E-GEOD-13168 - Transcription profiling of human airway smooth muscle cells after treatment with glucocorticoids and Protein Kinase A
Submitted on 13 October 2008, released on 13 November 2008, last updated on 10 June 2011
Glucocorticoids (GCs) and protein kinase A (PKA)-activating agents (beta-adrenergic receptor agonists) are mainstream asthma therapies based on their ability to prevent or reverse excessive airway smooth muscle (ASM) constriction. Their abilities to regulate another important feature of asthma - excessive ASM growth are poorly understood. Recent studies have suggested that GCs render agents of inflammation such as interleukin 1beta and tumor necrosis factor alpha mitogenic to ASM, via suppression of (antimitogenic) induced cyclooxygenase-2-dependent PKA activity. To further explore the mechanistic basis of these observations, we assessed the effects of epidermal growth factor and interleukin 1beta stimulation, and the modulatory effects of GC treatment and PKA inhibition, on the ASM transcriptome by microarray analysis. Experiment Overall Design: Human ASM cultures were generated from trachea of 4 unidentified donors and used in passage 5-8. These cells were used to generate cells stabely expressing a PKA inhibitor (PKI) or GFP (control), which were then grown to confluence, serum starved for 24h, and stimulated with IL-1b, EGF,or both (E+I) in presence or absence of Fluticasone (Flu, 30 min pretreatment). After 8h stimulation cells were washed and total RNA isolated using TRIzol as per manufacturer's protocol. Gene expression was analyzed in 4 different cultures derived from 4 different donors (for a total of 3-4 microarrays per condition).
transcription profiling by array, unknown experiment type