Visualization of oxidative processes at the cellular level during neutrophil-mediated cytotoxicity against a human hepatoma cell line, HCC-M.
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.
Saito
H
H
Department of Internal Medicine, School of Medicine, Keio University, Tokyo, Japan.
Fukumura
D
D
Kurose
I
I
Suematsu
M
M
Tada
S
S
Kagawa
T
T
Miura
S
S
Morizane
T
T
Tsuchiya
M
M
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
D005452:Fluoresceins; C029569:diacetyldichlorofluorescein; D010844:Picibanil; D013755:Tetradecanoylphorbol Acetate