Liposome-mediated transfection of fetal lung epithelial cells : DNA degradation and enhanced superoxide toxicity
Identifieur interne : 002847 ( Main/Exploration ); précédent : 002846; suivant : 002848Liposome-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 ; YANXIA WEN ; J. Hu ; H. O'BrodovichSource :
- American journal of physiology. Lung cellular and molecular physiology [ 1040-0605 ] ; 1998.
Descripteurs français
- Pascal (Inist)
English descriptors
- KwdEn :
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 μM 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 μM chloroquine.
Affiliations:
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Tanswell</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>The Medical Research Council Group in Lung Development and Lung Biology Programme, Hospital for Sick Children Research Institute, University of Toronto</s1><s2>Toronto, Ontario, M5G 1X8</s2><s3>CAN</s3></inist:fA14><country>Canada</country><placeName><settlement type="city">Toronto</settlement><region type="state">Ontario</region></placeName><orgName type="university">Université de Toronto</orgName></affiliation><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Divisions of Neonatology and Respiratory Medicine, Department of Paediatrics, University of Toronto</s1><s2>Toronto, Ontario, M5G 1X8</s2><s3>CAN</s3></inist:fA14><country>Canada</country><placeName><settlement type="city">Toronto</settlement><region type="state">Ontario</region></placeName><orgName type="university">Université de Toronto</orgName></affiliation></author><author><name sortKey="Staub, O" sort="Staub, O" uniqKey="Staub O" first="O." last="Staub">O. Staub</name></author><author><name sortKey="Iles, R" sort="Iles, R" uniqKey="Iles R" first="R." last="Iles">R. Iles</name></author><author><name sortKey="Belcastro, R" sort="Belcastro, R" uniqKey="Belcastro R" first="R." last="Belcastro">R. Belcastro</name></author><author><name sortKey="Cabacungan, J" sort="Cabacungan, J" uniqKey="Cabacungan J" first="J." last="Cabacungan">J. Cabacungan</name></author><author><name sortKey="Sedlackova, L" sort="Sedlackova, L" uniqKey="Sedlackova L" first="L." last="Sedlackova">L. Sedlackova</name></author><author><name sortKey="Steer, B" sort="Steer, B" uniqKey="Steer B" first="B." last="Steer">B. Steer</name></author><author><name sortKey="Yanxia Wen" sort="Yanxia Wen" uniqKey="Yanxia Wen" last="Yanxia Wen">YANXIA WEN</name></author><author><name sortKey="Hu, J" sort="Hu, J" uniqKey="Hu J" first="J." last="Hu">J. Hu</name></author><author><name sortKey="O Brodovich, H" sort="O Brodovich, H" uniqKey="O Brodovich H" first="H." last="O'Brodovich">H. 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K." last="Tanswell">A. K. Tanswell</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>The Medical Research Council Group in Lung Development and Lung Biology Programme, Hospital for Sick Children Research Institute, University of Toronto</s1><s2>Toronto, Ontario, M5G 1X8</s2><s3>CAN</s3></inist:fA14><country>Canada</country><placeName><settlement type="city">Toronto</settlement><region type="state">Ontario</region></placeName><orgName type="university">Université de Toronto</orgName></affiliation><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Divisions of Neonatology and Respiratory Medicine, Department of Paediatrics, University of Toronto</s1><s2>Toronto, Ontario, M5G 1X8</s2><s3>CAN</s3></inist:fA14><country>Canada</country><placeName><settlement type="city">Toronto</settlement><region type="state">Ontario</region></placeName><orgName type="university">Université de Toronto</orgName></affiliation></author><author><name sortKey="Staub, O" sort="Staub, O" uniqKey="Staub O" first="O." last="Staub">O. Staub</name></author><author><name sortKey="Iles, R" sort="Iles, R" uniqKey="Iles R" first="R." last="Iles">R. Iles</name></author><author><name sortKey="Belcastro, R" sort="Belcastro, R" uniqKey="Belcastro R" first="R." last="Belcastro">R. Belcastro</name></author><author><name sortKey="Cabacungan, J" sort="Cabacungan, J" uniqKey="Cabacungan J" first="J." last="Cabacungan">J. Cabacungan</name></author><author><name sortKey="Sedlackova, L" sort="Sedlackova, L" uniqKey="Sedlackova L" first="L." last="Sedlackova">L. Sedlackova</name></author><author><name sortKey="Steer, B" sort="Steer, B" uniqKey="Steer B" first="B." last="Steer">B. Steer</name></author><author><name sortKey="Yanxia Wen" sort="Yanxia Wen" uniqKey="Yanxia Wen" last="Yanxia Wen">YANXIA WEN</name></author><author><name sortKey="Hu, J" sort="Hu, J" uniqKey="Hu J" first="J." last="Hu">J. Hu</name></author><author><name sortKey="O Brodovich, H" sort="O Brodovich, H" uniqKey="O Brodovich H" first="H." last="O'Brodovich">H. O'Brodovich</name></author></analytic><series><title level="j" type="main">American journal of physiology. Lung cellular and molecular physiology</title><title level="j" type="abbreviated">Am. j. physiol., Lung cell. mol. physiol.</title><idno type="ISSN">1040-0605</idno><imprint><date when="1998">1998</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">American journal of physiology. Lung cellular and molecular physiology</title><title level="j" type="abbreviated">Am. j. physiol., Lung cell. mol. physiol.</title><idno type="ISSN">1040-0605</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cytotoxicity</term><term>DNA</term><term>Epithelial cell</term><term>Fetus</term><term>Genetic transfer</term><term>In vitro</term><term>Liposome</term><term>Lung</term><term>Molecular interaction</term><term>Rat</term><term>Transfection</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Poumon</term><term>Foetus</term><term>Transfection</term><term>Liposome</term><term>Transfert génétique</term><term>DNA</term><term>Interaction moléculaire</term><term>Cytotoxicité</term><term>In vitro</term><term>Rat</term><term>Cellule épithéliale</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/cm<sup>2</sup> 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 μM 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 μM 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="Yanxia Wen" sort="Yanxia Wen" uniqKey="Yanxia Wen" last="Yanxia Wen">YANXIA 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><name sortKey="Tanswell, A K" sort="Tanswell, A K" uniqKey="Tanswell A" first="A. K." last="Tanswell">A. K. Tanswell</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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