Am. J. Respir. Cell Mol. Biol.,
Volume 18, Number 2, February, 1998 243-254
Identification of Glucocorticoid- and Adenovirus E1A-regulated Genes in
Lung Epithelial Cells by Differential Display
Tokuji
Matsuba,
Naoto
Keicho,
Yuji
Higashimoto,
Shaun
Granleese,
James C.
Hogg,
Shizu
Hayashi,
and
Gregory P.
Bondy
University of British Columbia Pulmonary Research Laboratory, St. Paul's Hospital, Vancouver, British Columbia, Canada
Adenovirus infection has been implicated in the pathogenesis of lung inflammatory diseases for which
glucocorticoids provide effective antiinflammatory treatment. In this study, the differential display assay
was used to identify messenger RNAs (mRNAs) differentially expressed in dexamethasone (1 µM for 24 h)-
treated A549 lung epithelial cells compared to A549 cells transfected with the adenoviral E1A gene.
Thirty-seven complimentary DNAs (cDNAs) (15 glucocorticoid-regulated, 22 adenovirus E1A-regulated)
were isolated. DNA sequence analysis showed that 35 of these were unique, 2 were identical with each other, and 3 were common to the glucocorticoid- and E1A-regulated groups. Genes identified included
those involved in transcription/translation, cytoskeletal/contractile element genes, metabolic enzyme genes, and genes associated with cell regulation/signal transduction. After further analysis of the isolated clones
by Northern blotting, ribonuclease protection, and semiquantitative RT-PCR (reverse transcriptase-polymerase chain reaction), 10 of the 14 glucocorticoid-regulated and one of the three common to both the adenovirus E1A- and glucocorticoid-regulated cDNAs were confirmed for this control of their expression.
We conclude that the strategy of identifying cDNAs regulated by both adenovirus E1A and glucocorticoids provides a promising approach for identifying genes that may be important in the pathogenesis of
lung inflammation and therefore targets for glucocorticoid treatment.
