André
authors.initials: A
authors.lastname: Steven
authors.affiliation: Institute of Medical Immunology, Martin Luther University Halle-Wittenberg, Halle, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Leisz
authors.affiliation: Charité, Institute of Pathology, Berlin, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Sychra
authors.affiliation: Centre for Basic Medical Research, Martin Luther University Halle-Wittenberg, Halle, Germany.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Hiebl
authors.affiliation: Institute of Pathology, Martin Luther University Halle-Wittenberg, Halle, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Wickenhauser
authors.affiliation: Department of Internal Medicine 5, Hematology and Oncology, University of Erlangen-Nuremberg, Erlangen, Germany.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Mougiakakos
authors.affiliation: Karolinska Institutet, CCK, Stockholm, Sweden.
authors.forename: Rolf
authors.initials: R
authors.lastname: Kiessling
authors.affiliation: Charité, Institute of Pathology, Berlin, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Denkert
authors.affiliation: Institute of Medical Immunology, Martin Luther University Halle-Wittenberg, Halle, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Seliger
mesh_terms: D000818:Animals; D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000378:metabolism* / Q000473:pathology; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D006801:Humans; D008325:Mammary Neoplasms, Experimental / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism*; D059016:Tumor Microenvironment / Q000502:physiology
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D018719:Erb-b2 Receptor Tyrosine Kinases
keywords: CREB; HER-2/neu; angiogenesis; hypoxia; mitochondria