Retinoic acid negatively regulates neuropeptide Y expression in human neuroblastoma cells
Identifieur interne : 000212 ( Main/Corpus ); précédent : 000211; suivant : 000213Retinoic acid negatively regulates neuropeptide Y expression in human neuroblastoma cells
Auteurs : Paolo Magni ; Elena Beretta ; Eugenia Scaccianoce ; Marcella MottaSource :
- Neuropharmacology [ 0028-3908 ] ; 1999.
Abstract
Retinoids are involved in the regulation of development and differentiation in many tissues, including the nervous system, where they have been associated with some neurotransmitter systems. In the present study, we evaluated the effects of all-trans retinoic acid (RA) on the biosynthesis and secretion of neuropeptide Y (NPY), a widely expressed neuroregulatory peptide. The SH-SY5Y human neuroblastoma cell line has been used as the in vitro model system. Treatment with 10 μM RA induced a marked decrease in NPY gene expression after as little as 3–6 h of incubation and resulted in its almost complete suppression at 12–24 h and after a 6-day differentiating treatment. The NPY content in cell extracts and the NPY secreted and accumulated in the culture medium were also reduced by exposure to 10 μM RA at 12 and 24 h and at 6 days. Moreover, RA treatment for 6 days, but not for 24 h, resulted in a marked stimulation of proNPY processing to mature NPY. The presence of negative retinoic acid-response elements in the human NPY promoter (up to −1078 bp) was excluded by a computer search. When SH-SY5Y cells were treated simultaneously with 20 nM TPA and 10 μM RA for 24 h, the marked stimulatory effect of TPA alone was completely suppressed. These observations suggest that the expression of NPY in SH-SY5Y human neuroblastoma cells is negatively regulated by RA at the level of gene expression, probably by mechanisms involving the interaction of activated RARs with transcription factors (such as AP-1).
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DOI: 10.1016/S0028-3908(99)00231-2
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<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>Retinoic acid negatively regulates neuropeptide Y expression in human neuroblastoma cells</title><author><name sortKey="Magni, Paolo" sort="Magni, Paolo" uniqKey="Magni P" first="Paolo" last="Magni">Paolo Magni</name><affiliation><mods:affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: paolo.magni@unimi.it</mods:affiliation></affiliation></author><author><name sortKey="Beretta, Elena" sort="Beretta, Elena" uniqKey="Beretta E" first="Elena" last="Beretta">Elena Beretta</name><affiliation><mods:affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</mods:affiliation></affiliation></author><author><name sortKey="Scaccianoce, Eugenia" sort="Scaccianoce, Eugenia" uniqKey="Scaccianoce E" first="Eugenia" last="Scaccianoce">Eugenia Scaccianoce</name><affiliation><mods:affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</mods:affiliation></affiliation></author><author><name sortKey="Motta, Marcella" sort="Motta, Marcella" uniqKey="Motta M" first="Marcella" last="Motta">Marcella Motta</name><affiliation><mods:affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:75363D06AF5372F57E111CEA14B2CFAE018A838A</idno><date when="2000" year="2000">2000</date><idno type="doi">10.1016/S0028-3908(99)00231-2</idno><idno type="url">https://api.istex.fr/document/75363D06AF5372F57E111CEA14B2CFAE018A838A/fulltext/pdf</idno><idno type="wicri:Area/Main/Corpus">000212</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a">Retinoic acid negatively regulates neuropeptide Y expression in human neuroblastoma cells</title><author><name sortKey="Magni, Paolo" sort="Magni, Paolo" uniqKey="Magni P" first="Paolo" last="Magni">Paolo Magni</name><affiliation><mods:affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: paolo.magni@unimi.it</mods:affiliation></affiliation></author><author><name sortKey="Beretta, Elena" sort="Beretta, Elena" uniqKey="Beretta E" first="Elena" last="Beretta">Elena Beretta</name><affiliation><mods:affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</mods:affiliation></affiliation></author><author><name sortKey="Scaccianoce, Eugenia" sort="Scaccianoce, Eugenia" uniqKey="Scaccianoce E" first="Eugenia" last="Scaccianoce">Eugenia Scaccianoce</name><affiliation><mods:affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</mods:affiliation></affiliation></author><author><name sortKey="Motta, Marcella" sort="Motta, Marcella" uniqKey="Motta M" first="Marcella" last="Motta">Marcella Motta</name><affiliation><mods:affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Neuropharmacology</title><title level="j" type="abbrev">NP</title><idno type="ISSN">0028-3908</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1999">1999</date><biblScope unit="volume">39</biblScope><biblScope unit="issue">9</biblScope><biblScope unit="page" from="1628">1628</biblScope><biblScope unit="page" to="1636">1636</biblScope></imprint><idno type="ISSN">0028-3908</idno></series><idno type="istex">75363D06AF5372F57E111CEA14B2CFAE018A838A</idno><idno type="DOI">10.1016/S0028-3908(99)00231-2</idno><idno type="PII">S0028-3908(99)00231-2</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0028-3908</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Retinoids are involved in the regulation of development and differentiation in many tissues, including the nervous system, where they have been associated with some neurotransmitter systems. In the present study, we evaluated the effects of all-trans retinoic acid (RA) on the biosynthesis and secretion of neuropeptide Y (NPY), a widely expressed neuroregulatory peptide. The SH-SY5Y human neuroblastoma cell line has been used as the in vitro model system. Treatment with 10 μM RA induced a marked decrease in NPY gene expression after as little as 3–6 h of incubation and resulted in its almost complete suppression at 12–24 h and after a 6-day differentiating treatment. The NPY content in cell extracts and the NPY secreted and accumulated in the culture medium were also reduced by exposure to 10 μM RA at 12 and 24 h and at 6 days. Moreover, RA treatment for 6 days, but not for 24 h, resulted in a marked stimulation of proNPY processing to mature NPY. The presence of negative retinoic acid-response elements in the human NPY promoter (up to −1078 bp) was excluded by a computer search. When SH-SY5Y cells were treated simultaneously with 20 nM TPA and 10 μM RA for 24 h, the marked stimulatory effect of TPA alone was completely suppressed. These observations suggest that the expression of NPY in SH-SY5Y human neuroblastoma cells is negatively regulated by RA at the level of gene expression, probably by mechanisms involving the interaction of activated RARs with transcription factors (such as AP-1).</div></front></TEI><istex><corpusName>elsevier</corpusName><author><json:item><name>Paolo Magni</name><affiliations><json:string>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</json:string><json:string>E-mail: paolo.magni@unimi.it</json:string></affiliations></json:item><json:item><name>Elena Beretta</name><affiliations><json:string>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</json:string></affiliations></json:item><json:item><name>Eugenia Scaccianoce</name><affiliations><json:string>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</json:string></affiliations></json:item><json:item><name>Marcella Motta</name><affiliations><json:string>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>Retinoic acid</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Neuropeptide Y gene expression</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Neuropeptide Y biosynthesis</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Neuroblastoma</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Cell differentiation</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Phorbol ester</value></json:item></subject><language><json:string>eng</json:string></language><abstract>Retinoids are involved in the regulation of development and differentiation in many tissues, including the nervous system, where they have been associated with some neurotransmitter systems. In the present study, we evaluated the effects of all-trans retinoic acid (RA) on the biosynthesis and secretion of neuropeptide Y (NPY), a widely expressed neuroregulatory peptide. The SH-SY5Y human neuroblastoma cell line has been used as the in vitro model system. Treatment with 10 μM RA induced a marked decrease in NPY gene expression after as little as 3–6 h of incubation and resulted in its almost complete suppression at 12–24 h and after a 6-day differentiating treatment. The NPY content in cell extracts and the NPY secreted and accumulated in the culture medium were also reduced by exposure to 10 μM RA at 12 and 24 h and at 6 days. Moreover, RA treatment for 6 days, but not for 24 h, resulted in a marked stimulation of proNPY processing to mature NPY. The presence of negative retinoic acid-response elements in the human NPY promoter (up to −1078 bp) was excluded by a computer search. When SH-SY5Y cells were treated simultaneously with 20 nM TPA and 10 μM RA for 24 h, the marked stimulatory effect of TPA alone was completely suppressed. These observations suggest that the expression of NPY in SH-SY5Y human neuroblastoma cells is negatively regulated by RA at the level of gene expression, probably by mechanisms involving the interaction of activated RARs with transcription factors (such as AP-1).</abstract><qualityIndicators><score>7.898</score><pdfVersion>1.2</pdfVersion><pdfPageSize>598 x 795 pts</pdfPageSize><refBibsNative>true</refBibsNative><keywordCount>6</keywordCount><abstractCharCount>1514</abstractCharCount><pdfWordCount>4910</pdfWordCount><pdfCharCount>29866</pdfCharCount><pdfPageCount>9</pdfPageCount><abstractWordCount>249</abstractWordCount></qualityIndicators><title>Retinoic acid negatively regulates neuropeptide Y expression in human neuroblastoma cells</title><pii><json:string>S0028-3908(99)00231-2</json:string></pii><genre><json:string>research-article</json:string></genre><host><volume>39</volume><pii><json:string>S0028-3908(00)X0061-5</json:string></pii><pages><last>1636</last><first>1628</first></pages><issn><json:string>0028-3908</json:string></issn><issue>9</issue><genre></genre><language><json:string>unknown</json:string></language><title>Neuropharmacology</title><publicationDate>2000</publicationDate></host><categories><wos><json:string>PHARMACOLOGY & PHARMACY</json:string><json:string>NEUROSCIENCES</json:string></wos></categories><publicationDate>1999</publicationDate><copyrightDate>2000</copyrightDate><doi><json:string>10.1016/S0028-3908(99)00231-2</json:string></doi><id>75363D06AF5372F57E111CEA14B2CFAE018A838A</id><fulltext><json:item><original>true</original><mimetype>application/pdf</mimetype><extension>pdf</extension><uri>https://api.istex.fr/document/75363D06AF5372F57E111CEA14B2CFAE018A838A/fulltext/pdf</uri></json:item><json:item><original>false</original><mimetype>application/zip</mimetype><extension>zip</extension><uri>https://api.istex.fr/document/75363D06AF5372F57E111CEA14B2CFAE018A838A/fulltext/zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/document/75363D06AF5372F57E111CEA14B2CFAE018A838A/fulltext/tei"><teiHeader><fileDesc><titleStmt><title level="a">Retinoic acid negatively regulates neuropeptide Y expression in human neuroblastoma cells</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>ELSEVIER</publisher><availability><p>ELSEVIER</p></availability><date>2000</date></publicationStmt><notesStmt><note type="content">Fig. 1: Effect of treatment with RA on NPY gene expression (Northern blot analysis). Cells were treated for 3, 6, 12, 24 h and six days with 10 μM RA. Total RNA (30 μg/lane) was analyzed by Northern blot hybridization (C=control; RA=all-trans retinoic acid). The stripped blots were rehybridized for GAPDH mRNA for normalization. One Northern blot, representative of three separate experiments, is shown.</note><note type="content">Fig. 2: Effect of the combined treatment with RA and TPA on NPY gene expression (Northern blot analysis). Cells were treated for 24 h with RA at different concentrations, in the presence of 20 nM TPA, or in its absence. Total RNA (30 μg/lane) was analyzed by Northern blot hybridization (C=control; RA=all-trans retinoic acid). The stripped blots were rehybridized for GAPDH mRNA for normalization. One Northern blot, representative of three separate experiments, is shown.</note><note type="content">Fig. 3: Morphological changes of SH-SY5Y human neuroblastoma cells after differentiation with RA. Cells were treated for six days with 10 μM RA (all-trans retinoic acid) or solvent (C=control). The effect of treatment with 20 nM TPA for six days is shown for comparison.</note><note type="content">Fig. 4: Effect of treatment with RA on NPY synthesis/secretion. Cells were treated for 3, 6, 12, 24 h and six days with 10 μM RA and the NPY-IR content in cell extracts and in the medium was quantified by RIA. Panel A: effect of RA treatment on NPY-IR content in cell extracts. Panel B: effect of RA treatment on NPY-IR content in the medium. One experiment representative of three or four separate experiments is shown. Data are expressed as mean±S.E.M. (percent of controls=100). C=control; RA=all-trans retinoic acid; n=6; *P<0.05 vs C; **P<0.01 vs C; ***P<0.001 vs C (ANOVA).</note><note type="content">Fig. 5: Effect of 24-h and 6-day treatments with 10 μM RA on NPY precursor processing (cell extracts and conditioned medium). One experiment representative of 3–5 separate experiments is shown. Cell extracts (panel A) and conditioned media (panel B) were size-fractionated by gel filtration chromatography and the NPY-IR in each fraction was quantified by RIA. Dotted line: C (control); solid line: RA (all-trans retinoic acid). Please note that, for the purpose of clarity, the elution profiles of samples from cells treated for 24 h and for six days are separately superimposed to the same control elution profile (dotted line). In the upper left and in the lower right profiles is indicated the elution position of synthetic human NPY (“NPY std”), used as the reference standard, and the immunoreactive peak corresponding to the proNPY-like substance (“proNPY-like”). The table under each panel summarizes the data of gel filtration chromatography obtained in all the experiments performed and includes the representative chromatograms shown above them. The data (mean±S.E.M.) relative to the proNPY-like and to the NPY peaks are expressed as % of total NPY-IR (total NPY-IR=sum of the areas under the curve relative to the two chromatographic peaks — proNPY-like and NPY —).</note><note type="content">Fig. 6: Effect of the combined treatment with RA and TPA on NPY gene expression (Northern blot analysis). Cells were treated for 24 h with 10 μM RA, 20 nM TPA alone or in association. Total RNA (30 μg/lane) was analyzed by Northern blot hybridization (C=control; RA=all-trans retinoic acid). The stripped blots were rehybridized for GAPDH mRNA for normalization. One Northern blot, representative of three separate experiments, is shown.</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a">Retinoic acid negatively regulates neuropeptide Y expression in human neuroblastoma cells</title><author><persName><forename type="first">Paolo</forename><surname>Magni</surname></persName><email>paolo.magni@unimi.it</email><note type="correspondence"><p>Corresponding author. Tel.: +39-02-205213219; fax: +39-02-29404927</p></note><affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</affiliation></author><author><persName><forename type="first">Elena</forename><surname>Beretta</surname></persName><affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</affiliation></author><author><persName><forename type="first">Eugenia</forename><surname>Scaccianoce</surname></persName><affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</affiliation></author><author><persName><forename type="first">Marcella</forename><surname>Motta</surname></persName><affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</affiliation></author></analytic><monogr><title level="j">Neuropharmacology</title><title level="j" type="abbrev">NP</title><idno type="pISSN">0028-3908</idno><idno type="PII">S0028-3908(00)X0061-5</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1999"></date><biblScope unit="volume">39</biblScope><biblScope unit="issue">9</biblScope><biblScope unit="page" from="1628">1628</biblScope><biblScope unit="page" to="1636">1636</biblScope></imprint></monogr><idno type="istex">75363D06AF5372F57E111CEA14B2CFAE018A838A</idno><idno type="DOI">10.1016/S0028-3908(99)00231-2</idno><idno type="PII">S0028-3908(99)00231-2</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>2000</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Retinoids are involved in the regulation of development and differentiation in many tissues, including the nervous system, where they have been associated with some neurotransmitter systems. In the present study, we evaluated the effects of all-trans retinoic acid (RA) on the biosynthesis and secretion of neuropeptide Y (NPY), a widely expressed neuroregulatory peptide. The SH-SY5Y human neuroblastoma cell line has been used as the in vitro model system. Treatment with 10 μM RA induced a marked decrease in NPY gene expression after as little as 3–6 h of incubation and resulted in its almost complete suppression at 12–24 h and after a 6-day differentiating treatment. The NPY content in cell extracts and the NPY secreted and accumulated in the culture medium were also reduced by exposure to 10 μM RA at 12 and 24 h and at 6 days. Moreover, RA treatment for 6 days, but not for 24 h, resulted in a marked stimulation of proNPY processing to mature NPY. The presence of negative retinoic acid-response elements in the human NPY promoter (up to −1078 bp) was excluded by a computer search. When SH-SY5Y cells were treated simultaneously with 20 nM TPA and 10 μM RA for 24 h, the marked stimulatory effect of TPA alone was completely suppressed. These observations suggest that the expression of NPY in SH-SY5Y human neuroblastoma cells is negatively regulated by RA at the level of gene expression, probably by mechanisms involving the interaction of activated RARs with transcription factors (such as AP-1).</p></abstract><textClass><keywords scheme="keyword"><list><head>Keywords</head><item><term>Retinoic acid</term></item><item><term>Neuropeptide Y gene expression</term></item><item><term>Neuropeptide Y biosynthesis</term></item><item><term>Neuroblastoma</term></item><item><term>Cell differentiation</term></item><item><term>Phorbol ester</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1999-11-08">Registration</change><change when="1999">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><original>false</original><mimetype>text/plain</mimetype><extension>txt</extension><uri>https://api.istex.fr/document/75363D06AF5372F57E111CEA14B2CFAE018A838A/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: ce:floats; body; 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:entity SYSTEM="gr1" NDATA="IMAGE" name="gr1"></istex:entity><istex:entity SYSTEM="gr2" NDATA="IMAGE" name="gr2"></istex:entity><istex:entity SYSTEM="gr3" NDATA="IMAGE" name="gr3"></istex:entity><istex:entity SYSTEM="gr4" NDATA="IMAGE" name="gr4"></istex:entity><istex:entity SYSTEM="gr5" NDATA="IMAGE" name="gr5"></istex:entity><istex:entity SYSTEM="gr6" NDATA="IMAGE" name="gr6"></istex:entity></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla"><item-info><jid>NP</jid><aid>1280</aid><ce:pii>S0028-3908(99)00231-2</ce:pii><ce:doi>10.1016/S0028-3908(99)00231-2</ce:doi><ce:copyright type="full-transfer" year="2000">Elsevier Science Ltd</ce:copyright></item-info><head><ce:title>Retinoic acid negatively regulates neuropeptide Y expression in human neuroblastoma cells</ce:title><ce:author-group><ce:author><ce:given-name>Paolo</ce:given-name><ce:surname>Magni</ce:surname><ce:cross-ref refid="CORR1">*</ce:cross-ref><ce:e-address>paolo.magni@unimi.it</ce:e-address></ce:author><ce:author><ce:given-name>Elena</ce:given-name><ce:surname>Beretta</ce:surname></ce:author><ce:author><ce:given-name>Eugenia</ce:given-name><ce:surname>Scaccianoce</ce:surname></ce:author><ce:author><ce:given-name>Marcella</ce:given-name><ce:surname>Motta</ce:surname></ce:author><ce:affiliation><ce:textfn>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</ce:textfn></ce:affiliation><ce:correspondence id="CORR1"><ce:label>*</ce:label><ce:text>Corresponding author. Tel.: +39-02-205213219; fax: +39-02-29404927</ce:text></ce:correspondence></ce:author-group><ce:date-accepted day="8" month="11" year="1999"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>Retinoids are involved in the regulation of development and differentiation in many tissues, including the nervous system, where they have been associated with some neurotransmitter systems. In the present study, we evaluated the effects of all-<ce:italic>trans</ce:italic> retinoic acid (RA) on the biosynthesis and secretion of neuropeptide Y (NPY), a widely expressed neuroregulatory peptide. The SH-SY5Y human neuroblastoma cell line has been used as the in vitro model system. Treatment with 10 μM RA induced a marked decrease in NPY gene expression after as little as 3–6 h of incubation and resulted in its almost complete suppression at 12–24 h and after a 6-day differentiating treatment. The NPY content in cell extracts and the NPY secreted and accumulated in the culture medium were also reduced by exposure to 10 μM RA at 12 and 24 h and at 6 days. Moreover, RA treatment for 6 days, but not for 24 h, resulted in a marked stimulation of proNPY processing to mature NPY. The presence of negative retinoic acid-response elements in the human NPY promoter (up to −1078 bp) was excluded by a computer search. When SH-SY5Y cells were treated simultaneously with 20 nM TPA and 10 μM RA for 24 h, the marked stimulatory effect of TPA alone was completely suppressed. These observations suggest that the expression of NPY in SH-SY5Y human neuroblastoma cells is negatively regulated by RA at the level of gene expression, probably by mechanisms involving the interaction of activated RARs with transcription factors (such as AP-1).</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords class="keyword"><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>Retinoic acid</ce:text></ce:keyword><ce:keyword><ce:text>Neuropeptide Y gene expression</ce:text></ce:keyword><ce:keyword><ce:text>Neuropeptide Y biosynthesis</ce:text></ce:keyword><ce:keyword><ce:text>Neuroblastoma</ce:text></ce:keyword><ce:keyword><ce:text>Cell differentiation</ce:text></ce:keyword><ce:keyword><ce:text>Phorbol ester</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Retinoic acid negatively regulates neuropeptide Y expression in human neuroblastoma cells</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Retinoic acid negatively regulates neuropeptide Y expression in human neuroblastoma cells</title></titleInfo><name type="personal"><namePart type="given">Paolo</namePart><namePart type="family">Magni</namePart><affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</affiliation><affiliation>E-mail: paolo.magni@unimi.it</affiliation><description>Corresponding author. Tel.: +39-02-205213219; fax: +39-02-29404927</description><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Elena</namePart><namePart type="family">Beretta</namePart><affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Eugenia</namePart><namePart type="family">Scaccianoce</namePart><affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Marcella</namePart><namePart type="family">Motta</namePart><affiliation>Center for Endocrinological Oncology, Institute of Endocrinology, University of Milan, Milan, Italy</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">1999</dateIssued><dateValid encoding="w3cdtf">1999-11-08</dateValid><copyrightDate encoding="w3cdtf">2000</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">Retinoids are involved in the regulation of development and differentiation in many tissues, including the nervous system, where they have been associated with some neurotransmitter systems. In the present study, we evaluated the effects of all-trans retinoic acid (RA) on the biosynthesis and secretion of neuropeptide Y (NPY), a widely expressed neuroregulatory peptide. The SH-SY5Y human neuroblastoma cell line has been used as the in vitro model system. Treatment with 10 μM RA induced a marked decrease in NPY gene expression after as little as 3–6 h of incubation and resulted in its almost complete suppression at 12–24 h and after a 6-day differentiating treatment. The NPY content in cell extracts and the NPY secreted and accumulated in the culture medium were also reduced by exposure to 10 μM RA at 12 and 24 h and at 6 days. Moreover, RA treatment for 6 days, but not for 24 h, resulted in a marked stimulation of proNPY processing to mature NPY. The presence of negative retinoic acid-response elements in the human NPY promoter (up to −1078 bp) was excluded by a computer search. When SH-SY5Y cells were treated simultaneously with 20 nM TPA and 10 μM RA for 24 h, the marked stimulatory effect of TPA alone was completely suppressed. These observations suggest that the expression of NPY in SH-SY5Y human neuroblastoma cells is negatively regulated by RA at the level of gene expression, probably by mechanisms involving the interaction of activated RARs with transcription factors (such as AP-1).</abstract><note type="content">Fig. 1: Effect of treatment with RA on NPY gene expression (Northern blot analysis). Cells were treated for 3, 6, 12, 24 h and six days with 10 μM RA. Total RNA (30 μg/lane) was analyzed by Northern blot hybridization (C=control; RA=all-trans retinoic acid). The stripped blots were rehybridized for GAPDH mRNA for normalization. One Northern blot, representative of three separate experiments, is shown.</note><note type="content">Fig. 2: Effect of the combined treatment with RA and TPA on NPY gene expression (Northern blot analysis). Cells were treated for 24 h with RA at different concentrations, in the presence of 20 nM TPA, or in its absence. Total RNA (30 μg/lane) was analyzed by Northern blot hybridization (C=control; RA=all-trans retinoic acid). The stripped blots were rehybridized for GAPDH mRNA for normalization. One Northern blot, representative of three separate experiments, is shown.</note><note type="content">Fig. 3: Morphological changes of SH-SY5Y human neuroblastoma cells after differentiation with RA. Cells were treated for six days with 10 μM RA (all-trans retinoic acid) or solvent (C=control). The effect of treatment with 20 nM TPA for six days is shown for comparison.</note><note type="content">Fig. 4: Effect of treatment with RA on NPY synthesis/secretion. Cells were treated for 3, 6, 12, 24 h and six days with 10 μM RA and the NPY-IR content in cell extracts and in the medium was quantified by RIA. Panel A: effect of RA treatment on NPY-IR content in cell extracts. Panel B: effect of RA treatment on NPY-IR content in the medium. One experiment representative of three or four separate experiments is shown. Data are expressed as mean±S.E.M. (percent of controls=100). C=control; RA=all-trans retinoic acid; n=6; *P<0.05 vs C; **P<0.01 vs C; ***P<0.001 vs C (ANOVA).</note><note type="content">Fig. 5: Effect of 24-h and 6-day treatments with 10 μM RA on NPY precursor processing (cell extracts and conditioned medium). One experiment representative of 3–5 separate experiments is shown. Cell extracts (panel A) and conditioned media (panel B) were size-fractionated by gel filtration chromatography and the NPY-IR in each fraction was quantified by RIA. Dotted line: C (control); solid line: RA (all-trans retinoic acid). Please note that, for the purpose of clarity, the elution profiles of samples from cells treated for 24 h and for six days are separately superimposed to the same control elution profile (dotted line). In the upper left and in the lower right profiles is indicated the elution position of synthetic human NPY (“NPY std”), used as the reference standard, and the immunoreactive peak corresponding to the proNPY-like substance (“proNPY-like”). The table under each panel summarizes the data of gel filtration chromatography obtained in all the experiments performed and includes the representative chromatograms shown above them. The data (mean±S.E.M.) relative to the proNPY-like and to the NPY peaks are expressed as % of total NPY-IR (total NPY-IR=sum of the areas under the curve relative to the two chromatographic peaks — proNPY-like and NPY —).</note><note type="content">Fig. 6: Effect of the combined treatment with RA and TPA on NPY gene expression (Northern blot analysis). Cells were treated for 24 h with 10 μM RA, 20 nM TPA alone or in association. Total RNA (30 μg/lane) was analyzed by Northern blot hybridization (C=control; RA=all-trans retinoic acid). The stripped blots were rehybridized for GAPDH mRNA for normalization. One Northern blot, representative of three separate experiments, is shown.</note><subject><genre>Keywords</genre><topic>Retinoic acid</topic><topic>Neuropeptide Y gene expression</topic><topic>Neuropeptide Y biosynthesis</topic><topic>Neuroblastoma</topic><topic>Cell differentiation</topic><topic>Phorbol ester</topic></subject><relatedItem type="host"><titleInfo><title>Neuropharmacology</title></titleInfo><titleInfo type="abbreviated"><title>NP</title></titleInfo><genre type="Journal">journal</genre><originInfo><dateIssued encoding="w3cdtf">200008</dateIssued></originInfo><identifier type="ISSN">0028-3908</identifier><identifier type="PII">S0028-3908(00)X0061-5</identifier><part><date>200008</date><detail type="volume"><number>39</number><caption>vol.</caption></detail><detail type="issue"><number>9</number><caption>no.</caption></detail><extent unit="issue pages"><start>1483</start><end>1680</end></extent><extent unit="pages"><start>1628</start><end>1636</end></extent></part></relatedItem><identifier type="istex">75363D06AF5372F57E111CEA14B2CFAE018A838A</identifier><identifier type="DOI">10.1016/S0028-3908(99)00231-2</identifier><identifier type="PII">S0028-3908(99)00231-2</identifier><accessCondition type="use and reproduction" contentType="">© 2000Elsevier Science Ltd</accessCondition><recordInfo><recordContentSource>ELSEVIER</recordContentSource><recordOrigin>Elsevier Science Ltd, ©2000</recordOrigin></recordInfo></mods></metadata><enrichments><istex:catWosTEI uri="https://api.istex.fr/document/75363D06AF5372F57E111CEA14B2CFAE018A838A/enrichments/catWos"><teiHeader><profileDesc><textClass><classCode scheme="WOS">PHARMACOLOGY & PHARMACY</classCode><classCode scheme="WOS">NEUROSCIENCES</classCode></textClass></profileDesc></teiHeader></istex:catWosTEI></enrichments><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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