INVITRO EFFECTS OF DRUGS ON PRODUCTION OF MUCINS IN RABBIT TRACHEAL EPITHELIAL-CELLS EXPRESSING MUCIN GENE - A MODEL SYSTEM FOR STUDYING UPPER AIRWAY RESPIRATORY-DISEASES

Rabbit tracheal epithelial cells, cultured on collagen-coated dishes in serum-free and hormone-supplemented medium, were found to incorporate [H-3]glucosamine into high-molecular-weight components that were secreted in the medium. The chemical analysis of the secreted products resulted in a profile that resembled that of mucous glycoproteins (mucins). When examined by dot blot analysis, the total RNA isolated from these cells hybridized to an antisense 30-mer oligonucleotide corresponding to a rat intestine mucin peptide sequence, indicating that mucin gene was expressed in these cell lines. Lung and liver tissues of rabbit did not express this gene. Transmission electron microscopy exhibited secretory granules in these cells. The incorporation of [H-3]glucosamine into mucins was inhibited by three aryl-N-acetyl-galactosaminides and a chemical carcinogen, N-nitroso-N-ethyl urea, whereas 5-azacytidine enhanced the proliferation of cells as well as the radiolabeling of mucins. Parasympathetic agent (pilocarpine), cholinergic antagonist (atropine), and beta-adrenergic agonist (isoproterenol) alone have little effect on the secretion of mucins. The cholinergic agonist, methacholine, was found to increase the production of mucins and addition of atropine to the medium before methacholine blocked this stimulation. Histamine was found to stimulate mucin production in these cells.