Vitellogenin synthesis in cultured hepatocytes; an in vitro test for the estrogenic potency of chemicals
Identifieur interne : 001397 ( Istex/Corpus ); précédent : 001396; suivant : 001398Vitellogenin synthesis in cultured hepatocytes; an in vitro test for the estrogenic potency of chemicals
Auteurs : C. Pelissero ; G. Flouriot ; J. L. Foucher ; B. Bennetau ; J. Dunoguès ; F. Le Gac ; J. P. SumpterSource :
- Journal of Steroid Biochemistry and Molecular Biology [ 0960-0760 ] ; 1993.
Abstract
We describe here an in vitro technique to assess the estrogenic activity of chemicals. This technique is based on rainbow trout hepatocytes incubated in a basic medium free of any additional growth factors or estrogenic chemicals and uses the production of vitellogenin (VTG) as a marker for the estrogenic potency of the compounds tested. The system allows at least some of the metabolic transformations which are undertaken by the liver cells in vivo and could therefore be used for xenobiotic compounds which exhibit estrogenic activities after liver metabolic transformation. A dose-response curve was always consistently obtained using estradiol-17β (E2), with a mid point at around 100 nM E2 and a maximum response at around 1000 nM. Established estrogens such as 17 a 1 ethynylestradiol (EE2) or diethylstilboestrol (DES) were also tested. EE2 appeared to be equipotent with E2 and DES slightly less potent. E2 conjugates were, perhaps surprisingly, also very potent. Estradiol-3-sulfate was equipotent with E2 and estradiol-17β-glucuronide approx. 10% as potent. Other steroids such as androgens and progesterone, though active in the bioassay, were 3 orders of magnitude less potent than E2. Of the various steroids tested, only cortisol, at concentrations up to 50 μM, was completely inactive. Six different phytoestrogens were tested in the assay. All were weakly estrogenic, possessing approximately one thousanth the potency of E2 (they were as potent as the endrogens and progesterone). All six phytoestrogens, as well as the androgens and progesterone, were tested in the presence of tamoxifen. In all cases tamoxifen reduced the production of VTG significantly, demonstrating that the estrogenic action of all of these compounds was most likely mediated by the E2 receptor. The potencies determined here may not reflect the situation in vivo but can provide complementary results about the activity of chemicals which need an hepatic metabolization to be estrogenic. Hepatocyte cultures would profitably be developed in other species to sustain these results.
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DOI: 10.1016/0960-0760(93)90086-C
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Pelissero</name><affiliation><mods:affiliation>Department of Biology and Biochemistry, Brunel University, Uxbridge, Middlesex UB8 3PH, England</mods:affiliation></affiliation></author><author><name sortKey="Flouriot, G" sort="Flouriot, G" uniqKey="Flouriot G" first="G." last="Flouriot">G. Flouriot</name><affiliation><mods:affiliation>Laboratoire de Biologie Moléculaire Université de Rennes I, Campus de Beaulieu, 35042 Rennes Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Foucher, J L" sort="Foucher, J L" uniqKey="Foucher J" first="J. L." last="Foucher">J. L. Foucher</name><affiliation><mods:affiliation>Laboratoire INRA de Physiologie des Poissons, Campus de Rennes Beaulieu, 35042 Rennes Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Bennetau, B" sort="Bennetau, B" uniqKey="Bennetau B" first="B." last="Bennetau">B. Bennetau</name><affiliation><mods:affiliation>Laboratoire de Chimie Organique et Organométallique, URA 35 CNRS, Université de Bordeaux I, 351, Cours de la Libération, 33405 Talence Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Dunogues, J" sort="Dunogues, J" uniqKey="Dunogues J" first="J." last="Dunoguès">J. Dunoguès</name><affiliation><mods:affiliation>Laboratoire de Chimie Organique et Organométallique, URA 35 CNRS, Université de Bordeaux I, 351, Cours de la Libération, 33405 Talence Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Le Gac, F" sort="Le Gac, F" uniqKey="Le Gac F" first="F." last="Le Gac">F. Le Gac</name><affiliation><mods:affiliation>Laboratoire INRA de Physiologie des Poissons, Campus de Rennes Beaulieu, 35042 Rennes Cedex France</mods:affiliation></affiliation></author><author><name sortKey="Sumpter, J P" sort="Sumpter, J P" uniqKey="Sumpter J" first="J. P." last="Sumpter">J. P. Sumpter</name><affiliation><mods:affiliation>Department of Biology and Biochemistry, Brunel University, Uxbridge, Middlesex UB8 3PH, England</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Steroid Biochemistry and Molecular Biology</title><title level="j" type="abbrev">SBMB</title><idno type="ISSN">0960-0760</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1993">1993</date><biblScope unit="volume">44</biblScope><biblScope unit="issue">3</biblScope><biblScope unit="page" from="263">263</biblScope><biblScope unit="page" to="272">272</biblScope></imprint><idno type="ISSN">0960-0760</idno></series><idno type="istex">BC82C0D67A25B0683FF26F965B03366DA4BFD76E</idno><idno type="DOI">10.1016/0960-0760(93)90086-C</idno><idno type="PII">0960-0760(93)90086-C</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0960-0760</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">We describe here an in vitro technique to assess the estrogenic activity of chemicals. This technique is based on rainbow trout hepatocytes incubated in a basic medium free of any additional growth factors or estrogenic chemicals and uses the production of vitellogenin (VTG) as a marker for the estrogenic potency of the compounds tested. The system allows at least some of the metabolic transformations which are undertaken by the liver cells in vivo and could therefore be used for xenobiotic compounds which exhibit estrogenic activities after liver metabolic transformation. A dose-response curve was always consistently obtained using estradiol-17β (E2), with a mid point at around 100 nM E2 and a maximum response at around 1000 nM. Established estrogens such as 17 a 1 ethynylestradiol (EE2) or diethylstilboestrol (DES) were also tested. EE2 appeared to be equipotent with E2 and DES slightly less potent. E2 conjugates were, perhaps surprisingly, also very potent. Estradiol-3-sulfate was equipotent with E2 and estradiol-17β-glucuronide approx. 10% as potent. Other steroids such as androgens and progesterone, though active in the bioassay, were 3 orders of magnitude less potent than E2. Of the various steroids tested, only cortisol, at concentrations up to 50 μM, was completely inactive. Six different phytoestrogens were tested in the assay. All were weakly estrogenic, possessing approximately one thousanth the potency of E2 (they were as potent as the endrogens and progesterone). All six phytoestrogens, as well as the androgens and progesterone, were tested in the presence of tamoxifen. In all cases tamoxifen reduced the production of VTG significantly, demonstrating that the estrogenic action of all of these compounds was most likely mediated by the E2 receptor. The potencies determined here may not reflect the situation in vivo but can provide complementary results about the activity of chemicals which need an hepatic metabolization to be estrogenic. Hepatocyte cultures would profitably be developed in other species to sustain these results.</div></front></TEI><istex><corpusName>elsevier</corpusName><author><json:item><name>C. Pelissero</name><affiliations><json:string>Department of Biology and Biochemistry, Brunel University, Uxbridge, Middlesex UB8 3PH, England</json:string></affiliations></json:item><json:item><name>G. Flouriot</name><affiliations><json:string>Laboratoire de Biologie Moléculaire Université de Rennes I, Campus de Beaulieu, 35042 Rennes Cedex France</json:string></affiliations></json:item><json:item><name>J.L. Foucher</name><affiliations><json:string>Laboratoire INRA de Physiologie des Poissons, Campus de Rennes Beaulieu, 35042 Rennes Cedex France</json:string></affiliations></json:item><json:item><name>B. 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A dose-response curve was always consistently obtained using estradiol-17β (E2), with a mid point at around 100 nM E2 and a maximum response at around 1000 nM. Established estrogens such as 17 a 1 ethynylestradiol (EE2) or diethylstilboestrol (DES) were also tested. EE2 appeared to be equipotent with E2 and DES slightly less potent. E2 conjugates were, perhaps surprisingly, also very potent. Estradiol-3-sulfate was equipotent with E2 and estradiol-17β-glucuronide approx. 10% as potent. Other steroids such as androgens and progesterone, though active in the bioassay, were 3 orders of magnitude less potent than E2. Of the various steroids tested, only cortisol, at concentrations up to 50 μM, was completely inactive. Six different phytoestrogens were tested in the assay. All were weakly estrogenic, possessing approximately one thousanth the potency of E2 (they were as potent as the endrogens and progesterone). All six phytoestrogens, as well as the androgens and progesterone, were tested in the presence of tamoxifen. In all cases tamoxifen reduced the production of VTG significantly, demonstrating that the estrogenic action of all of these compounds was most likely mediated by the E2 receptor. The potencies determined here may not reflect the situation in vivo but can provide complementary results about the activity of chemicals which need an hepatic metabolization to be estrogenic. 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This technique is based on rainbow trout hepatocytes incubated in a basic medium free of any additional growth factors or estrogenic chemicals and uses the production of vitellogenin (VTG) as a marker for the estrogenic potency of the compounds tested. The system allows at least some of the metabolic transformations which are undertaken by the liver cells in vivo and could therefore be used for xenobiotic compounds which exhibit estrogenic activities after liver metabolic transformation. A dose-response curve was always consistently obtained using estradiol-17β (E2), with a mid point at around 100 nM E2 and a maximum response at around 1000 nM. Established estrogens such as 17 a 1 ethynylestradiol (EE2) or diethylstilboestrol (DES) were also tested. EE2 appeared to be equipotent with E2 and DES slightly less potent. E2 conjugates were, perhaps surprisingly, also very potent. Estradiol-3-sulfate was equipotent with E2 and estradiol-17β-glucuronide approx. 10% as potent. Other steroids such as androgens and progesterone, though active in the bioassay, were 3 orders of magnitude less potent than E2. Of the various steroids tested, only cortisol, at concentrations up to 50 μM, was completely inactive. Six different phytoestrogens were tested in the assay. All were weakly estrogenic, possessing approximately one thousanth the potency of E2 (they were as potent as the endrogens and progesterone). All six phytoestrogens, as well as the androgens and progesterone, were tested in the presence of tamoxifen. In all cases tamoxifen reduced the production of VTG significantly, demonstrating that the estrogenic action of all of these compounds was most likely mediated by the E2 receptor. The potencies determined here may not reflect the situation in vivo but can provide complementary results about the activity of chemicals which need an hepatic metabolization to be estrogenic. Hepatocyte cultures would profitably be developed in other species to sustain these results.</p></abstract></profileDesc><revisionDesc><change when="1993">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/document/BC82C0D67A25B0683FF26F965B03366DA4BFD76E/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: tail"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla"><item-info><jid>SBMB</jid><aid>9390086C</aid><ce:pii>0960-0760(93)90086-C</ce:pii><ce:doi>10.1016/0960-0760(93)90086-C</ce:doi><ce:copyright type="unknown" year="1993"></ce:copyright></item-info><head><ce:title>Vitellogenin synthesis in cultured hepatocytes; an <ce:italic>in vitro</ce:italic> test for the estrogenic potency of chemicals</ce:title><ce:author-group><ce:author><ce:given-name>C.</ce:given-name><ce:surname>Pelissero</ce:surname><ce:cross-ref refid="COR1"><ce:sup>∗</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF1"><ce:sup>1</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>G.</ce:given-name><ce:surname>Flouriot</ce:surname><ce:cross-ref refid="AFF2"><ce:sup>2</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>J.L.</ce:given-name><ce:surname>Foucher</ce:surname><ce:cross-ref refid="AFF3"><ce:sup>3</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>B.</ce:given-name><ce:surname>Bennetau</ce:surname><ce:cross-ref refid="AFF4"><ce:sup>4</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>J.</ce:given-name><ce:surname>Dunoguès</ce:surname><ce:cross-ref refid="AFF4"><ce:sup>4</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>F.</ce:given-name><ce:surname>Le Gac</ce:surname><ce:cross-ref refid="AFF3"><ce:sup>3</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>J.P.</ce:given-name><ce:surname>Sumpter</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>1</ce:sup></ce:cross-ref></ce:author><ce:affiliation id="AFF1"><ce:label>1</ce:label><ce:textfn>Department of Biology and Biochemistry, Brunel University, Uxbridge, Middlesex UB8 3PH, England</ce:textfn></ce:affiliation><ce:affiliation id="AFF2"><ce:label>2</ce:label><ce:textfn>Laboratoire de Biologie Moléculaire Université de Rennes I, Campus de Beaulieu, 35042 Rennes Cedex France</ce:textfn></ce:affiliation><ce:affiliation id="AFF3"><ce:label>3</ce:label><ce:textfn>Laboratoire INRA de Physiologie des Poissons, Campus de Rennes Beaulieu, 35042 Rennes Cedex France</ce:textfn></ce:affiliation><ce:affiliation id="AFF4"><ce:label>4</ce:label><ce:textfn>Laboratoire de Chimie Organique et Organométallique, URA 35 CNRS, Université de Bordeaux I, 351, Cours de la Libération, 33405 Talence Cedex France</ce:textfn></ce:affiliation><ce:correspondence id="COR1"><ce:label>∗</ce:label><ce:text>To whom correspondence should be addressed at his present address: Laboratoire d'Océanographie Biologique, Université de Bordeaux I, Avenue des Facultés, 33405 Talence Cédex, France.</ce:text></ce:correspondence></ce:author-group><ce:date-received day="28" month="7" year="1992"></ce:date-received><ce:date-accepted day="9" month="11" year="1992"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>We describe here an <ce:italic>in vitro</ce:italic> technique to assess the estrogenic activity of chemicals. This technique is based on rainbow trout hepatocytes incubated in a basic medium free of any additional growth factors or estrogenic chemicals and uses the production of vitellogenin (VTG) as a marker for the estrogenic potency of the compounds tested. The system allows at least some of the metabolic transformations which are undertaken by the liver cells <ce:italic>in vivo</ce:italic> and could therefore be used for xenobiotic compounds which exhibit estrogenic activities after liver metabolic transformation. A dose-response curve was always consistently obtained using estradiol-17β (E<ce:inf>2</ce:inf>), with a mid point at around 100 nM E<ce:inf>2</ce:inf> and a maximum response at around 1000 nM. Established estrogens such as 17 <ce:italic>a</ce:italic> 1 ethynylestradiol (EE<ce:inf>2</ce:inf>) or diethylstilboestrol (DES) were also tested. EE<ce:inf>2</ce:inf> appeared to be equipotent with E<ce:inf>2</ce:inf> and DES slightly less potent. E<ce:inf>2</ce:inf> conjugates were, perhaps surprisingly, also very potent. Estradiol-3-sulfate was equipotent with E<ce:inf>2</ce:inf> and estradiol-17β-glucuronide approx. 10% as potent. Other steroids such as androgens and progesterone, though active in the bioassay, were 3 orders of magnitude less potent than E<ce:inf>2</ce:inf>. Of the various steroids tested, only cortisol, at concentrations up to 50 μM, was completely inactive. Six different phytoestrogens were tested in the assay. All were weakly estrogenic, possessing approximately one thousanth the potency of E<ce:inf>2</ce:inf> (they were as potent as the endrogens and progesterone). All six phytoestrogens, as well as the androgens and progesterone, were tested in the presence of tamoxifen. In all cases tamoxifen reduced the production of VTG significantly, demonstrating that the estrogenic action of all of these compounds was most likely mediated by the E<ce:inf>2</ce:inf> receptor. The potencies determined here may not reflect the situation <ce:italic>in vivo</ce:italic> but can provide complementary results about the activity of chemicals which need an hepatic metabolization to be estrogenic. Hepatocyte cultures would profitably be developed in other species to sustain these results.</ce:simple-para></ce:abstract-sec></ce:abstract></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Vitellogenin synthesis in cultured hepatocytes; an in vitro test for the estrogenic potency of chemicals</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Vitellogenin synthesis in cultured hepatocytes; an</title></titleInfo><name type="personal"><namePart type="given">C.</namePart><namePart type="family">Pelissero</namePart><affiliation>Department of Biology and Biochemistry, Brunel University, Uxbridge, Middlesex UB8 3PH, England</affiliation><description>To whom correspondence should be addressed at his present address: Laboratoire d'Océanographie Biologique, Université de Bordeaux I, Avenue des Facultés, 33405 Talence Cédex, France.</description><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">G.</namePart><namePart type="family">Flouriot</namePart><affiliation>Laboratoire de Biologie Moléculaire Université de Rennes I, Campus de Beaulieu, 35042 Rennes Cedex France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">J.L.</namePart><namePart type="family">Foucher</namePart><affiliation>Laboratoire INRA de Physiologie des Poissons, Campus de Rennes Beaulieu, 35042 Rennes Cedex France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">B.</namePart><namePart type="family">Bennetau</namePart><affiliation>Laboratoire de Chimie Organique et Organométallique, URA 35 CNRS, Université de Bordeaux I, 351, Cours de la Libération, 33405 Talence Cedex France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">J.</namePart><namePart type="family">Dunoguès</namePart><affiliation>Laboratoire de Chimie Organique et Organométallique, URA 35 CNRS, Université de Bordeaux I, 351, Cours de la Libération, 33405 Talence Cedex France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">F.</namePart><namePart type="family">Le Gac</namePart><affiliation>Laboratoire INRA de Physiologie des Poissons, Campus de Rennes Beaulieu, 35042 Rennes Cedex France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">J.P.</namePart><namePart type="family">Sumpter</namePart><affiliation>Department of Biology and Biochemistry, Brunel University, Uxbridge, Middlesex UB8 3PH, England</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article"></genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1993</dateIssued><copyrightDate encoding="w3cdtf">1993</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract lang="en">We describe here an in vitro technique to assess the estrogenic activity of chemicals. This technique is based on rainbow trout hepatocytes incubated in a basic medium free of any additional growth factors or estrogenic chemicals and uses the production of vitellogenin (VTG) as a marker for the estrogenic potency of the compounds tested. The system allows at least some of the metabolic transformations which are undertaken by the liver cells in vivo and could therefore be used for xenobiotic compounds which exhibit estrogenic activities after liver metabolic transformation. A dose-response curve was always consistently obtained using estradiol-17β (E2), with a mid point at around 100 nM E2 and a maximum response at around 1000 nM. Established estrogens such as 17 a 1 ethynylestradiol (EE2) or diethylstilboestrol (DES) were also tested. EE2 appeared to be equipotent with E2 and DES slightly less potent. E2 conjugates were, perhaps surprisingly, also very potent. Estradiol-3-sulfate was equipotent with E2 and estradiol-17β-glucuronide approx. 10% as potent. Other steroids such as androgens and progesterone, though active in the bioassay, were 3 orders of magnitude less potent than E2. Of the various steroids tested, only cortisol, at concentrations up to 50 μM, was completely inactive. Six different phytoestrogens were tested in the assay. All were weakly estrogenic, possessing approximately one thousanth the potency of E2 (they were as potent as the endrogens and progesterone). All six phytoestrogens, as well as the androgens and progesterone, were tested in the presence of tamoxifen. In all cases tamoxifen reduced the production of VTG significantly, demonstrating that the estrogenic action of all of these compounds was most likely mediated by the E2 receptor. The potencies determined here may not reflect the situation in vivo but can provide complementary results about the activity of chemicals which need an hepatic metabolization to be estrogenic. Hepatocyte cultures would profitably be developed in other species to sustain these results.</abstract><relatedItem type="host"><titleInfo><title>Journal of Steroid Biochemistry and Molecular Biology</title></titleInfo><titleInfo type="abbreviated"><title>SBMB</title></titleInfo><genre type="journal">journal</genre><originInfo><dateIssued encoding="w3cdtf">199303</dateIssued></originInfo><identifier type="ISSN">0960-0760</identifier><identifier type="PII">S0960-0760(00)X0107-0</identifier><part><date>199303</date><detail type="volume"><number>44</number><caption>vol.</caption></detail><detail type="issue"><number>3</number><caption>no.</caption></detail><extent unit="issue pages"><start>203</start><end>320</end></extent><extent unit="pages"><start>263</start><end>272</end></extent></part></relatedItem><identifier type="istex">BC82C0D67A25B0683FF26F965B03366DA4BFD76E</identifier><identifier type="DOI">10.1016/0960-0760(93)90086-C</identifier><identifier type="PII">0960-0760(93)90086-C</identifier><recordInfo><recordContentSource>ELSEVIER</recordContentSource></recordInfo></mods></metadata></istex></record>Pour manipuler ce document sous Unix (Dilib)
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