Putative Role of Chloroquine in Gene Transfer into a Human Hepatoma Cell Line by DNA/Lactosylated Polylysine Complexes
Identifieur interne : 002968 ( Main/Exploration ); précédent : 002967; suivant : 002969Putative Role of Chloroquine in Gene Transfer into a Human Hepatoma Cell Line by DNA/Lactosylated Polylysine Complexes
Auteurs : Patrick Erbacher [France] ; Annie Claude Roche [France] ; Michel Monsigny [France] ; Patrick Midoux [France]Source :
- Experimental Cell Research [ 0014-4827 ] ; 1996-05.
Abstract
Chloroquine improves drastically the transfection of cells upon exposure to plasmid DNA/glycosylated polylysine complexes. So far the mechanism of action of chloroquine is not well understood. In this paper, the effect of chloroquine was investigated by measuring the transfection efficiency of a human hepatocarcinoma (HepG2 cells) by pSV2LUC/lactosylated polylysine complexes involving their internalization via the galactose-specific membrane lectin of these cells. The luciferase activity in the transfected cells was maximal when the transfection was performed for 3 or 4 h in the presence of 100 microM chloroquine. The luciferase activity was also enhanced in the presence of primaquine, a chloroquine analogue, but was not increased when transfection was performed in the presence of ammonium chloride, methylamine, spermine, or monensin, compounds known to neutralize the pH of the endocytotic vesicle lumen as chloroquine does. Chloroquine enters cells and accumulates in vesicular compartments; the overall intracellular concentration increases to 9 mM, which means that in the vesicular compartment, the chloroquine concentration is still higher. At such high concentrations, chloroquine induces the dissociation of plasmid DNA/lactosylated polylysine complexes, as shown in acellular experiments.
Url:
DOI: 10.1006/excr.1996.0169
Affiliations:
- France
- Centre-Val de Loire, Région Centre
- Orléans
- Centre Val de Loire Université, Université d'Orléans
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Le document en format XML
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<desc> <address> <addrLine>101, rue de Tolbiac, 75013 Paris </addrLine>
<country key="FR"></country>
</address>
<ref type="url">http://www.inserm.fr</ref>
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<idno type="ISNI">0000000122597504</idno>
<orgName>Centre National de la Recherche Scientifique</orgName>
<orgName type="acronym">CNRS</orgName>
<date type="start">1939-10-19</date>
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</address>
<ref type="url">http://www.cnrs.fr/</ref>
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<country>France</country>
<placeName><settlement type="city">Orléans</settlement>
<region type="old region" nuts="2">Région Centre</region>
<region type="region" nuts="2">Centre-Val de Loire</region>
</placeName>
<orgName type="university">Université d'Orléans</orgName>
<orgName type="institution" wicri:auto="newGroup">Centre Val de Loire Université</orgName>
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</author>
<author><name sortKey="Midoux, Patrick" sort="Midoux, Patrick" uniqKey="Midoux P" first="Patrick" last="Midoux">Patrick Midoux</name>
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<idno type="RNSR">196717612S</idno>
<orgName>Centre de biophysique moléculaire</orgName>
<orgName type="acronym">CBM</orgName>
<date type="start">1967-01-01</date>
<desc> <address> <addrLine>Rue Charles Sadron 45071 ORLEANS CEDEX 2</addrLine>
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</address>
<ref type="url">http://cbm.cnrs-orleans.fr/</ref>
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<orgName>Université d'Orléans</orgName>
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<desc> <address> <addrLine>Château de la Source - Avenue du Parc Floral - BP 6749 - 45067 Orléans cedex 2</addrLine>
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</address>
<ref type="url">http://www.univ-orleans.fr/</ref>
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<tutelle name="UPR4301" active="#struct-303623" type="direct"><org type="institution" xml:id="struct-303623" status="VALID"> <idno type="IdRef">026388278</idno>
<orgName>Institut National de la Santé et de la Recherche Médicale</orgName>
<orgName type="acronym">INSERM</orgName>
<desc> <address> <addrLine>101, rue de Tolbiac, 75013 Paris </addrLine>
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<idno type="ISNI">0000000122597504</idno>
<orgName>Centre National de la Recherche Scientifique</orgName>
<orgName type="acronym">CNRS</orgName>
<date type="start">1939-10-19</date>
<desc> <address> <country key="FR"></country>
</address>
<ref type="url">http://www.cnrs.fr/</ref>
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</hal:affiliation>
<country>France</country>
<placeName><settlement type="city">Orléans</settlement>
<region type="old region" nuts="2">Région Centre</region>
<region type="region" nuts="2">Centre-Val de Loire</region>
</placeName>
<orgName type="university">Université d'Orléans</orgName>
<orgName type="institution" wicri:auto="newGroup">Centre Val de Loire Université</orgName>
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<idno type="DOI">10.1006/excr.1996.0169</idno>
<series><title level="j">Experimental Cell Research</title>
<idno type="ISSN">0014-4827</idno>
<imprint><date type="datePub">1996-05</date>
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<front><div type="abstract" xml:lang="en"> <p>Chloroquine improves drastically the transfection of cells upon exposure to plasmid DNA/glycosylated polylysine complexes. So far the mechanism of action of chloroquine is not well understood. In this paper, the effect of chloroquine was investigated by measuring the transfection efficiency of a human hepatocarcinoma (HepG2 cells) by pSV2LUC/lactosylated polylysine complexes involving their internalization via the galactose-specific membrane lectin of these cells. The luciferase activity in the transfected cells was maximal when the transfection was performed for 3 or 4 h in the presence of 100 microM chloroquine. The luciferase activity was also enhanced in the presence of primaquine, a chloroquine analogue, but was not increased when transfection was performed in the presence of ammonium chloride, methylamine, spermine, or monensin, compounds known to neutralize the pH of the endocytotic vesicle lumen as chloroquine does. Chloroquine enters cells and accumulates in vesicular compartments; the overall intracellular concentration increases to 9 mM, which means that in the vesicular compartment, the chloroquine concentration is still higher. At such high concentrations, chloroquine induces the dissociation of plasmid DNA/lactosylated polylysine complexes, as shown in acellular experiments.</p>
</div>
</front>
</TEI>
<affiliations><list><country><li>France</li>
</country>
<region><li>Centre-Val de Loire</li>
<li>Région Centre</li>
</region>
<settlement><li>Orléans</li>
</settlement>
<orgName><li>Centre Val de Loire Université</li>
<li>Université d'Orléans</li>
</orgName>
</list>
<tree><country name="France"><region name="Région Centre"><name sortKey="Erbacher, Patrick" sort="Erbacher, Patrick" uniqKey="Erbacher P" first="Patrick" last="Erbacher">Patrick Erbacher</name>
</region>
<name sortKey="Midoux, Patrick" sort="Midoux, Patrick" uniqKey="Midoux P" first="Patrick" last="Midoux">Patrick Midoux</name>
<name sortKey="Monsigny, Michel" sort="Monsigny, Michel" uniqKey="Monsigny M" first="Michel" last="Monsigny">Michel Monsigny</name>
<name sortKey="Roche, Annie Claude" sort="Roche, Annie Claude" uniqKey="Roche A" first="Annie Claude" last="Roche">Annie Claude Roche</name>
</country>
</tree>
</affiliations>
</record>
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