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Thromb Haemost 2005; 93(06): 1176-1184
DOI: 10.1160/TH04-11-0761
Cell Signalling and Vessel Remodelling
Schattauer GmbH

Transcriptional regulation of plasminogen activator inhibitor-1 expression by insulin-like growth factor-1 via MAP kinases and hypoxia-inducible factor-1 in HepG2 cells

Elitsa Y. Dimova
2   Department Chemistry/Biochemistry, University of Kaiserslautern, Kaiserslautern, Germany
1   Institute of Biochemistry and Molecular Cell Biology, Göttingen, Germany
,
Ulrike Möller
1   Institute of Biochemistry and Molecular Cell Biology, Göttingen, Germany
,
Stephan Herzig
3   Department of Molecular Metabolic Control (A170), German Cancer Research Center, Heidelberg, Heidelberg, Germany
,
Trine Fink
4   Laboratory for Stem Cell Research, Aarhus C, Denmark
,
Vladimir Zachar
4   Laboratory for Stem Cell Research, Aarhus C, Denmark
,
Peter Ebbesen
4   Laboratory for Stem Cell Research, Aarhus C, Denmark
,
Thomas Kietzmann
2   Department Chemistry/Biochemistry, University of Kaiserslautern, Kaiserslautern, Germany
1   Institute of Biochemistry and Molecular Cell Biology, Göttingen, Germany
› Author AffiliationsFinancial support: This study was supported by the Deutsche Forschungsgemeinschaft SFB 402 Teilprojekt A1, GRK 335 and Fonds der Chemischen Industrie
Further Information

Publication History

Received 24 November 2004

Accepted after resubmission 17 March 2005

Publication Date:
11 December 2017 (online)

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Summary

Insulin-like growth factor 1 (IGF-1) and plasminogen activator inhibitor-1 (PAI-1) appear to play a crucial role in a number of processes associated with growth and tissue remodelling. IGF-1 was shown to enhance PAI-1 expression in primary hepatocytes and HepG2 hepatoma cells, but the molecular mechanisms underlying this effect have not been fully elucidated. In this study, we investigated the transcriptional mechanism and the signaling pathway by which IGF-1 mediates induction of PAI-1 expression in HepG2 cells. By using human PAI-1 promoter reporter gene assays we found that mutation of the hypoxia responsive element (HRE), which could bind hypoxia-inducible factor-1 (HIF-1), nearly abolished the induction by IGF-1. We found that IGF-1-induced up-regulation of PAI-1 expression was associated with activation of HIF-1α. Furthermore, IGF-1 enhanced HIF-1α protein levels and HIF-1 DNA-binding to each HRE, E4 and E5 as shown by EMSA. Mutation of the E-boxes, E4 and E5, did not affect the IGF-1-dependent induction of PAI-1 promoter constructs under normoxia but abolished the effect of IGF-1 under hypoxia. Inhibition of either the PI3K by LY294002 or ERK1/2 by U0126 reduced HIF-1α protein levels while both inhibitors together completely abolished the IGF-1 effect on HIF-1α. Remarkably, transfection of HepG2 cells with vectors expressing a dominant-negative PDK1 or the PKB inhibitor, TRB3, did not influence while dominant-negative Raf inhibited the IGF-1 effect on HIF-1α. Thus, IGF-1 activates human PAI-1 gene expression through activation of the PI3-kinase and ERK1/2 via HIF-1α.

Keywords

Plasminogen activator inhibitor-1 - insulin-like growth factor - hypoxia-inducible factor-1 - phosphatidylinositol-dependent kinase-1 - extracellular regulated kinase
 

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