title: Necrotic pathway in human osteosarcoma Saos-2 cell death induced by chloroacetaldehyde.
abstract: Chloroacetaldehyde, a metabolite of the anticancer drug ifosfamide, may be responsible for serious adverse effects like encephalopathy in ifosfamide chemotherapy. In this study, we demonstrate that chloroacetaldehyde, but not ifosfamide, induces cell death in human osteosarcoma Saos-2 cells and we investigated the mechanism by which this occurs. Chloroacetaldehyde above 30 micromol/l induced significant cell death in a time-dependent manner. Thiol compounds such as N-acetyl cysteine, glutathione and dithiothreitol protected the cells against chloroacetaldehyde-induced cell death, although other nonthiol compounds and the antioxidative enzymes superoxide dismutase and catalase did not, suggesting that reactive oxygen species might not mediate cell death. In cells exposed to chloroacetaldehyde, levels of both total thiols and glutathione were significantly reduced. Chloroacetaldehyde also collapsed the mitochondrial membrane potential of these cells, induced the release of cytochrome c from mitochondria to the cytosol and significantly reduced cellular ATP levels during the course of death. The mitochondrial potential collapse was also prevented by thiol compounds. Flow cytometric analyses by means of annexin-V and propidium iodide double staining and immunofluorescence staining of active caspase-3 revealed that cells subjected to a lethal dose of chloroacetaldehyde displayed features characteristic of necrosis and that caspase-3 was not activated in response to chloroacetaldehyde. Taken together, these findings suggest that Saos-2 cells exposed to chloroacetaldehyde die by necrosis resulting from a decrease in intracellular thiols, disruption of the mitochondrial membrane potential and the depletion of cellular ATP.
authors.affiliation: Department of Biochemistry, Hokkaido Pharmaceutical University, Otaru, and National Hospital Organization Hokkaido Cancer Center, Sapporo, Hokkaido, Japan.
authors.forename: Kyohei
authors.initials: K
authors.lastname: Takahashi
authors.forename: Koichi
authors.initials: K
authors.lastname: Sakurai
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Takahashi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tanaka
authors.forename: Yukio
authors.initials: Y
authors.lastname: Fujimoto
mesh_terms: D000079:Acetaldehyde / Q000031:analogs & derivatives* / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000502:physiology; D017304:Annexin A5 / Q000378:metabolism; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology; D015153:Blotting, Western; D001859:Bone Neoplasms / Q000188:drug therapy* / Q000473:pathology; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005434:Flow Cytometry; D006801:Humans; D007069:Ifosfamide / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D051336:Mitochondrial Membranes / Q000187:drug effects; D009336:Necrosis; D012516:Osteosarcoma / Q000188:drug therapy* / Q000473:pathology; D011419:Propidium / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism
chemical_list: D017304:Annexin A5; D018906:Antineoplastic Agents, Alkylating; D013438:Sulfhydryl Compounds; D011419:Propidium; D000255:Adenosine Triphosphate; C004656:chloroacetaldehyde; D000079:Acetaldehyde; D007069:Ifosfamide