The effect of insulin on glucose transport in skeletal muscle, cardiac muscle and adipose tissue has long been known (1), but effects on the human bronchial epithelial cell line (H441) have not been investigated. The presence of the facilitative glucose transporter (GLUT2) in H441 cells has been identified, but the insulin-sensitive GLUT4 transporter has not been fully confirmed (2). The aim of this study was to investigate the effect of insulin on glucose uptake in H441 cells in order to help identify the presence of insulin-sensitive glucose transporters in these cells. H441 monolayers were cultured on porous filters at air interface and uptake studies were performed using 10 mM glucose with radiolabelled [³H]-D-Glucose. Insulin (0-7 μM) was added either to the basolateral side or to the apical side of the monolayer 1 hour prior to glucose uptake being measured. Inhibition of glucose transport with 1 mM phloretin (GLUT inhibitor) was also studied. Results are expressed as mean ± standard error of the mean; treatments were compared using unpaired student’s t-test. Basolateral glucose uptake was 52.14 ± 19.84 nmol/mg protein with no insulin present. Maximal uptake was 484.46 ± 131.97 nmol/mg protein when 1.7 μM insulin was added to the basolateral side of the monolayer (p<0.05, n=5-9). The basolateral insulin concentration dose response effect was fitted with a sigmoidal curve with a Hill slope of 66.84 and a calculated EC₅₀ of 1.37 μM. Glucose uptake from the apical side of the monolayer was found to be less than basolateral uptake. With no insulin, glucose uptake was 18.05 ± 5.47 nmol/mg protein and maximal uptake was 60.04 ± 10.47 nmol/mg protein when 3.4 μM insulin was added to the apical chamber (p<0.05, n=4-8). The apical concentration response to insulin was also sigmoidal but was different to basolateral uptake with a Hill slope of 5.4 and a calculated EC₅₀ of 2.09 μM. In the presence of 1.7 μM insulin, inhibition with phloretin significantly reduced uptake from 432.68 ± 31.73 to 53.74 ± 9.55 nmol/mg protein (p<0.001, n=3). In conclusion, insulin stimulated basolateral and apical glucose uptake across H441 cell monolayers. These data indicate that insulin-sensitive glucose transporter(s) are present in the membranes of airway epithelial cells and inhibition with phloretin indicates that a significant component of insulin-sensitive uptake is via GLUT transporters.