Document Detail


Expression of insulin-like growth factor-binding protein 5 in neuroblastoma cells is regulated at the transcriptional level by c-Myb and B-Myb via direct and indirect mechanisms.
MedLine Citation:
PMID:  11973331     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Neuroblastoma (NB), a malignant childhood tumor deriving from the embryonic neural crest, is sensitive to the growth-stimulating effects of insulin-like growth factors (IGFs). Aggressive cases of this disease often acquire autocrine loops of IGF production, but the mechanisms through which the different components of the IGF axis are regulated in tumor cells remain unclear. Upon conditional expression of c-Myb in a NB cell line, we detected up-regulation of IGF1, IGF1 receptor, and insulin-like growth factor-binding protein 5 (IGFBP-5) expression. Analysis of the IGFBP-5 promoter revealed two potential Myb binding sites at position -59 to -54 (M1) and -429 to -424 (M2) from the transcription start site; both sites were bound by c-Myb and B-Myb in vitro and in vivo. Reporter assays carried out using the proximal region of the human IGFBP-5 promoter demonstrated that c-Myb and B-Myb enhanced transcription. However, site-directed mutagenesis and deletion of the Myb binding sites coupled with reporter assays revealed that M2 but not M1 was important for Myb-dependent transactivation of the IGFBP-5 promoter. The double mutant M1/M2 was still transactivated by c-Myb, suggesting the existence of Myb binding-independent mechanisms of IGFBP-5 promoter regulation. A constitutively active AKT transactivated the IGFBP-5 promoter, whereas the phosphatidylinositol 3-kinase inhibitor LY294002 suppressed it. Moreover, the kinase dead dominant negative K179M AKT mutant was able to inhibit transcription from the M2 and M1/M2 IGFBP-5 mutant promoters. Deletion analysis of the IGFBP-5 promoter revealed that the AKT-responsive region lies between nucleotides -334 and -83. Together, these data suggest that the Myb binding-independent transactivation of the IGFBP-5 promoter was due to the activation of the phosphatidylinositol 3-kinase/AKT pathway likely mediated by IGF1 receptor-dependent signals. Finally, IGFBP-5 was able to modulate proliferation of NB cells in a manner dependent on its concentration and on the presence of IGFs.
Authors:
Barbara Tanno; Anna Negroni; Roberta Vitali; Maria Celeste Pirozzoli; Vincenzo Cesi; Camillo Mancini; Bruno Calabretta; Giuseppe Raschellà
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.     Date:  2002-04-24
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  277     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2002 Jun 
Date Detail:
Created Date:  2002-06-24     Completed Date:  2002-08-06     Revised Date:  2012-06-22    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  23172-80     Citation Subset:  IM    
Affiliation:
Ente Nuove Tecnologie Energia Ambiente (ENEA), Section of Toxicology and Biomedical Sciences, Via Anguillarese 301, 00060 S. Maria di Galeria, Rome, Italy.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Cycle Proteins*
Cell Division
DNA-Binding Proteins / genetics*
Gene Expression Regulation, Neoplastic*
Genes, myb / physiology*
Humans
Insulin-Like Growth Factor Binding Protein 5 / genetics*
Insulin-Like Growth Factor II / physiology
Mice
Neuroblastoma / metabolism*
Promoter Regions, Genetic
Protein-Serine-Threonine Kinases*
Proto-Oncogene Proteins / physiology
Proto-Oncogene Proteins c-akt
Trans-Activators / genetics*
Transcription, Genetic*
Transcriptional Activation
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Cell Cycle Proteins; 0/DNA-Binding Proteins; 0/Insulin-Like Growth Factor Binding Protein 5; 0/MYBL2 protein, human; 0/Mybl2 protein, mouse; 0/Proto-Oncogene Proteins; 0/Trans-Activators; 67763-97-7/Insulin-Like Growth Factor II; EC 2.7.11.1/AKT1 protein, human; EC 2.7.11.1/Protein-Serine-Threonine Kinases; EC 2.7.11.1/Proto-Oncogene Proteins c-akt

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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