title: Insulin stimulates production of glycoconjugate layers on the cell surface of gastric surface mucous cell line GSM06.
abstract: The mechanism of regulation of mucus production in the gastric mucosa remains unclear. Recently, we established a gastric surface mucous cell line GSM06, which produces periodic acid-Shiff (PAS)-positive glycoconjugate (mucus) layers on the cell surface, from transgenic mice harboring a temperature-sensitive simian virus 40 large T-antigen gene. In this study, GSM06 cells were examined for its production of PAS-positive glycoconjugate layers to acid secretagogues and growth factors. The cells were cultured at nonpermissive temperature (39 degrees C) for 3-18 days and stained with PAS. Insulin (1-30 microg/ml; 0.29-8.6 microM) time- and dose-dependently increased production of glycoconjugates on the cell surface. When glycoconjugate layers produced by stimulation of insulin (3-30 microg/ml; 0.86-8.6 microM) were removed from the cell surface of GSM06 cells by a mild trypsin treatment, PAS-positive materials were remarkably decreased (day 18). In addition, morphological findings indicate that a high concentration of insulin (30 microg/ml; 8.6 microM) produced thick PAS-positive glycoconjugate layers just like normal gastric surface mucosa on the cell surface on day 18. In contrast, histamine (0.1-100 microM), carbachol (0.1-100 microM), gastrin-17 (0.1-100 nM), epidermal growth factor (0.01-10 ng/ ml; 1.7-1,700 pM), transforming growth factor-alpha (0.01-10 ng/ml; 1.8-1,800 pM), and fetal bovine serum (1-10%) did not increase glycoconjugate production. These findings suggest that insulin is a stimulator of glycoconjugate production, and stimulates production of glycoconjugate layers on the cell surface in the gastric surface mucous cell line GSM06.
authors.affiliation: Molecular Biology Research Laboratory, Daiichi Pharmaceutical Co., Ltd., Tokyo, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Tabuchi
authors.forename: N
authors.initials: N
authors.lastname: Sugiyama
authors.forename: T
authors.initials: T
authors.lastname: Horiuchi
authors.forename: K
authors.initials: K
authors.lastname: Furuhama
authors.forename: M
authors.initials: M
authors.lastname: Furusawa
mesh_terms: D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005753:Gastric Mucosa / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005755:Gastrins / Q000494:pharmacology; D006001:Glycoconjugates / Q000096:biosynthesis*; D006632:Histamine / Q000494:pharmacology; D007004:Hypoglycemic Agents / Q000008:administration & dosage / Q000494:pharmacology*; D007328:Insulin / Q000008:administration & dosage / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018722:Nicotinic Agonists / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013268:Stimulation, Chemical; D016211:Transforming Growth Factor alpha / Q000494:pharmacology
chemical_list: D005755:Gastrins; D006001:Glycoconjugates; D007004:Hypoglycemic Agents; D007328:Insulin; D018722:Nicotinic Agonists; D016211:Transforming Growth Factor alpha; D012710:Serum Albumin, Bovine; C013550:gastrin 17; D004815:Epidermal Growth Factor; D006632:Histamine; D002217:Carbachol