Epidermal growth factor receptors (EGFRs) are increased in airway smooth muscle in asthma, which may contribute to both their hyperproliferation and hypercontractility. Lysophosphatidic acid (LPA) is a candidate pathologic agent in asthma and other airway diseases, and LPA up-regulates EGFRs in human airway smooth muscle (HASM) cells. We tested whether therapeutic glucocorticoids and/or beta-2 adrenergic receptor (₂AR) agonists also alter EGFR binding in HASM cells. Exposure to glucocorticoids for 24 hr induced a two-fold increase in EGFR binding similar to that with LPA; fluticasone was markedly more potent than dexamethasone. The increase in EGFR binding by glucocorticoids required 24-hr exposure, consistent with transcription-mediated effects. Though the increase in EGFR binding was blocked by the protein synthesis inhibitor cycloheximide for LPA, fluticasone, and dexamethasone, only LPA induced a significant increase in EGFR protein expression detected by immunoblotting. In contrast to the increased binding induced by the glucocorticoids, the ₂AR agonists isoproterenol, albuterol and salmeterol all induced a decrease in EGFR binding. ₂AR agonist effects were multiphasic, with an initial decline at 2-4 hr that reversed by 6 hr and a second somewhat greater decrease by 18-24 hr. In cells pretreated with glucocorticoids, the decreases in EGFR binding by subsequent ₂AR treatment were not statistically significant; glucocorticoid up-regulation of EGFRs also prevented further increases by LPA. Similar increases by glucocorticoids and decreases by ₂AR agonists were found in HFL-1 lung fibroblasts. These complex and opposing effects of clinically relevant glucocorticoids and ₂AR agonists on airway mesenchymal cell EGFRs likely contribute to their overall therapeutic profile in the diseased airway.