title: Cigarette smoke alters non-neuronal cholinergic system components inducing MUC5AC production in the H292 cell line.
abstract: Cigarette smoke extract (CSE) affects the expression of Choline Acetyl-Transferase (ChAT), muscarinic acetylcholine receptors, and mucin production in bronchial epithelial cells. Mucin 5AC (MUC5AC), muscarinic acetylcholine receptor M3, ChAT expression, acetylcholine levels and acetylcholine binding were measured in a human pulmonary mucoepidermoid carcinoma cell line (H292) stimulated with CSE. We performed ChAT/RNA interference experiments in H292 cells stimulated with CSE to study the role of ChAT/acetylcholine in MUC5AC production. The effects of Hemicholinium-3 (HCh-3) (50 μM) (a potent and selective choline uptake blocker) and Tiotropium bromide (Spiriva(®)) (100 nM), alone or in combination with Salmeterol (SL) and Fluticasone propionate (FP), were tested in this model. MUC5AC, muscarinic acetylcholine receptor M3, ChAT, acetylcholine expression and acetylcholine binding significantly increased in H292 cells stimulated with CSE (5%) compared to untreated cells. HCh-3 reduced acetylcholine binding and MUC5AC production in H292 cells stimulated with CSE. ChAT/RNA interference eliminated the effect of CSE on MUC5AC production. FP reduced ChAT and acetylcholine binding in unstimulated cells, while showing a partial effect in CSE stimulated cells. SL increased the ChAT expression and acetylcholine binding in H292 cells stimulated with or without CSE. Tiotropium, alone or together with FP and SL, reduced acetylcholine binding and MUC5AC production in H292 cells stimulated with CSE. CSE affects the ChAT/acetylcholine expression, increasing MUC5AC production in H292 cells. Pharmacological treatment with anticholinergic drugs reduces the secretion of MUC5AC generated by autocrine acetylcholine activity in airway epithelial cells.
authors.affiliation: Institute of Biomedicine and Molecular Immunology "A. Monroy" (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Angela Marina
authors.initials: AM
authors.lastname: Montalbano
authors.affiliation: Institute of Biomedicine and Molecular Immunology "A. Monroy" (IBIM), Italian National Research Council (CNR), Palermo, Italy; Dipartimento Biomedico di Medicina Interna e Specialistica (Di.Bi.M.I.S.), Sezione di Pneumologia, University of Palermo, Palermo, Italy.
authors.forename: Giusy Daniela
authors.initials: GD
authors.lastname: Albano
authors.affiliation: Institute of Biomedicine and Molecular Immunology "A. Monroy" (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Anzalone
authors.affiliation: Institute of Biomedicine and Molecular Immunology "A. Monroy" (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Bonanno
authors.affiliation: Institute of Biomedicine and Molecular Immunology "A. Monroy" (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Loredana
authors.initials: L
authors.lastname: Riccobono
authors.affiliation: Institute of Biomedicine and Molecular Immunology "A. Monroy" (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Di Sano
authors.affiliation: Institute of Biomedicine and Molecular Immunology "A. Monroy" (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Rosalia
authors.initials: R
authors.lastname: Gagliardo
authors.affiliation: Institute of Biomedicine and Molecular Immunology "A. Monroy" (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Liboria
authors.initials: L
authors.lastname: Siena
authors.affiliation: Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany.
authors.forename: Michael Paul
authors.initials: MP
authors.lastname: Pieper
authors.affiliation: Institute of Biomedicine and Molecular Immunology "A. Monroy" (IBIM), Italian National Research Council (CNR), Palermo, Italy.
authors.forename: Mark
authors.initials: M
authors.lastname: Gjomarkaj
authors.affiliation: Institute of Biomedicine and Molecular Immunology "A. Monroy" (IBIM), Italian National Research Council (CNR), Palermo, Italy. Electronic address: profita@ibim.cnr.it.
authors.forename: Mirella
authors.initials: M
authors.lastname: Profita
mesh_terms: D000109:Acetylcholine / Q000378:metabolism*; D000420:Albuterol / Q000031:analogs & derivatives / Q000494:pharmacology; D000730:Androstadienes / Q000494:pharmacology; D001980:Bronchi / Q000166:cytology; D001993:Bronchodilator Agents / Q000494:pharmacology; D045744:Cell Line, Tumor; D002795:Choline O-Acetyltransferase / Q000235:genetics / Q000378:metabolism*; D018680:Cholinergic Antagonists / Q000494:pharmacology; D045424:Complex Mixtures / Q000494:pharmacology*; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D000068298:Fluticasone; D006426:Hemicholinium 3 / Q000494:pharmacology; D006801:Humans; D055271:Mucin 5AC / Q000378:metabolism*; D014179:Neurotransmitter Uptake Inhibitors / Q000494:pharmacology; D034622:RNA Interference; D043587:Receptor, Muscarinic M3 / Q000378:metabolism; D000068299:Salmeterol Xinafoate; D012602:Scopolamine Derivatives / Q000494:pharmacology; D012906:Smoke; D000069447:Tiotropium Bromide; D014026:Nicotiana
chemical_list: D000730:Androstadienes; D001993:Bronchodilator Agents; D018680:Cholinergic Antagonists; D045424:Complex Mixtures; C524678:MUC5AC protein, human; D055271:Mucin 5AC; D014179:Neurotransmitter Uptake Inhibitors; D043587:Receptor, Muscarinic M3; D012602:Scopolamine Derivatives; D012906:Smoke; D006426:Hemicholinium 3; D000068299:Salmeterol Xinafoate; D000068298:Fluticasone; D002795:Choline O-Acetyltransferase; D000109:Acetylcholine; D000420:Albuterol; D000069447:Tiotropium Bromide
keywords: Anticholinergic drugs; Choline Acetyl-Transferase; Cigarette smoke; Mucin production.