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Molecular characterization of commonly used cell lines for bone tumor research: A trans‐European EuroBoNet effort

Identifieur interne : 001800 ( Istex/Curation ); précédent : 001799; suivant : 001801

Molecular characterization of commonly used cell lines for bone tumor research: A trans‐European EuroBoNet effort

Auteurs : Laura Ottaviano [Allemagne] ; Karl-Ludwig Schaefer [Allemagne] ; Melanie Gajewski [Allemagne] ; Wolfgang Huckenbeck [Allemagne] ; Stefan Baldus [Allemagne] ; Uwe Rogel [Allemagne] ; Carlos Mackintosh [Espagne] ; Enrique De Alava [Espagne] ; Ola Myklebost [Norvège] ; Stine H. Kresse [Norvège] ; Leonardo A. Meza-Zepeda [Norvège] ; Massimo Serra [Italie] ; Anne-Marie Cleton-Jansen [Pays-Bas] ; Pancras C. W. Hogendoorn [Pays-Bas] ; Horst Buerger [Allemagne] ; Thomas Aigner [Allemagne] ; Helmut E. Gabbert [Allemagne] ; Christopher Poremba [Allemagne]

Source :

RBID : ISTEX:E8374F395D7E218EE22C21CB4F7C9DACF7D1C4D9

Abstract

Usage of cancer cell lines has repeatedly generated conflicting results provoked by differences among subclones or contamination with mycoplasm or other immortal mammalian cells. To overcome these limitations, we decided within the EuroBoNeT consortium to characterize a common set of cell lines including osteosarcomas (OS), Ewing sarcomas (ES), and chondrosarcomas (CS). DNA fingerprinting was used to guarantee the identity of all of the cell lines and to distinguish subclones of osteosarcoma cell line HOS. Screening for homozygous loss of 38 tumor suppressor genes by MLPA revealed deletion of CDKN2A as the most common event (15/36), strictly associated with absence of the CDKN2A (p16) protein. Ten cell lines showed missense mutations of the TP53 gene while another set of nine cell lines showed mutations resulting in truncation of the TP53 protein. Cells harboring missense mutations expressed high levels of nuclear TP53, while cell lines with nonsense mutations showed weak/absent staining for TP53. TP53wt cell lines usually expressed the protein in 2–10% of the cells. However, seven TP53wt osteosarcomas were negative for both mRNA and protein expression. Our analyses shed light on the correlation between immunohistochemical and genetic data for CDKN2A and TP53, and confirm the importance of these signaling pathways. The characterization of a substantial number of cell lines represents an important step to supply research groups with proven models for further advanced studies on tumor biology and may help to make results from different laboratories more comparable. © 2009 Wiley‐Liss, Inc.

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DOI: 10.1002/gcc.20717

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ISTEX:E8374F395D7E218EE22C21CB4F7C9DACF7D1C4D9

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<name sortKey="Myklebost, Ola" sort="Myklebost, Ola" uniqKey="Myklebost O" first="Ola" last="Myklebost">Ola Myklebost</name>
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<name sortKey="Serra, Massimo" sort="Serra, Massimo" uniqKey="Serra M" first="Massimo" last="Serra">Massimo Serra</name>
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<name sortKey="Cleton Ansen, Anne Arie" sort="Cleton Ansen, Anne Arie" uniqKey="Cleton Ansen A" first="Anne-Marie" last="Cleton-Jansen">Anne-Marie Cleton-Jansen</name>
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<mods:affiliation>Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands</mods:affiliation>
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<name sortKey="Poremba, Christopher" sort="Poremba, Christopher" uniqKey="Poremba C" first="Christopher" last="Poremba">Christopher Poremba</name>
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<name sortKey="Gajewski, Melanie" sort="Gajewski, Melanie" uniqKey="Gajewski M" first="Melanie" last="Gajewski">Melanie Gajewski</name>
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<name sortKey="Huckenbeck, Wolfgang" sort="Huckenbeck, Wolfgang" uniqKey="Huckenbeck W" first="Wolfgang" last="Huckenbeck">Wolfgang Huckenbeck</name>
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<name sortKey="Baldus, Stefan" sort="Baldus, Stefan" uniqKey="Baldus S" first="Stefan" last="Baldus">Stefan Baldus</name>
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<name sortKey="Rogel, Uwe" sort="Rogel, Uwe" uniqKey="Rogel U" first="Uwe" last="Rogel">Uwe Rogel</name>
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<mods:affiliation>Institute of Pathology, University Medical Center Duesseldorf, Duesseldorf, Germany</mods:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute of Pathology, University Medical Center Duesseldorf, Duesseldorf</wicri:regionArea>
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<name sortKey="Mackintosh, Carlos" sort="Mackintosh, Carlos" uniqKey="Mackintosh C" first="Carlos" last="Mackintosh">Carlos Mackintosh</name>
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<mods:affiliation>Centro de Investigación del Cáncer‐IBMCC, Universidad de Salamanca‐CSIC, Salamanca, Spain</mods:affiliation>
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<name sortKey="De Alava, Enrique" sort="De Alava, Enrique" uniqKey="De Alava E" first="Enrique" last="De Alava">Enrique De Alava</name>
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<mods:affiliation>Centro de Investigación del Cáncer‐IBMCC, Universidad de Salamanca‐CSIC, Salamanca, Spain</mods:affiliation>
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<name sortKey="Myklebost, Ola" sort="Myklebost, Ola" uniqKey="Myklebost O" first="Ola" last="Myklebost">Ola Myklebost</name>
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<name sortKey="Kresse, Stine H" sort="Kresse, Stine H" uniqKey="Kresse S" first="Stine H." last="Kresse">Stine H. Kresse</name>
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<mods:affiliation>Department of Tumor Biology, The Norwegian Radium Hospital, Rikshospitalet University Hospital, Oslo, Norway</mods:affiliation>
<country xml:lang="fr">Norvège</country>
<wicri:regionArea>Department of Tumor Biology, The Norwegian Radium Hospital, Rikshospitalet University Hospital, Oslo</wicri:regionArea>
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<name sortKey="Meza Epeda, Leonardo A" sort="Meza Epeda, Leonardo A" uniqKey="Meza Epeda L" first="Leonardo A." last="Meza-Zepeda">Leonardo A. Meza-Zepeda</name>
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<mods:affiliation>Department of Tumor Biology, The Norwegian Radium Hospital, Rikshospitalet University Hospital, Oslo, Norway</mods:affiliation>
<country xml:lang="fr">Norvège</country>
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<name sortKey="Serra, Massimo" sort="Serra, Massimo" uniqKey="Serra M" first="Massimo" last="Serra">Massimo Serra</name>
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<mods:affiliation>Laboratorio di Ricerca Oncologica, Istituti Ortopedici Rizzoli, Bologna, Italy</mods:affiliation>
<country xml:lang="fr">Italie</country>
<wicri:regionArea>Laboratorio di Ricerca Oncologica, Istituti Ortopedici Rizzoli, Bologna</wicri:regionArea>
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<name sortKey="Cleton Ansen, Anne Arie" sort="Cleton Ansen, Anne Arie" uniqKey="Cleton Ansen A" first="Anne-Marie" last="Cleton-Jansen">Anne-Marie Cleton-Jansen</name>
<affiliation wicri:level="1">
<mods:affiliation>Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands</mods:affiliation>
<country xml:lang="fr">Pays-Bas</country>
<wicri:regionArea>Department of Pathology, Leiden University Medical Center, Leiden</wicri:regionArea>
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<name sortKey="Hogendoorn, Pancras C W" sort="Hogendoorn, Pancras C W" uniqKey="Hogendoorn P" first="Pancras C. W." last="Hogendoorn">Pancras C. W. Hogendoorn</name>
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<mods:affiliation>Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands</mods:affiliation>
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<wicri:regionArea>Department of Pathology, Leiden University Medical Center, Leiden</wicri:regionArea>
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<name sortKey="Buerger, Horst" sort="Buerger, Horst" uniqKey="Buerger H" first="Horst" last="Buerger">Horst Buerger</name>
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<mods:affiliation>Institute of Pathology, University Medical Center Muenster, Muenster, Germany</mods:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute of Pathology, University Medical Center Muenster, Muenster</wicri:regionArea>
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<name sortKey="Aigner, Thomas" sort="Aigner, Thomas" uniqKey="Aigner T" first="Thomas" last="Aigner">Thomas Aigner</name>
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<mods:affiliation>Institute of Pathology, University of Leipzig, Leipzig, Germany</mods:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute of Pathology, University of Leipzig, Leipzig</wicri:regionArea>
</affiliation>
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<name sortKey="Gabbert, Helmut E" sort="Gabbert, Helmut E" uniqKey="Gabbert H" first="Helmut E." last="Gabbert">Helmut E. Gabbert</name>
<affiliation wicri:level="1">
<mods:affiliation>Institute of Pathology, University Medical Center Duesseldorf, Duesseldorf, Germany</mods:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute of Pathology, University Medical Center Duesseldorf, Duesseldorf</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Poremba, Christopher" sort="Poremba, Christopher" uniqKey="Poremba C" first="Christopher" last="Poremba">Christopher Poremba</name>
<affiliation wicri:level="1">
<mods:affiliation>Institute of Pathology, University Medical Center Duesseldorf, Duesseldorf, Germany</mods:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute of Pathology, University Medical Center Duesseldorf, Duesseldorf</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1">
<mods:affiliation>Center of Histopathology, Cytology and Molecular Diagnostics (CHCMD), Research Park Trier, Trier, Germany</mods:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Center of Histopathology, Cytology and Molecular Diagnostics (CHCMD), Research Park Trier, Trier</wicri:regionArea>
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<monogr></monogr>
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<title level="j">Genes, Chromosomes and Cancer</title>
<title level="j" type="abbrev">Genes Chromosom. Cancer</title>
<idno type="ISSN">1045-2257</idno>
<idno type="eISSN">1098-2264</idno>
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<publisher>Wiley Subscription Services, Inc., A Wiley Company</publisher>
<pubPlace>Hoboken</pubPlace>
<date type="published" when="2010-01">2010-01</date>
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<idno type="DOI">10.1002/gcc.20717</idno>
<idno type="ArticleID">GCC20717</idno>
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<idno type="ISSN">1045-2257</idno>
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<div type="abstract" xml:lang="en">Usage of cancer cell lines has repeatedly generated conflicting results provoked by differences among subclones or contamination with mycoplasm or other immortal mammalian cells. To overcome these limitations, we decided within the EuroBoNeT consortium to characterize a common set of cell lines including osteosarcomas (OS), Ewing sarcomas (ES), and chondrosarcomas (CS). DNA fingerprinting was used to guarantee the identity of all of the cell lines and to distinguish subclones of osteosarcoma cell line HOS. Screening for homozygous loss of 38 tumor suppressor genes by MLPA revealed deletion of CDKN2A as the most common event (15/36), strictly associated with absence of the CDKN2A (p16) protein. Ten cell lines showed missense mutations of the TP53 gene while another set of nine cell lines showed mutations resulting in truncation of the TP53 protein. Cells harboring missense mutations expressed high levels of nuclear TP53, while cell lines with nonsense mutations showed weak/absent staining for TP53. TP53wt cell lines usually expressed the protein in 2–10% of the cells. However, seven TP53wt osteosarcomas were negative for both mRNA and protein expression. Our analyses shed light on the correlation between immunohistochemical and genetic data for CDKN2A and TP53, and confirm the importance of these signaling pathways. The characterization of a substantial number of cell lines represents an important step to supply research groups with proven models for further advanced studies on tumor biology and may help to make results from different laboratories more comparable. © 2009 Wiley‐Liss, Inc.</div>
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