Increase of bradykinin-stimulated arachidonic acid release in a delta F508 cystic fibrosis epithelial cell line.
Modification of chloride conductance by bradykinin in epithelial cells has been attributed to an activation of protein kinase A resulting from adenylcyclase stimulation by arachidonic acid cyclooxygenase products. The results presented here compare tracheal epithelial cell lines from one control and two cystic fibrosis patients which were immortalized by transfection with the SV40 large T oncogene. The three cell lines presented the same arachidonic acid content, turnover and mobilisation under basal conditions. Bradykinin stimulated the release of arachidonic acid and the synthesis of cyclooxygenase derivatives (mainly PGE2). The cell line from the cystic fibrosis patient bearing a phenylalanine 508 deletion, which is the major form of the disease, showed a higher bradykinin-induced arachidonic acid release than either control cells or cells from a patient presenting a minor form of the disease. This higher sensitivity suggests a dysregulation of phospholipase A2 stimulation in cystic fibrosis cells and was confirmed on non-immortalized tracheal epithelial cells in primary culture and on skin fibroblasts from patients bearing the same mutation. This defect is associated with a potentiation of cholera toxin pretreatment on cAMP content of delta F508 cell line. The impaired control of arachidonic acid release cannot be attributed to an increased number of bradykinin binding sites, since this increase was similar in the two cystic fibrosis cell lines.
Levistre
R
R
URA CNRS 1283, Faculté de Médecine Saint Antoine, Université Pierre et Marie Curie, Paris, France.
Lemnaouar
M
M
Rybkine
T
T
Béréziat
G
G
Masliah
J
J
D016718:Arachidonic Acid / Q000378:metabolism*; D001920:Bradykinin / Q000494:pharmacology*; D002460:Cell Line / Q000187:drug effects / Q000378:metabolism; D002772:Cholera Toxin; D003470:Culture Media / Q000032:analysis; D000242:Cyclic AMP / Q000032:analysis; D003550:Cystic Fibrosis / Q000235:genetics / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D007473:Ion Channels / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D009154:Mutation; D010743:Phospholipids / Q000378:metabolism; D011796:Quinacrine / Q000494:pharmacology; D012867:Skin / Q000378:metabolism; D014132:Trachea / Q000378:metabolism; D010566:Virulence Factors, Bordetella
D003470:Culture Media; D007473:Ion Channels; D008565:Membrane Proteins; D010743:Phospholipids; D010566:Virulence Factors, Bordetella; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D016718:Arachidonic Acid; D002772:Cholera Toxin; D000242:Cyclic AMP; D011796:Quinacrine; D001920:Bradykinin