Induction of myeloperoxidase and nitrotyrosine formation in a human eosinophilic leukemia cell line, EoL-1.
The human eosinophilic leukemia cell line, EoL-1, differentiated with butyrate as an eosinophilic cellular model was evaluated for peroxidase-dependent tyrosine nitration. Butyrate suppressed cell growth and induced eosinophilic granules in EoL-1 cells after 9 days of culture. Peroxidase activity was detected biochemically and histochemically from 3-day cultures and it increased in a time dependent manner. This peroxidase activity was inhibited by cyanide. Nitrotyrosine formation catalysed by peroxidase using hydrogen peroxide and nitrite was detected at a high level similar to that of mature eosinophils. However, no expression of eosinophil peroxidase (EPO) was detected by RT-PCR or immunocytochemistry. In contrast, the induction of myeloperoxidase (MPO) by butyrate was clearly detected by RT-PCR, Northern blot, and immunocytochemical staining. These results suggest that butyrate induces MPO rather than EPO in EoL-1 cells and that the formation of nitrotyrosine in butyrate-induced cells is dependent on MPO.
Kodama
Norio
N
Department of Environmental and Preventive Medicine, Graduate School of Medical Science, Kanazawa University, 13-1 Takara-machi, 920-8640 Kanazawa, Japan.
Kambayashi
Yasuhiro
Y
Kubo
Masayuki
M
Nobukuni
Yoshitaka
Y
Kimura
Shingo
S
Nakamura
Hiroyuki
H
Ogino
Keiki
K
D002087:Butyrates / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D003594:Cytoplasmic Granules / Q000187:drug effects; D047209:Eosinophil Peroxidase; D006801:Humans; D017681:Hypereosinophilic Syndrome / Q000188:drug therapy / Q000201:enzymology* / Q000378:metabolism; D007150:Immunohistochemistry; D009195:Peroxidase / Q000378:metabolism*; D010544:Peroxidases / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000096:biosynthesis*
D002087:Butyrates; C002744:3-nitrotyrosine; D014443:Tyrosine; D047209:Eosinophil Peroxidase; D010544:Peroxidases; D009195:Peroxidase