Identification of novel inducible genes in airway epithelium.
DNA differential display analysis (DD-PCR) was utilized to identify genes that are expressed in airway epithelium and are relevant to airway inflammation; cytokine-mediated induction of gene expression and inhibition of that induction by glucocorticoids were the criteria for selection. The IB3-1 cell line was cultured in the presence of tumor necrosis factor-alpha (TNF-alpha), dexamethasone, or dimethyl sulfoxide (DMSO) as a control, and analyzed via DD-PCR and Northern blot analyses. With this approach, two TNF-alpha-inducible and dexamethasone (DEX)-sensitive expressed sequence tags (EST8 and EST19) were identified. In IB3-1 cells, TNF-alpha increased messenger RNA (mRNA) expression of EST8 (34%, P < or = 0.005) and EST19 (41%, P < or = 0.01), whereas dexamethasone reduced this expression to resting levels. This pattern of mRNA expression was also observed in normal human bronchial epithelial cells (EST8: 21%, P < or = 0.009; EST19: 11%, P < or = 0.02) and in the basophil leukemia cell line KU812 (EST8: 34%, P < or = 0.01). Through basic local alignment search tool (BLAST) analysis, it was determined that these ESTs exhibited significant homology with the monomeric G protein rhoC (EST8: 100% homology, P = 1.6 x 10(-100)) and the UFO tyrosine kinase receptor (EST19: 86% homology, 5.3 x 10(-28).
Schwiebert
L M
LM
Department of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.
Mooney
J L
JL
Van Horn
S
S
Gupta
A
A
Schleimer
R P
RP
D001483:Base Sequence; D001980:Bronchi / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D016207:Cytokines / Q000494:pharmacology; D004247:DNA; D003626:Data Display; D003907:Dexamethasone / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004848:Epithelium / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D005938:Glucocorticoids / Q000494:pharmacology; D006801:Humans; D007249:Inflammation; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000096:biosynthesis; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D016324:Sequence Tagged Sites; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D016207:Cytokines; D005938:Glucocorticoids; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D003907:Dexamethasone; D004247:DNA; D004121:Dimethyl Sulfoxide