Regulation of MUC5AC expression by NAD(P)H:quinone oxidoreductase 1.
Neutrophil elastase (NE), a potent neutrophil inflammatory mediator, increases MUC5AC mucin gene expression through undefined pathways involving reactive oxygen species. To determine the source of NE-generated reactive oxygen species, we used pharmacologic inhibitors of oxidoreductases to test whether they blocked NE-regulated MUC5AC mRNA expression. We found that dicumarol, an inhibitor of the NADP(H):quinone oxidoreductase 1 (NQO1), inhibited MUC5AC mRNA expression in A549 lung adenocarcinoma cells and primary normal human bronchial epithelial cells. We further tested the role of NQO1 in mediating NE-induced MUC5AC expression by inhibiting NQO1 expression using short interfering RNA (siRNA). Transfection with siRNA specific for NQO1 suppressed NQO1 expression and significantly abrogated MUC5AC mRNA expression. NE treatment caused lipid peroxidation in A549 cells; this effect was inhibited by pretreatment with dicumarol, suggesting that NQO1 also regulates oxidant stress in A549 cells after NE exposure. NE exposure increased NQO1 protein and activity levels; NQO1 expression and activity were limited to the cytosol and did not translocate to the plasma membrane. Our results indicate that NQO1 has an important role as a key mediator of NE-regulated oxidant stress and MUC5AC mucin gene expression.
Zheng
Shuo
S
Department of Pediatrics, Duke University Medical Center, Durham, NC 27710, USA.
Byrd
Angela S
AS
Fischer
Bernard M
BM
Grover
Amy R
AR
Ghio
Andrew J
AJ
Voynow
Judith A
JA
D000230:Adenocarcinoma; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D006801:Humans; D019272:Leukocyte Elastase / Q000378:metabolism*; D008175:Lung Neoplasms; D055271:Mucin 5AC; D009077:Mucins / Q000235:genetics*; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000276:immunology / Q000378:metabolism*; D011808:Quinone Reductases / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic
C524678:MUC5AC protein, human; D055271:Mucin 5AC; D009077:Mucins; D012333:RNA, Messenger; C041831:NADH dehydrogenase (quinone); D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; D011808:Quinone Reductases; D019272:Leukocyte Elastase