Poly-L-lysine (PLL) was reported to suppress mucin release from airway goblet cells during 30 min treatment period. In this study, we investigated whether PLL consistently suppresses mucin release from cultured airway goblet cells during 24 h after 30 min treatment and also specifically suppresses the release of mucin without any effects on the other releasable glycoproteins. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24 h and chased for 30 min in the presence of varying concentrations of PLL to assess the effects on 3H-mucin release and on the total elution profile of the treated culture medium. The total mucin content during 24 h after 30 min treatment of PLL was assesed to investigate the consistency of effects. PLL did not affect the release of the other releasable glycoproteins whose molecular weights were less than mucin, and decreased the total mucin content during 24 h after 30 min treatment. We conclude that PLL can specifically suppress mucin release from cultured airway goblet cells and the suppression on mucin release is consistent. This finding suggests that PLL might be used as a specific airway mucin-regulating agent by directly acting on airway mucin-secreting cells.