Goblet cell hyperplasia is a common feature of chronic obstructive pulmonary disease (COPD) airways, but the mechanisms that underlie this epithelial remodelling in COPD are not understood. Based on our previous finding of hypoxia‐inducible factor‐1α (HIF‐1α) nuclear localization in large airways from patients with COPD, we investigated whether hypoxia‐inducible signalling could influence the development of goblet cell hyperplasia. We evaluated large airway samples obtained from 18 lifelong non‐smokers and 13 former smokers without COPD, and 45 former smokers with COPD. In these specimens, HIF‐1α nuclear staining occurred almost exclusively in COPD patients in areas of airway remodelling. In COPD patients, 93.2 ± 3.9% (range 65–100%) of goblet cells were HIF‐1α positive in areas of goblet cell hyperplasia, whereas nuclear HIF‐1α was not detected in individuals without COPD or in normal‐appearing pseudostratified epithelium from COPD patients. To determine the direct effects of hypoxia‐inducible signalling on epithelial cell differentiation in vitro, human bronchial epithelial cells (HBECs) were grown in air–liquid interface cultures under hypoxia (1% O₂) or following treatment with a selective HIF‐1α stabilizer, (2R)‐[(4‐biphenylylsulphonyl)amino]‐N‐hydroxy‐3‐phenyl‐propionamide (BiPS). HBECs grown in hypoxia or with BiPS treatment were characterized by HIF‐1α activation, carbonic anhydrase IX expression, mucus‐producing cell hyperplasia and increased expression of MUC5AC. Analysis of signal transduction pathways in cells with HIF‐1α activation showed increased ERK1/2 phosphorylation without activation of epidermal growth factor receptor, Ras, PI3K‐Akt or STAT6. These data indicate an important effect of hypoxia‐inducible signalling on airway epithelial cell differentiation and identify a new potential target to limit mucus production in COPD. Copyright © 2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.