title: Interleukin-1β induced nuclear factor-κB binds to a disintegrin-like and metalloproteinase with thrombospondin type 1 motif 9 promoter in human chondrosarcoma cells.
abstract: Nuclear factor-κB (NF-κB) is involved in the regulation of inflammation‑associated genes. NF-κB forms dimers which bind with sequences referred to as NF-κB sites (9-11 bp). A disintegrin-like and metalloproteinase with thrombospondin type 1 motif 9 (ADAMTS9) is a type of proteoglycanase, which proteolytically cleaves versican and aggrecan. ADAMTS9 is a cytokine-inducible gene that contains binding sites for NF-κB within its promoter region. Interleukin-1β (IL-1β) affects cartilage metabolism and is involved in the NF-κB pathway. It is therefore hypothesized that NF-κB binding with ADAMTS9 promoters may activate IL-1β, thereby promoting chondrocytic cell growth. In the present study, the OUMS-27 chondrocytic human chondrosarcoma cell line was treated with IL-1β with or without inhibitors of NF-κB signaling pathways. Chromatin immunoprecipitation (ChIP) and electromobility shift assays (EMSA) were conducted order to analyze the binding of NF-κB with the ADAMTS9 promoter region. NF-κB-p65 subunit phosphorylation was promoted in IL-1β-treated cells, which were not treated with inhibitors of NF-κB signaling pathways. By contrast, NF-κB-p65 subunit phosphorylation was inhibited in cells that had been treated with BAY-117085, an NF-κB pathway inhibitor. ChIP and EMSA assays demonstrated that, following treatment with IL-1β, NF-κB‑p65 bound to elements located at -1177 and -1335 in the ADAMTS9 promoter region, in contrast to the untreated samples. The results of the present study suggested that NF-κB may be involved in IL-1β-induced activation of ADAMTS9 in human chondrocytes.
authors.affiliation: Division of Chemistry, Ankara Regional Office of Council of Forensic Medicine, Ankara 06100, Turkey.
authors.forename: Aynur
authors.initials: A
authors.lastname: Altuntas
authors.affiliation: Pediatric Immunology Unit, Institute of Children's Health, Hacettepe University, Ankara 06100, Turkey.
authors.forename: Sevil Oskay
authors.initials: SO
authors.lastname: Halacli
authors.affiliation: Faculty of Education, Division of Biology Education, Gazi University, Ankara 06500, Turkey.
authors.forename: Ozlem
authors.initials: O
authors.lastname: Cakmak
authors.affiliation: Department of Clinical Biochemistry, Medical Faculty, Hacettepe University, Ankara 06100, Turkey.
authors.forename: Gonul
authors.initials: G
authors.lastname: Erden
authors.affiliation: Department of Medical Biology, Turgut Ozal University School of Medicine, Ankara 06105, Turkey.
authors.forename: Sumeyya
authors.initials: S
authors.lastname: Akyol
authors.affiliation: Department of Medical Biochemistry, Dumlupinar University Medical Faculty, Kutahya 43100, Turkey.
authors.forename: Veli
authors.initials: V
authors.lastname: Ugurcu
authors.affiliation: Okayama University Graduate School of Medicine, Dentistry and Pharmacological Sciences, Okayama 700‑8558, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Hirohata
authors.affiliation: Department of Medical Biology, Turgut Ozal University School of Medicine, Ankara 06105, Turkey.
authors.forename: Kadir
authors.initials: K
authors.lastname: Demircan
mesh_terms: D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism*; D000071119:ADAMTS9 Protein; D001859:Bone Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D019902:Chondrocytes / Q000378:metabolism / Q000473:pathology; D002813:Chondrosarcoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D053583:Interleukin-1beta / Q000008:administration & dosage / Q000378:metabolism*; D009570:Nitriles / Q000008:administration & dosage; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D013450:Sulfones / Q000008:administration & dosage; D051996:Transcription Factor RelA / Q000235:genetics / Q000378:metabolism*
chemical_list: C416282:BAY 11-7085; D053583:Interleukin-1beta; D009570:Nitriles; D013450:Sulfones; D051996:Transcription Factor RelA; D051722:ADAM Proteins; D000071119:ADAMTS9 Protein; C412869:ADAMTS9 protein, human