MAPK pathway contributes to density- and hypoxia-induced expression of the tumor-associated carbonic anhydrase IX

Juraj Kopacek; Monika Barathova; Franck Dequiedt; Janka Sepelakova; Richard Kettmann; Jaromir Pastorek; Silvia Pastorekova

doi:10.1016/j.bbaexp.2005.03.003

MAPK pathway contributes to density- and hypoxia-induced expression of the tumor-associated carbonic anhydrase IX

Biochim Biophys Acta. 2005 May 25;1729(1):41-9. doi: 10.1016/j.bbaexp.2005.03.003. Epub 2005 Mar 23.

Authors

Juraj Kopacek¹, Monika Barathova, Franck Dequiedt, Janka Sepelakova, Richard Kettmann, Jaromir Pastorek, Silvia Pastorekova

Affiliation

¹ Centre of Molecular Medicine, Institute of Virology, Slovak Academy of Sciences, Dubravska cesta 9, 845 05 Bratislava, Slovakia.

PMID: 15833446
DOI: 10.1016/j.bbaexp.2005.03.003

Abstract

Transcription of the CA9 gene coding for a tumor-associated carbonic anhydrase IX (CA IX) isoform is regulated by hypoxia via the hypoxia-inducible factor 1 (HIF-1) and by high cell density via the phosphatidylinositol-3-kinase (PI3K) pathway. We examined the role of the mitogen-activated protein kinase (MAPK) pathway in the control of CA9 gene expression. Inhibition of MAPK signaling by U0126 in HeLa cells led to reduced activity of the PR1-HRE-luc CA9 promoter construct and decreased CA IX protein levels in dense culture as well as in hypoxia. Similar reduction was obtained by expression of a dominant-negative ERK1 mutant and was also observed in U0126-treated HIF-1alpha-deficient Ka13 cells. Simultaneous treatment with the MAPK and PI3K inhibitors U0126 and LY 294002 had stronger effect than individual inhibition of these pathways. Taken together, our results suggest that besides the PI3K pathway, the MAPK cascade is involved in the regulation of CA9 gene expression under both hypoxia and high cell density.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antigens, Neoplasm / biosynthesis*
Antigens, Neoplasm / genetics
Butadienes / pharmacology
Carbonic Anhydrase IX
Carbonic Anhydrases / biosynthesis*
Carbonic Anhydrases / genetics
Cell Count
Cell Hypoxia / drug effects
Cell Hypoxia / genetics
Cell Hypoxia / physiology
Chromones / pharmacology
DNA-Binding Proteins / metabolism
Enzyme Inhibitors / pharmacology
Gene Expression Regulation / drug effects
Gene Expression Regulation / physiology*
HeLa Cells
Humans
Hypoxia-Inducible Factor 1
Hypoxia-Inducible Factor 1, alpha Subunit
MAP Kinase Signaling System / drug effects
MAP Kinase Signaling System / physiology*
Mitogen-Activated Protein Kinase 3 / antagonists & inhibitors
Mitogen-Activated Protein Kinase 3 / genetics
Mitogen-Activated Protein Kinase 3 / metabolism
Morpholines / pharmacology
Nitriles / pharmacology
Nuclear Proteins / metabolism
Phosphatidylinositol 3-Kinases / metabolism
Phosphoinositide-3 Kinase Inhibitors
Transcription Factors / metabolism

Substances

Antigens, Neoplasm
Butadienes
Chromones
DNA-Binding Proteins
Enzyme Inhibitors
HIF1A protein, human
Hypoxia-Inducible Factor 1
Hypoxia-Inducible Factor 1, alpha Subunit
Morpholines
Nitriles
Nuclear Proteins
Phosphoinositide-3 Kinase Inhibitors
Transcription Factors
U 0126
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Mitogen-Activated Protein Kinase 3
CA9 protein, human
Carbonic Anhydrase IX
Carbonic Anhydrases