title: Manganese superoxide dismutase overexpression changes plating efficiency bidirectionally according to change in redox for SaOS2 human osteosarcoma cell line.
abstract: Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that converts cytotoxic superoxide radicals into hydrogen peroxide. MnSOD activity is lower in tumor cells, and MnSOD overexpression reportedly ameliorates malignant phenotypes. We established stable MnSOD overexpressing cell lines from a human osteosarcoma cell line, SaOS2, and then investigated the effects of MnSOD overexpression on plating efficiency (PE) and the involvement of reactive oxygen species, including nitric oxide (NO) in those effects. The PE of SaOS2FM(L), a moderate MnSOD overexpression cell line, increased, while that of SaOS2FM(H), a high MnSOD overexpression cell line, decreased. Although we assessed PE using a colony-formation assay, time-lapse microscopic observation revealed that cells attached to the flasks had undergone neither apoptosis nor necrosis. Moreover, MnSOD overexpression did not affect cell doubling time. Therefore, MnSOD overexpression might correlate directly with cellular adhesion's effect on PE changes. When L-buthionine-[S,R]-sulfoximine (BSO) was administered to increase the intracellular concentration of hydrogen peroxide, the PEs of both cell lines decreased, and when hydrogen peroxide was eliminated by the administration of sodium pyruvate, only the PE of SaOS2FM(H) increased. The combination of BSO and NO (NOR4 or isosorbide 5-mononitrate) administration synergistically decreased PE in both cell lines. These findings suggest that changes in cellular adhesion properties correlate with the balance between increased hydrogen peroxide levels and decreased superoxide radical levels. This is the first report to indicate that PE and cellular adhesion properties change bidirectionally according to the levels of MnSOD overexpression: first increasing then decreasing as MnSOD activity increases. Our results indicate that PE changes might be decided by the balance between two cytotoxic compounds (decreased superoxide radical levels and increased hydrogen peroxide levels), and that NO loading and increased hydrogen peroxide synergistically reduce PE and cellular adhesion.
authors.affiliation: Department of Radiology, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
authors.forename: Megumi
authors.initials: M
authors.lastname: Komatsu
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kuroda
authors.forename: Yadi
authors.initials: Y
authors.lastname: Wang
authors.forename: Daret
authors.initials: D
authors.lastname: St Clair
authors.forename: Muneyasu
authors.initials: M
authors.lastname: Urano
authors.forename: Shiro
authors.initials: S
authors.lastname: Akaki
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Asaumi
authors.forename: Shoji
authors.initials: S
authors.lastname: Kawasaki
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Hiraki
authors.forename: Susumu
authors.initials: S
authors.lastname: Kanazawa
mesh_terms: D001859:Bone Neoplasms / Q000201:enzymology* / Q000473:pathology; D002448:Cell Adhesion / Q000187:drug effects*; D003114:Colony-Forming Units Assay; D015972:Gene Expression Regulation, Neoplastic; D016158:Genes, p53; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D009569:Nitric Oxide / Q000494:pharmacology; D012516:Osteosarcoma / Q000201:enzymology* / Q000473:pathology*; D016877:Oxidants / Q000737:chemistry; D010641:Phenotype; D017382:Reactive Oxygen Species / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000096:biosynthesis* / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D015854:Up-Regulation
chemical_list: D016877:Oxidants; D017382:Reactive Oxygen Species; D009569:Nitric Oxide; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase