Inhibitory mechanism on NF-kB transactivation by dexamethasone in pulmonary epithelial cells / 결핵
Tuberculosis and Respiratory Diseases
;
: 682-698, 2000.
Artigo
em Coreano
| WPRIM
| ID: wpr-44265
ABSTRACT
Glucocorticoid receptor (GR) functions as a suppressor of inflammation by inhibiting the expression of many cytokine gene activated by NF-κB. The goal of this study is to investigate the mechanism by which GR repress NF-κB activation in lung epithelial cells. We used A549 and BEAS-2B lung epithelial cell lines. Using IgGκ-NF-κB luciferase reporter gene construct, we found that dexamethasone significantly suppressed TNF-α-induced NF-κB activation and the overexpression of GR showed dose-dependent reduction of TNF-α-induced NF-κB activity in both cell lines. However, DNA binding of NF-κB induced by TNF-α in electromobility shift assay was not inhibited by dexamethasone. Super shift assay with anti-p65 antibody demonstrated the existence of p65 in NF-κB complex induced by TNF-α Western blot showed that IκBα degradation induced by TNF-α was not affected by dexamethasone and IκBκ was not induced by dexamethasone, neither. To evaluate p65 specific transactivation, we adopted co-transfection study of Ga14-p65TA1 or TA2 fusion protein expression system together with 5xGa14-luciferase vector. Co-transfection of GR with Ga14-p65TA1 or TA2 repressed luciferase activity profoundly to the level of 10-20% of p65TA1- or TA2-induced transcriptional activity. And this transrepressional effect was abolished by co-transfection of CBP or SRC-1 expression vectors. These results suggest that Gr-mediated transrepression of NF-κB in lung epithelial cells is through competing for binding to limiting amount of transcriptional coactivators, CBP or SRC-1.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
DNA
/
Dexametasona
/
Receptores de Glucocorticoides
/
Ativação Transcricional
/
Linhagem Celular
/
Western Blotting
/
NF-kappa B
/
Genes Reporter
/
Células Epiteliais
/
Inflamação
Idioma:
Coreano
Revista:
Tuberculosis and Respiratory Diseases
Ano de publicação:
2000
Tipo de documento:
Artigo
Similares
MEDLINE
...
LILACS
LIS