Liposome-mediated transfection of fetal lung epithelial cells: DNA degradation and enhanced superoxide toxicity.
Identifieur interne : 002720 ( Main/Exploration ); précédent : 002719; suivant : 002721Liposome-mediated transfection of fetal lung epithelial cells: DNA degradation and enhanced superoxide toxicity.
Auteurs : A K Tanswell [Canada] ; O. Staub ; R. Iles ; R. Belcastro ; J. Cabacungan ; L. Sedlackova ; B. Steer ; Y. Wen ; J. Hu ; H. O'BrodovichSource :
- The American journal of physiology [ 0002-9513 ] ; 1998.
Descripteurs français
- KwdFr :
- ADN (métabolisme), Animaux, Cellules cultivées, Cellules épithéliales (), Cellules épithéliales (cytologie), Cellules épithéliales (physiologie), Chloramphenicol O-acetyltransferase (biosynthèse), Chloramphenicol O-acetyltransferase (génétique), Composés d'ammonium quaternaire, Cytomegalovirus, Foetus, Gènes rapporteurs, Liposomes, Phosphatidyléthanolamine, Plasmides, Poumon (), Poumon (cytologie), Poumon (physiologie), Protéines recombinantes (biosynthèse), Rats, Superoxydes (toxicité), Survie cellulaire (), Transfection (), Vecteurs de médicaments, Vecteurs génétiques.
- MESH :
- biosynthèse : Chloramphenicol O-acetyltransferase, Protéines recombinantes.
- cytologie : Cellules épithéliales, Poumon.
- génétique : Chloramphenicol O-acetyltransferase.
- métabolisme : ADN.
- physiologie : Cellules épithéliales, Poumon.
- toxicité : Superoxydes.
- Animaux, Cellules cultivées, Cellules épithéliales, Composés d'ammonium quaternaire, Cytomegalovirus, Foetus, Gènes rapporteurs, Liposomes, Phosphatidyléthanolamine, Plasmides, Poumon, Rats, Survie cellulaire, Transfection, Vecteurs de médicaments, Vecteurs génétiques.
English descriptors
- KwdEn :
- Animals, Cell Survival (drug effects), Cells, Cultured, Chloramphenicol O-Acetyltransferase (biosynthesis), Chloramphenicol O-Acetyltransferase (genetics), Cytomegalovirus, DNA (metabolism), Drug Carriers, Epithelial Cells (cytology), Epithelial Cells (drug effects), Epithelial Cells (physiology), Fetus, Genes, Reporter, Genetic Vectors, Liposomes, Lung (cytology), Lung (drug effects), Lung (physiology), Phosphatidylethanolamines, Plasmids, Quaternary Ammonium Compounds, Rats, Recombinant Proteins (biosynthesis), Superoxides (toxicity), Transfection (methods).
- MESH :
- chemical , biosynthesis : Chloramphenicol O-Acetyltransferase, Recombinant Proteins.
- cytology : Epithelial Cells, Lung.
- drug effects : Cell Survival, Epithelial Cells, Lung.
- chemical , genetics : Chloramphenicol O-Acetyltransferase.
- chemical , metabolism : DNA.
- methods : Transfection.
- physiology : Epithelial Cells, Lung.
- chemical , toxicity : Superoxides.
- Animals, Cells, Cultured, Cytomegalovirus, Drug Carriers, Fetus, Genes, Reporter, Genetic Vectors, Liposomes, Phosphatidylethanolamines, Plasmids, Quaternary Ammonium Compounds, Rats.
Abstract
Cationic liposomes, 1:1 (mol/mol) 1,2-dioleoyldimethylammonium chloride-1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, were used to transfect primary cultures of distal rat fetal lung epithelial cells with pCMV4-based plasmids. A DNA-to-lipid ratio of 1:10 to 1:15 (wt/wt) optimized DNA uptake over a 24-h exposure. At a fixed DNA-to-lipid ratio of 1:15, chloramphenicol acetyltransferase (CAT) reporter gene expression declined at lipid concentrations > 2.5 nmol/cm2 cell surface area, whereas DNA uptake remained concentration dependent. CAT expression peaked 48 h after removal of the liposome-DNA complex, declining thereafter. Reporter gene expression was increased, and supercoiled cDNA degradation was reduced by the addition of 0.2 mM nicotinamide and 10 microM chloroquine. Rat fetal lung epithelial cells transfected with two different expression cassettes had an increased susceptibility to superoxide-mediated cytotoxicity. This could be attributed to a nonspecific delivery of exogenous DNA or some other copurified factor. The DNA-dependent increase in superoxide-mediated cytotoxicity, but not basal levels of cytotoxicity, was inhibited by the addition of 0.2 mM nicotinamide and 10 microM chloroquine.
DOI: 10.1152/ajplung.1998.275.3.L452
PubMed: 9728039
Affiliations:
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O'Brodovich</name></author></analytic><series><title level="j">The American journal of physiology</title><idno type="ISSN">0002-9513</idno><imprint><date when="1998" type="published">1998</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Cell Survival (drug effects)</term><term>Cells, Cultured</term><term>Chloramphenicol O-Acetyltransferase (biosynthesis)</term><term>Chloramphenicol O-Acetyltransferase (genetics)</term><term>Cytomegalovirus</term><term>DNA (metabolism)</term><term>Drug Carriers</term><term>Epithelial Cells (cytology)</term><term>Epithelial Cells (drug effects)</term><term>Epithelial Cells (physiology)</term><term>Fetus</term><term>Genes, Reporter</term><term>Genetic Vectors</term><term>Liposomes</term><term>Lung (cytology)</term><term>Lung (drug effects)</term><term>Lung (physiology)</term><term>Phosphatidylethanolamines</term><term>Plasmids</term><term>Quaternary Ammonium Compounds</term><term>Rats</term><term>Recombinant Proteins (biosynthesis)</term><term>Superoxides (toxicity)</term><term>Transfection (methods)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN (métabolisme)</term><term>Animaux</term><term>Cellules cultivées</term><term>Cellules épithéliales ()</term><term>Cellules épithéliales (cytologie)</term><term>Cellules épithéliales (physiologie)</term><term>Chloramphenicol O-acetyltransferase (biosynthèse)</term><term>Chloramphenicol O-acetyltransferase (génétique)</term><term>Composés d'ammonium quaternaire</term><term>Cytomegalovirus</term><term>Foetus</term><term>Gènes rapporteurs</term><term>Liposomes</term><term>Phosphatidyléthanolamine</term><term>Plasmides</term><term>Poumon ()</term><term>Poumon (cytologie)</term><term>Poumon (physiologie)</term><term>Protéines recombinantes (biosynthèse)</term><term>Rats</term><term>Superoxydes (toxicité)</term><term>Survie cellulaire ()</term><term>Transfection ()</term><term>Vecteurs de médicaments</term><term>Vecteurs génétiques</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Chloramphenicol O-Acetyltransferase</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Chloramphenicol O-acetyltransferase</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="cytologie" xml:lang="fr"><term>Cellules épithéliales</term><term>Poumon</term></keywords><keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Epithelial Cells</term><term>Lung</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Cell Survival</term><term>Epithelial Cells</term><term>Lung</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Chloramphenicol O-Acetyltransferase</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Chloramphenicol O-acetyltransferase</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>DNA</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Transfection</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ADN</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Cellules épithéliales</term><term>Poumon</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Epithelial Cells</term><term>Lung</term></keywords><keywords scheme="MESH" type="chemical" qualifier="toxicity" xml:lang="en"><term>Superoxides</term></keywords><keywords scheme="MESH" qualifier="toxicité" xml:lang="fr"><term>Superoxydes</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cells, Cultured</term><term>Cytomegalovirus</term><term>Drug Carriers</term><term>Fetus</term><term>Genes, Reporter</term><term>Genetic Vectors</term><term>Liposomes</term><term>Phosphatidylethanolamines</term><term>Plasmids</term><term>Quaternary Ammonium Compounds</term><term>Rats</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules cultivées</term><term>Cellules épithéliales</term><term>Composés d'ammonium quaternaire</term><term>Cytomegalovirus</term><term>Foetus</term><term>Gènes rapporteurs</term><term>Liposomes</term><term>Phosphatidyléthanolamine</term><term>Plasmides</term><term>Poumon</term><term>Rats</term><term>Survie cellulaire</term><term>Transfection</term><term>Vecteurs de médicaments</term><term>Vecteurs génétiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Cationic liposomes, 1:1 (mol/mol) 1,2-dioleoyldimethylammonium chloride-1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, were used to transfect primary cultures of distal rat fetal lung epithelial cells with pCMV4-based plasmids. A DNA-to-lipid ratio of 1:10 to 1:15 (wt/wt) optimized DNA uptake over a 24-h exposure. At a fixed DNA-to-lipid ratio of 1:15, chloramphenicol acetyltransferase (CAT) reporter gene expression declined at lipid concentrations > 2.5 nmol/cm2 cell surface area, whereas DNA uptake remained concentration dependent. CAT expression peaked 48 h after removal of the liposome-DNA complex, declining thereafter. Reporter gene expression was increased, and supercoiled cDNA degradation was reduced by the addition of 0.2 mM nicotinamide and 10 microM chloroquine. Rat fetal lung epithelial cells transfected with two different expression cassettes had an increased susceptibility to superoxide-mediated cytotoxicity. This could be attributed to a nonspecific delivery of exogenous DNA or some other copurified factor. The DNA-dependent increase in superoxide-mediated cytotoxicity, but not basal levels of cytotoxicity, was inhibited by the addition of 0.2 mM nicotinamide and 10 microM chloroquine.</div></front></TEI><affiliations><list><country><li>Canada</li></country><region><li>Ontario</li></region><settlement><li>Toronto</li></settlement><orgName><li>Université de Toronto</li></orgName></list><tree><noCountry><name sortKey="Belcastro, R" sort="Belcastro, R" uniqKey="Belcastro R" first="R" last="Belcastro">R. Belcastro</name><name sortKey="Cabacungan, J" sort="Cabacungan, J" uniqKey="Cabacungan J" first="J" last="Cabacungan">J. Cabacungan</name><name sortKey="Hu, J" sort="Hu, J" uniqKey="Hu J" first="J" last="Hu">J. Hu</name><name sortKey="Iles, R" sort="Iles, R" uniqKey="Iles R" first="R" last="Iles">R. Iles</name><name sortKey="O Brodovich, H" sort="O Brodovich, H" uniqKey="O Brodovich H" first="H" last="O'Brodovich">H. O'Brodovich</name><name sortKey="Sedlackova, L" sort="Sedlackova, L" uniqKey="Sedlackova L" first="L" last="Sedlackova">L. Sedlackova</name><name sortKey="Staub, O" sort="Staub, O" uniqKey="Staub O" first="O" last="Staub">O. Staub</name><name sortKey="Steer, B" sort="Steer, B" uniqKey="Steer B" first="B" last="Steer">B. Steer</name><name sortKey="Wen, Y" sort="Wen, Y" uniqKey="Wen Y" first="Y" last="Wen">Y. Wen</name></noCountry><country name="Canada"><region name="Ontario"><name sortKey="Tanswell, A K" sort="Tanswell, A K" uniqKey="Tanswell A" first="A K" last="Tanswell">A K Tanswell</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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