1718974

Source:http://linkedlifedata.com/resource/pubmed/id/1718974

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Predicate	Object
rdf:type	pubmed:Citation
lifeskim:mentions	umls-concept:C0007634, umls-concept:C0010674, umls-concept:C0029016, umls-concept:C0038975, umls-concept:C0205245, umls-concept:C0226890, umls-concept:C0441587, umls-concept:C1880022
pubmed:issue	31
pubmed:dateCreated	1991-12-13
pubmed:abstractText	Intestinal epithelial cells were isolated from a fetus with cystic fibrosis (CF) and transfected with a plasmid vector recombined with the ori- mutant of SV40. A population of proliferative cells was then subcloned and designated as CFI-3. These cells had a doubling time of 24 h and were maintained in culture for up to 25 passages. At passage 8, CFI-3 cells did not produce any tumors in nude mice. Northern blot and immunofluorescence studies indicated that the extended lifespan of CFI-3 cells results in genomic insertion of SV40 LT. Intestinal CFI-3 cells are epithelial, according to the expression of the human cytokeratin 18 gene and poorly differentiated by phase-contrast and electron microscopy. Functional membrane receptors activated by vasoactive intestinal peptide (VIP), its natural analogue pituitary adenylate cyclase activating peptide (PACAP-38), and isoproterenol were observed in CFI-3 cells. Restriction fragment length polymorphism analysis of the PstI KM19 site revealed that the cftr locus was identical in the chorionic villi and in CFI-3 cells. The manifestation of CF in this family was not related to the common mutation delta F508, since this fetus was heterozygous for the substitutions S549N and N1303K. Chloride transport, assessed by the 125I efflux, was induced in CFI-3 cells by the calcium inophore ionomycin, but not by the adenylate cyclase activator forskolin, and was inhibited by the chloride channel blocker 5-nitro-2-(3-phenylpropylamino)benzoic acid. These results were confirmed in patch clamp studies in which the cpt cAMP analogue failed to stimulate membrane currents, while the calcium ionophore ionomycin stimulated inward currents. We conclude that intestinal CFI-3 cells retain the CF phenotype relating to defective regulation of Cl- channels, and therefore constitute a suitable model, 1) for elucidating the function of CFTR protein, 2) developing new therapeutic agents, and 3) correcting the CF defect by gene replacement therapy in vitro.
pubmed:language	eng
pubmed:journal	http://linkedlifedata.com/resource/pubmed/journal/2985121R
pubmed:citationSubset	IM
pubmed:chemical	http://linkedlifedata.com/resource/pubmed/chemical/ADCYAP1 protein, human, http://linkedlifedata.com/resource/pubmed/chemical/Antigens, Polyomavirus Transforming, http://linkedlifedata.com/resource/pubmed/chemical/CFTR protein, human, http://linkedlifedata.com/resource/pubmed/chemical/Chloride Channels, http://linkedlifedata.com/resource/pubmed/chemical/Cyclic AMP, http://linkedlifedata.com/resource/pubmed/chemical/Cystic Fibrosis Transmembrane..., http://linkedlifedata.com/resource/pubmed/chemical/Forskolin, http://linkedlifedata.com/resource/pubmed/chemical/Ionomycin, http://linkedlifedata.com/resource/pubmed/chemical/Isoproterenol, http://linkedlifedata.com/resource/pubmed/chemical/Membrane Proteins, http://linkedlifedata.com/resource/pubmed/chemical/Neuropeptides, http://linkedlifedata.com/resource/pubmed/chemical/Pituitary Adenylate..., http://linkedlifedata.com/resource/pubmed/chemical/Potassium Channels, http://linkedlifedata.com/resource/pubmed/chemical/Vasoactive Intestinal Peptide
pubmed:status	MEDLINE
pubmed:month	Nov
pubmed:issn	0021-9258
pubmed:author	pubmed-author:BarbryPP, pubmed-author:ChampignyGG, pubmed-author:ChastreEE, pubmed-author:Di GioiaYY, pubmed-author:EmamiSS, pubmed-author:FanenPP, pubmed-author:GespachCC, pubmed-author:MornetEE, pubmed-author:Simon-BouyBB
pubmed:issnType	Print
pubmed:day	5
pubmed:volume	266
pubmed:owner	NLM
pubmed:authorsComplete	Y
pubmed:pagination	21239-46
pubmed:dateRevised	2006-11-15
pubmed:meshHeading	pubmed-meshheading:1718974-Antigens, Polyomavirus Transforming, pubmed-meshheading:1718974-Base Sequence, pubmed-meshheading:1718974-Cell Line, pubmed-meshheading:1718974-Cell Transformation, Viral, pubmed-meshheading:1718974-Chloride Channels, pubmed-meshheading:1718974-Cyclic AMP, pubmed-meshheading:1718974-Cystic Fibrosis, pubmed-meshheading:1718974-Cystic Fibrosis Transmembrane Conductance Regulator, pubmed-meshheading:1718974-Forskolin, pubmed-meshheading:1718974-Gene Expression, pubmed-meshheading:1718974-Humans, pubmed-meshheading:1718974-Intestinal Mucosa, pubmed-meshheading:1718974-Ionomycin, pubmed-meshheading:1718974-Isoproterenol, pubmed-meshheading:1718974-Membrane Proteins, pubmed-meshheading:1718974-Molecular Sequence Data, pubmed-meshheading:1718974-Neuropeptides, pubmed-meshheading:1718974-Pituitary Adenylate Cyclase-Activating Polypeptide, pubmed-meshheading:1718974-Potassium Channels, pubmed-meshheading:1718974-Transfection, pubmed-meshheading:1718974-Vasoactive Intestinal Peptide
pubmed:year	1991
pubmed:articleTitle	Functional insertion of the SV40 large T oncogene in cystic fibrosis intestinal epithelium. Characterization of CFI-3 cells.
pubmed:affiliation	Institut National de la Santé et de la Recherche Médicale U.55, Unité de Recherches sur les Peptides Neurodigestifs et le Diabète, Paris, France.
pubmed:publicationType	Journal Article, In Vitro, Research Support, Non-U.S. Gov't