title: [Cellular expression of CFTR in cystic fibrosis: defective cyclic AMP-dependent regulation of glycoconjugate secretion in cystic fibrosis fetal tracheal epithelial cells transfected by SV40 large T oncogene].
abstract: Epithelial tracheal cells isolated from two fetuses with cystic fibrosis (CF) and non-CF fetus (control) were transfected with a plasmid vector recombined with the large T oncogene of SV40. All transfected cells expressed SV40 antigen and exhibited an epithelial morphology (junctional complex, cytokeratins). CFT cells retained the mutations of the CF gene, one heterozygous for the S549N/N1303K substitutions (CFT-1 cells), the other homozygous for the deletion delta F508 (CFT-2 cells). Accordingly, these CFT cells exhibited the defective beta-adrenergic regulation of chloride conductance. We compared the responsiveness of control (NT-1 cells) and CF cells (CFT-1 and CFT-2 cells) to agonists of the protein kinase A (PKA)-dependent pathway for stimulation of glycoconjugate secretion. We show that the isoproterenol (10(-5) M) and forskolin (10(-5) M) markedly increased the cAMP content and the PKA activity of all three cell lines. In contrast, these effectors produced an increase in glycoconjugate secretion in control cells, but not in CFT-1 and CFT-2 cells. In conclusion, our results indicate that CFT cells do not respond to agonists of the PKA-dependent pathway for stimulation of both glycoconjugate secretion and chloride transport, which suggests the involvement of CFTR in these two processes.
authors.affiliation: Inserm U181, Laboratoire de Biochimie-Biologie cellulaire, Faculté de Médecine Saint-Antoine, Paris.
authors.forename: J
authors.initials: J
authors.lastname: Picard
authors.forename: M
authors.initials: M
authors.lastname: Lemnaouar
authors.forename: A
authors.initials: A
authors.lastname: Paul
mesh_terms: D002478:Cells, Cultured; D000242:Cyclic AMP / Q000502:physiology*; D003550:Cystic Fibrosis / Q000235:genetics / Q000473:pathology / Q000503:physiopathology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004848:Epithelium / Q000378:metabolism / Q000473:pathology; D005333:Fetus / Q000473:pathology; D005814:Genes, Viral; D006001:Glycoconjugates / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000096:biosynthesis*; D009857:Oncogenes; D013539:Simian virus 40 / Q000235:genetics; D014132:Trachea / Q000378:metabolism* / Q000473:pathology; D014162:Transfection
chemical_list: C505032:CFTR protein, human; D006001:Glycoconjugates; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000242:Cyclic AMP