title: On the role of 25-hydroxycholesterol synthesis by glioblastoma cell lines. Implications for chemotactic monocyte recruitment.
abstract: Glioblastoma multiforme (GBM) is the most common malignant primary brain tumor and is invariably fatal to affected patients. Oxysterols belong to a class of bioactive lipids that are implicated in neurological disease and are associated with various types of cancer. Here, we investigated expression and transcriptional regulation of cholesterol 25-hydroxylase (CH25H) in human U87MG and GM133 glioblastoma cell lines. We demonstrate that in both cell lines transcription and translation of CH25H are increased in response to TNFα and IL1β. In parallel, both cell lines upregulate 25-hydroxycholesterol (25-OHC) synthesis and secretion to levels comparable to bone marrow-derived mouse macrophages under inflammatory conditions. To determine whether 25-OHC acts as chemoattractant for tumor-associated macrophages, the human THP-1 monoblastic leukemia cell line was treated with varying amounts of the oxysterol. Experiments revealed that 25-OHC and lipid extracts isolated from GM133-conditioned medium (containing 7-fold higher 25-OHC concentrations than U87MG medium) induce chemotactic migration of THP-1 cells. Of note, 25-OHC also induced the migration of primary human peripheral blood monocytes. In response to exogenously added 25-OHC, THP-1 cells reorganized intermediate filament-associated vimentin to more cortical and polarized structures. Chemotactic migration of monocytes in response to 25-OHC was pertussis toxin-sensitive, indicating the involvement of G protein-coupled receptors. Using RNA interference we demonstrated that G protein-coupled receptor 183 (EBI2) contributes to 25-OHC-mediated chemotactic migration of THP-1 cells. These in vitro data indicate that GBM-derived and secreted 25-OHC may be involved in the recruitment of immune-competent cells to a tumor via EBI2.
authors.affiliation: Institute of Molecular Biology and Biochemistry, Medical University of Graz, Harrachgasse 21, Graz 8010, Austria.
authors.forename: Gerald
authors.initials: G
authors.lastname: Eibinger
authors.affiliation: Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Graz 8010, Austria.
authors.forename: Günter
authors.initials: G
authors.lastname: Fauler
authors.affiliation: Institute of Molecular Biology and Biochemistry, Medical University of Graz, Harrachgasse 21, Graz 8010, Austria.
authors.forename: Eva
authors.initials: E
authors.lastname: Bernhart
authors.affiliation: Institute of Molecular Biology and Biochemistry, Medical University of Graz, Harrachgasse 21, Graz 8010, Austria.
authors.forename: Sasa
authors.initials: S
authors.lastname: Frank
authors.affiliation: Institute of Cell Biology, Histology and Embryology, Medical University of Graz, Graz 8010, Austria.
authors.forename: Astrid
authors.initials: A
authors.lastname: Hammer
authors.affiliation: Institute of Molecular Biology and Biochemistry, Medical University of Graz, Harrachgasse 21, Graz 8010, Austria.
authors.forename: Andrea
authors.initials: A
authors.lastname: Wintersperger
authors.affiliation: Department of Neurosurgery, Medical University of Graz, Graz 8010, Austria.
authors.forename: Hans
authors.initials: H
authors.lastname: Eder
authors.affiliation: Institute of Experimental and Clinical Pharmacology, Medical University of Graz, Graz 8010, Austria.
authors.forename: Akos
authors.initials: A
authors.lastname: Heinemann
authors.affiliation: Ludwig Institute for Cancer Research, La Jolla, California, CA 92093, USA.
authors.forename: Paul S
authors.initials: PS
authors.lastname: Mischel
authors.affiliation: Institute of Molecular Biology and Biochemistry, Medical University of Graz, Harrachgasse 21, Graz 8010, Austria.
authors.forename: Ernst
authors.initials: E
authors.lastname: Malle
authors.affiliation: Institute of Molecular Biology and Biochemistry, Medical University of Graz, Harrachgasse 21, Graz 8010, Austria. Electronic address: wolfgang.sattler@medunigraz.at.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Sattler
mesh_terms: D001932:Brain Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D002633:Chemotaxis / Q000187:drug effects*; D005909:Glioblastoma / Q000378:metabolism*; D006801:Humans; D006888:Hydroxycholesterols / Q000378:metabolism*; D009000:Monocytes / Q000502:physiology*; D014176:Protein Biosynthesis; D034741:RNA, Small Interfering; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D013250:Steroid Hydroxylases / Q000235:genetics / Q000378:metabolism; D013261:Sterols / Q000494:pharmacology; D014158:Transcription, Genetic; D015854:Up-Regulation; D014746:Vimentin / Q000378:metabolism
chemical_list: C080105:GPR183 protein, human; D006888:Hydroxycholesterols; D034741:RNA, Small Interfering; D043562:Receptors, G-Protein-Coupled; D013261:Sterols; D014746:Vimentin; C007997:25-hydroxycholesterol; D013250:Steroid Hydroxylases; C507384:cholesterol 25-hydroxylase
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