Antisense oligodeoxynucleotide to the cystic fibrosis transmembrane conductance regulator inhibits cyclic AMP-activated but not calcium-activated cell volume reduction in a human pancreatic duct cell line.
Cystic fibrosis (CF) is characterized by a defect in cAMP-regulated chloride channels in epithelial cells. The CF gene product CF transmembrane conductance regulator (CFTR) is expressed in the apical membrane of pancreatic duct cells, and mutant CFTR accounts for the pathology in the CF pancreas. PANC 1, a pancreatic duct cell line, has not been considered a good model for studying CFTR and pancreatic chloride transport because CFTR mRNA and protein are undetectable using standard methods. Using electronic cell sizing and cell volume reduction under isotonic conditions, PANC 1 cells were found to possess both cAMP and calcium-activated chloride conductances. Using CFTR antisense oligodeoxynucleotides, the cAMP-activated conductance could be specifically inhibited in a concentration- and time-dependent manner. These findings demonstrate that PANC 1 cells express CFTR and a CFTR-independent calcium-activated chloride channel. With electronic cell sizing and CFTR antisense oligodeoxynucleotides, PANC 1 cells can provide an ideal system for the study of pancreatic duct cell physiology and pathophysiology with respect to the role of CFTR in the pancreas. These findings also suggest that antisense oligodeoxynucleotides may provide a more sensitive yet highly specific means of detecting low levels of expression of CFTR than currently available.
Kopelman
H
H
Division of Gastroenterology, Montreal Children's Hospital-McGill University Research Institute, Quebec, Canada.
Gauthier
C
C
Bornstein
M
M
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology*; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D002712:Chlorides / Q000378:metabolism*; D005576:Colforsin / Q000494:pharmacology*; D000242:Cyclic AMP / Q000378:metabolism* / Q000494:pharmacology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004305:Dose-Response Relationship, Drug; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D007700:Kinetics; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D010179:Pancreas / Q000187:drug effects / Q000502:physiology*; D013997:Time Factors
C505032:CFTR protein, human; D002712:Chlorides; D008565:Membrane Proteins; D016376:Oligonucleotides, Antisense; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005576:Colforsin; D015124:8-Bromo Cyclic Adenosine Monophosphate; D015759:Ionomycin; D000242:Cyclic AMP; D002118:Calcium