Viral load, gene expression and mapping of viral integration sites in HPV16-associated HNSCC cell lines
Identifieur interne : 000865 ( Main/Exploration ); précédent : 000864; suivant : 000866Viral load, gene expression and mapping of viral integration sites in HPV16-associated HNSCC cell lines
Auteurs : Nadine C. Olthof [Pays-Bas] ; Christian U. Huebbers [Allemagne] ; Jutta Kolligs [Allemagne] ; Mieke Henfling [Pays-Bas] ; Frans C. S. Ramaekers [Pays-Bas] ; Iris Cornet [Pays-Bas] ; Josefa A. Van Lent-Albrechts [Pays-Bas] ; Sander P. A. Stegmann [Pays-Bas] ; Steffi Silling [Allemagne] ; Ulrike Wieland [Allemagne] ; Thomas E. Carey [États-Unis] ; Heather M. Walline [États-Unis] ; Susanne M. Gollin [États-Unis] ; Thomas K. Hoffmann [Allemagne] ; Johan De Winter [Pays-Bas] ; Bernd Kremer [Pays-Bas] ; Jens-Peter Klussmann [Allemagne] ; Ernst-Jan M. Speel [Pays-Bas]Source :
- International journal of cancer [ 0020-7136 ] ; 2014.
Abstract
HPV-related HNSCC generally have a better prognosis than HPV-negative HNSCC. However, a subgroup of HPV-positive tumors with poor prognosis has been recognized, particularly related to smoking, EGFR overexpression and chromosomal instability. Viral integration into the host genome might contribute to carcinogenesis, as is shown for cervical carcinomas. Therefore, all HPV16-positive HNSCC cell lines currently available have been carefully analysed for viral and host genome parameters.
The viral integration status, viral load, viral gene expression and the presence of aneusomies was evaluated in the cell lines UD-SCC-2, UM-SCC-047, UM-SCC-104, UPCI:SCC090, UPCI:SCC152, UPCI:SCC154 and 93VU147T. HPV integration was examined using FISH, APOT-PCR and DIPS-PCR. Viral load and the expression of the viral genes E2, E6 and E7 were determined via quantitative PCR.
All cell lines showed integration-specific staining patterns and signals indicating transcriptional activity using FISH. APOT- and DIPS-PCR identified integration-derived fusion products in six cell lines, and only episomal products for UM-SCC-104. Despite the observed differences in viral load and the number of viral integration sites, this did not relate to the identified viral oncogene expression. Furthermore, cell lines exhibited EGFR expression, and aneusomy (except UPCI:SCC154).
In conclusion, all HPV16-positive HNSCC cell lines showed integrated and/or episomal viral DNA that is transcriptionally active, although viral oncogene expression was independent of viral copy number and the number of viral integration sites. Because these cell lines also contain EGFR expression and aneusomy, which are parameters of poor prognosis, they should be considered suitable model systems for the development of new antiviral therapies.
Url:
DOI: 10.1002/ijc.29112
PubMed: 25082736
PubMed Central: 5370555
Affiliations:
- Allemagne, Pays-Bas, États-Unis
- Bade-Wurtemberg, District de Tübingen, Hollande-Septentrionale, Pennsylvanie
- Amsterdam, Ulm
Links toward previous steps (curation, corpus...)
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- to stream Ncbi, to step Merge: 002D21
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Le document en format XML
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<affiliation wicri:level="1"><nlm:aff id="A6">Institute of Virology, National Reference Centre for Papilloma- and Polyomaviruses, University Hospital of Cologne, Cologne, Germany</nlm:aff>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute of Virology, National Reference Centre for Papilloma- and Polyomaviruses, University Hospital of Cologne, Cologne</wicri:regionArea>
<wicri:noRegion>Cologne</wicri:noRegion>
<wicri:noRegion>Cologne</wicri:noRegion>
<wicri:noRegion>Cologne</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Carey, Thomas E" sort="Carey, Thomas E" uniqKey="Carey T" first="Thomas E." last="Carey">Thomas E. Carey</name>
<affiliation wicri:level="1"><nlm:aff id="A7">Department of Otolaryngology, University of Michigan, Ann Arbor, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Otolaryngology, University of Michigan, Ann Arbor</wicri:regionArea>
<wicri:noRegion>Ann Arbor</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Walline, Heather M" sort="Walline, Heather M" uniqKey="Walline H" first="Heather M." last="Walline">Heather M. Walline</name>
<affiliation wicri:level="1"><nlm:aff id="A7">Department of Otolaryngology, University of Michigan, Ann Arbor, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Otolaryngology, University of Michigan, Ann Arbor</wicri:regionArea>
<wicri:noRegion>Ann Arbor</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Gollin, Susanne M" sort="Gollin, Susanne M" uniqKey="Gollin S" first="Susanne M." last="Gollin">Susanne M. Gollin</name>
<affiliation wicri:level="2"><nlm:aff id="A8">Department of Human Genetics, University of Pittsburgh Graduate School of Public Health and the University of Pittsburgh Cancer Institute, PA, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Human Genetics, University of Pittsburgh Graduate School of Public Health and the University of Pittsburgh Cancer Institute, PA</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Hoffmann, Thomas K" sort="Hoffmann, Thomas K" uniqKey="Hoffmann T" first="Thomas K." last="Hoffmann">Thomas K. Hoffmann</name>
<affiliation wicri:level="3"><nlm:aff id="A9">Department of Otorhinolaryngology and Head and Neck Surgery, Ulm, Germany</nlm:aff>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Department of Otorhinolaryngology and Head and Neck Surgery, Ulm</wicri:regionArea>
<placeName><region type="land" nuts="1">Bade-Wurtemberg</region>
<region type="district" nuts="2">District de Tübingen</region>
<settlement type="city">Ulm</settlement>
</placeName>
</affiliation>
</author>
<author><name sortKey="De Winter, Johan" sort="De Winter, Johan" uniqKey="De Winter J" first="Johan" last="De Winter">Johan De Winter</name>
<affiliation wicri:level="3"><nlm:aff id="A10">Department of Clinical Genetics, VU University Medical Center, Amsterdam, the Netherlands</nlm:aff>
<country xml:lang="fr" wicri:curation="lc">Pays-Bas</country>
<wicri:regionArea>Department of Clinical Genetics, VU University Medical Center, Amsterdam</wicri:regionArea>
<placeName><settlement type="city">Amsterdam</settlement>
<region nuts="2" type="province">Hollande-Septentrionale</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Kremer, Bernd" sort="Kremer, Bernd" uniqKey="Kremer B" first="Bernd" last="Kremer">Bernd Kremer</name>
<affiliation wicri:level="1"><nlm:aff id="A1">Department of Otorhinolaryngology and Head and Neck Surgery, Maastricht University Medical Centre, Maastricht, the Netherlands</nlm:aff>
<country xml:lang="fr" wicri:curation="lc">Pays-Bas</country>
<wicri:regionArea>Department of Otorhinolaryngology and Head and Neck Surgery, Maastricht University Medical Centre, Maastricht</wicri:regionArea>
<wicri:noRegion>Maastricht</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Klussmann, Jens Peter" sort="Klussmann, Jens Peter" uniqKey="Klussmann J" first="Jens-Peter" last="Klussmann">Jens-Peter Klussmann</name>
<affiliation wicri:level="1"><nlm:aff id="A11">Department of Otorhinolaryngology, Head and Neck Surgery, School of Medicine, University Hospital of Giessen, Giessen, Germany</nlm:aff>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Department of Otorhinolaryngology, Head and Neck Surgery, School of Medicine, University Hospital of Giessen, Giessen</wicri:regionArea>
<wicri:noRegion>Giessen</wicri:noRegion>
<wicri:noRegion>Giessen</wicri:noRegion>
<wicri:noRegion>Giessen</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Speel, Ernst Jan M" sort="Speel, Ernst Jan M" uniqKey="Speel E" first="Ernst-Jan M." last="Speel">Ernst-Jan M. Speel</name>
<affiliation wicri:level="1"><nlm:aff id="A2">Department of Molecular Cell Biology, Maastricht University Medical Centre, Maastricht, the Netherlands</nlm:aff>
<country xml:lang="fr" wicri:curation="lc">Pays-Bas</country>
<wicri:regionArea>Department of Molecular Cell Biology, Maastricht University Medical Centre, Maastricht</wicri:regionArea>
<wicri:noRegion>Maastricht</wicri:noRegion>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="A4">Department of Pathology, Maastricht University Medical Centre, Maastricht, the Netherlands</nlm:aff>
<country xml:lang="fr" wicri:curation="lc">Pays-Bas</country>
<wicri:regionArea>Department of Pathology, Maastricht University Medical Centre, Maastricht</wicri:regionArea>
<wicri:noRegion>Maastricht</wicri:noRegion>
</affiliation>
</author>
</analytic>
<series><title level="j">International journal of cancer</title>
<idno type="ISSN">0020-7136</idno>
<idno type="eISSN">1097-0215</idno>
<imprint><date when="2014">2014</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><p id="P1">HPV-related HNSCC generally have a better prognosis than HPV-negative HNSCC. However, a subgroup of HPV-positive tumors with poor prognosis has been recognized, particularly related to smoking, EGFR overexpression and chromosomal instability. Viral integration into the host genome might contribute to carcinogenesis, as is shown for cervical carcinomas. Therefore, all HPV16-positive HNSCC cell lines currently available have been carefully analysed for viral and host genome parameters.</p>
<p id="P2">The viral integration status, viral load, viral gene expression and the presence of aneusomies was evaluated in the cell lines UD-SCC-2, UM-SCC-047, UM-SCC-104, UPCI:SCC090, UPCI:SCC152, UPCI:SCC154 and 93VU147T. HPV integration was examined using FISH, APOT-PCR and DIPS-PCR. Viral load and the expression of the viral genes E2, E6 and E7 were determined via quantitative PCR.</p>
<p id="P3">All cell lines showed integration-specific staining patterns and signals indicating transcriptional activity using FISH. APOT- and DIPS-PCR identified integration-derived fusion products in six cell lines, and only episomal products for UM-SCC-104. Despite the observed differences in viral load and the number of viral integration sites, this did not relate to the identified viral oncogene expression. Furthermore, cell lines exhibited EGFR expression, and aneusomy (except UPCI:SCC154).</p>
<p id="P4">In conclusion, all HPV16-positive HNSCC cell lines showed integrated and/or episomal viral DNA that is transcriptionally active, although viral oncogene expression was independent of viral copy number and the number of viral integration sites. Because these cell lines also contain EGFR expression and aneusomy, which are parameters of poor prognosis, they should be considered suitable model systems for the development of new antiviral therapies.</p>
</div>
</front>
</TEI>
<affiliations><list><country><li>Allemagne</li>
<li>Pays-Bas</li>
<li>États-Unis</li>
</country>
<region><li>Bade-Wurtemberg</li>
<li>District de Tübingen</li>
<li>Hollande-Septentrionale</li>
<li>Pennsylvanie</li>
</region>
<settlement><li>Amsterdam</li>
<li>Ulm</li>
</settlement>
</list>
<tree><country name="Pays-Bas"><noRegion><name sortKey="Olthof, Nadine C" sort="Olthof, Nadine C" uniqKey="Olthof N" first="Nadine C." last="Olthof">Nadine C. Olthof</name>
</noRegion>
<name sortKey="Cornet, Iris" sort="Cornet, Iris" uniqKey="Cornet I" first="Iris" last="Cornet">Iris Cornet</name>
<name sortKey="Cornet, Iris" sort="Cornet, Iris" uniqKey="Cornet I" first="Iris" last="Cornet">Iris Cornet</name>
<name sortKey="De Winter, Johan" sort="De Winter, Johan" uniqKey="De Winter J" first="Johan" last="De Winter">Johan De Winter</name>
<name sortKey="Henfling, Mieke" sort="Henfling, Mieke" uniqKey="Henfling M" first="Mieke" last="Henfling">Mieke Henfling</name>
<name sortKey="Kremer, Bernd" sort="Kremer, Bernd" uniqKey="Kremer B" first="Bernd" last="Kremer">Bernd Kremer</name>
<name sortKey="Olthof, Nadine C" sort="Olthof, Nadine C" uniqKey="Olthof N" first="Nadine C." last="Olthof">Nadine C. Olthof</name>
<name sortKey="Ramaekers, Frans C S" sort="Ramaekers, Frans C S" uniqKey="Ramaekers F" first="Frans C. S." last="Ramaekers">Frans C. S. Ramaekers</name>
<name sortKey="Speel, Ernst Jan M" sort="Speel, Ernst Jan M" uniqKey="Speel E" first="Ernst-Jan M." last="Speel">Ernst-Jan M. Speel</name>
<name sortKey="Speel, Ernst Jan M" sort="Speel, Ernst Jan M" uniqKey="Speel E" first="Ernst-Jan M." last="Speel">Ernst-Jan M. Speel</name>
<name sortKey="Stegmann, Sander P A" sort="Stegmann, Sander P A" uniqKey="Stegmann S" first="Sander P. A." last="Stegmann">Sander P. A. Stegmann</name>
<name sortKey="Van Lent Albrechts, Josefa A" sort="Van Lent Albrechts, Josefa A" uniqKey="Van Lent Albrechts J" first="Josefa A." last="Van Lent-Albrechts">Josefa A. Van Lent-Albrechts</name>
</country>
<country name="Allemagne"><noRegion><name sortKey="Huebbers, Christian U" sort="Huebbers, Christian U" uniqKey="Huebbers C" first="Christian U." last="Huebbers">Christian U. Huebbers</name>
</noRegion>
<name sortKey="Hoffmann, Thomas K" sort="Hoffmann, Thomas K" uniqKey="Hoffmann T" first="Thomas K." last="Hoffmann">Thomas K. Hoffmann</name>
<name sortKey="Klussmann, Jens Peter" sort="Klussmann, Jens Peter" uniqKey="Klussmann J" first="Jens-Peter" last="Klussmann">Jens-Peter Klussmann</name>
<name sortKey="Kolligs, Jutta" sort="Kolligs, Jutta" uniqKey="Kolligs J" first="Jutta" last="Kolligs">Jutta Kolligs</name>
<name sortKey="Silling, Steffi" sort="Silling, Steffi" uniqKey="Silling S" first="Steffi" last="Silling">Steffi Silling</name>
<name sortKey="Wieland, Ulrike" sort="Wieland, Ulrike" uniqKey="Wieland U" first="Ulrike" last="Wieland">Ulrike Wieland</name>
</country>
<country name="États-Unis"><noRegion><name sortKey="Carey, Thomas E" sort="Carey, Thomas E" uniqKey="Carey T" first="Thomas E." last="Carey">Thomas E. Carey</name>
</noRegion>
<name sortKey="Gollin, Susanne M" sort="Gollin, Susanne M" uniqKey="Gollin S" first="Susanne M." last="Gollin">Susanne M. Gollin</name>
<name sortKey="Walline, Heather M" sort="Walline, Heather M" uniqKey="Walline H" first="Heather M." last="Walline">Heather M. Walline</name>
</country>
</tree>
</affiliations>
</record>
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