Abstract
Continuous epithelial cell lines from individuals with cystic fibrosis (CF) and normal controls are required to understand the genetic and cellular defects in CF. We used retroviruses to transduce SV40 large T antigen into nasal epithelial cells. Transformed continuous cell lines were isolated that expressed epithelial markers, cytokeratin, and tight junctions. Northern blot analysis shows that all of the cell lines express the putative CF gene mRNA. Studies of transepithelial electrolyte transport show that CF and normal cell lines develop a transepithelial electrical resistance. Normal but not CF cell lines secreted Cl- in response to agonists that increase cellular levels of adenosine 3',5'-cyclic monophosphate (cAMP) (isoproterenol, forskolin, and a membrane-permeant analogue of cAMP) or in response to a tumor-promoting phorbol ester that activates protein kinase C. In contrast, the Ca2(+)-elevating agonist bradykinin and the Ca2+ ionophore A23187 stimulated secretion in both normal and CF cell lines. The continuous cell lines we have produced maintain their proper phenotypes and will serve as useful tools in understanding the pathophysiology of CF.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Antigens, Polyomavirus Transforming / genetics
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Base Sequence
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Biomarkers
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Blood Proteins / genetics
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Bradykinin / pharmacology
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Calcimycin / pharmacology
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Calgranulin A
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Cell Line
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Cell Transformation, Neoplastic
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Cells, Cultured
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Chloride Channels
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Chlorides / metabolism
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Colforsin / pharmacology
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Cystic Fibrosis / genetics*
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Cystic Fibrosis / pathology
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Cystic Fibrosis / physiopathology
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Epithelial Cells
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Epithelium / physiology
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Genotype
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Humans
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Ion Channels / drug effects
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Ion Channels / physiology
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Keratins / analysis
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Membrane Potentials / drug effects
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Membrane Proteins / physiology
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Molecular Sequence Data
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Nasal Mucosa / cytology
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Nasal Mucosa / pathology
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Nasal Mucosa / physiology
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Oligonucleotide Probes
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Phenotype
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Polymerase Chain Reaction
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Reference Values
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Retroviridae / genetics
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Simian virus 40 / genetics
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Tetradecanoylphorbol Acetate / pharmacology
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Theophylline / pharmacology
Substances
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Antigens, Polyomavirus Transforming
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Biomarkers
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Blood Proteins
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Calgranulin A
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Chloride Channels
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Chlorides
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Ion Channels
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Membrane Proteins
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Oligonucleotide Probes
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cystic fibrosis serum factor
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Colforsin
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Calcimycin
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Keratins
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Theophylline
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Tetradecanoylphorbol Acetate
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Bradykinin