Functional assay of NF-kappaB translocation into nuclei by laser scanning cytometry: inhibitory effect by dexamethasone or theophylline.
Nuclear factor-kappa B (NF-kappaB)/Rel transcription factors play an important role in the inducible regulation of a variety of cytokine genes in epithelial cells. We assessed accumulation of fluorescence-stained NF-kappaB into propidium iodide-stained nuclei using laser scanning cytometry. "Activity-specific" antibodies to the Rel A (p65) and NF-kappaB1 (p50) subunits of NF-kappaB were detected in the nuclei of A549 cells (an immortalized human type II alveolar epithelial cell line). Exposure to TNF-alpha caused p65 and p50 to translocate into nuclei in a dose- and time-dependent manner. Preincubation with dexamethasone for 24 h or 30 min, or with theophylline for 30 min before TNF-alpha stimulation attenuated the nuclear translocation of Rel A and p50. These findings suggest that theophylline, as well as dexamethasone, may inhibit translocation of NF-kappaB.
Tomita
K
K
Department of Thoracic Medicine, Imperial College School of Medicine, London, UK.
Chikumi
H
H
Tokuyasu
H
H
Yajima
H
H
Hitsuda
Y
Y
Matsumoto
Y
Y
Sasaki
T
T
D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D003907:Dexamethasone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005455:Fluorescent Antibody Technique; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D016328:NF-kappa B / Q000378:metabolism*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D013806:Theophylline / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D018931:Antineoplastic Agents, Hormonal; D016328:NF-kappa B; D010726:Phosphodiesterase Inhibitors; D014409:Tumor Necrosis Factor-alpha; D003907:Dexamethasone; D013806:Theophylline