title: Visualization of oxidative processes at the cellular level during neutrophil-mediated cytotoxicity against a human hepatoma cell line, HCC-M.
abstract: Human neutrophil-mediated oxidative processes against a human hepatoma cell line, HCC-M, was visualized at the cellular level by using a silicon-intensified target camera and subsequently processing with a computer-assisted digital-imaging processor. Neutrophils were activated by a streptococcal preparation, OK-432. A hydroperoxide-sensitive tracer, dichlorofluorescein diacetate, was loaded in HCC-M and temporal and spatial changes of lipid peroxides in this cell after addition of stimulated neutrophils were analyzed. The luminol-dependent chemiluminescence activity of neutrophils was significantly enhanced and continued for at least 2 hr by stimulation with OK-432, and its activity was shown to be accumulated at the site where a neutrophil attached with HCC-M. The intensity of dichlorofluorescein fluorescence in HCC-M rapidly increased after adding stimulated neutrophils, and their reaction was significantly attenuated by superoxide dismutase. The number of non-viable cells was increased as the dichlorofluorescein fluorescence increase. It is suggested that oxidative stress may play an important role in neutrophil-mediated tumor-cell damage.
authors.affiliation: Department of Internal Medicine, School of Medicine, Keio University, Tokyo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Saito
authors.forename: D
authors.initials: D
authors.lastname: Fukumura
authors.forename: I
authors.initials: I
authors.lastname: Kurose
authors.forename: M
authors.initials: M
authors.lastname: Suematsu
authors.forename: S
authors.initials: S
authors.lastname: Tada
authors.forename: T
authors.initials: T
authors.lastname: Kagawa
authors.forename: S
authors.initials: S
authors.lastname: Miura
authors.forename: T
authors.initials: T
authors.lastname: Morizane
authors.forename: M
authors.initials: M
authors.lastname: Tsuchiya
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D005452:Fluoresceins / Q000494:pharmacology; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism*; D008163:Luminescent Measurements; D009504:Neutrophils / Q000378:metabolism*; D010084:Oxidation-Reduction / Q000187:drug effects; D010844:Picibanil / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: D005452:Fluoresceins; C029569:diacetyldichlorofluorescein; D010844:Picibanil; D013755:Tetradecanoylphorbol Acetate