title: Hypoxia effects on proangiogenic factors in human umbilical vein endothelial cells: functional role of the peptide somatostatin.
abstract: The aim of this study was to investigate hypoxia effects on vascular endothelial growth factor (VEGF) and its receptors VEGFR-1 and VEGFR-2 in human umbilical vein endothelial cells (HUVEC) and to determine their modulation by the peptide somatostatin (SRIF) and its analogues. The involvement of signal transducer and activator of transcription (STAT) 3 and hypoxia inducible factor (HIF)-1 was also investigated. Quantitative real-time PCR, Western blot and ELISA were used. Hypoxia upregulated VEGF expression and release, whereas it downregulated VEGFR-1 and VEGFR-2. In contrast, neither the expression nor the phosphorylation of the platelet-derived growth factor receptor (PDGFR) β was affected by hypoxia. SU1498 at 1 μM did not affect pVEGFR-2 and pPDGFRβ, whereas at 20 μM it inhibited pVEGFR-2, but not pPDGFRβ. Upregulated VEGF expression and release were prevented by SU1498, which also inhibited the hypoxia-induced pSTAT3 and HIF-1α. Blocking pSTAT3 with S3I-201 inhibited HIF-1α and VEGF upregulation, suggesting the existence of an autocrine loop involving STAT3, HIF-1, VEGF and VEGFR-2. Endothelial cells express somatostatin (SRIF) receptors (sst(1-5)) although less is known in HUVEC. We found that sst(1) and sst(4) were expressed by HUVEC with sst(1) more expressed than sst(4) mRNA. Hypoxia downregulated sst(1), whereas it upregulated sst(4). The sst(1) downregulation, but not the sst(4) upregulation, was prevented by SU1498, S3I-201 or YC-1, an inhibitor of HIF-1α. SRIF and the sst(1) agonist CH-275, but not the sst(4) agonist L803,087 and the sst(2)/sst(3)/sst(5) agonist octreotide, prevented hypoxia effects on VEGF and its receptors. In addition, SRIF and CH-275 inhibited the hypoxia-induced pSTAT3 and HIF-1α accumulation. Our results suggest that SRIF acting at sst(1) limits upregulated VEGF expression and release through a control on the activity of STAT3 and HIF-1, supporting the possible use of sst(1) agonists in antiangiogenic therapies.
authors.affiliation: Department of Biology, University of Pisa, via San Zeno, 31, 56127 Pisa, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Dal Monte
authors.forename: Davide
authors.initials: D
authors.lastname: Martini
authors.forename: Chiara
authors.initials: C
authors.lastname: Ristori
authors.forename: Danilo
authors.initials: D
authors.lastname: Azara
authors.forename: Chiara
authors.initials: C
authors.lastname: Armani
authors.forename: Alberto
authors.initials: A
authors.lastname: Balbarini
authors.forename: Paola
authors.initials: P
authors.lastname: Bagnoli
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D015153:Blotting, Western; D015687:Cell Hypoxia; D015536:Down-Regulation; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D009389:Neovascularization, Pathologic / Q000188:drug therapy; D016133:Polymerase Chain Reaction; D017481:Receptors, Somatostatin / Q000235:genetics / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism; D013004:Somatostatin / Q000031:analogs & derivatives / Q000494:pharmacology*; D014471:Umbilical Veins / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics / Q000378:metabolism*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism*
chemical_list: D020533:Angiogenesis Inhibitors; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D017481:Receptors, Somatostatin; D050796:STAT3 Transcription Factor; D042461:Vascular Endothelial Growth Factor A; D013004:Somatostatin; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2