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Glioblastoma-derived spheroid cultures as an experimental model for analysis of EGFR anomalies
Glioblastoma cell cultures in vitro are frequently used for investigations on the biology of tumors or new therapeutic approaches. Recent reports have emphasized the importance of cell culture type for maintenance of tumor original features. Nevertheless, the ability of GBM cells to preserve EGFR ov...
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| Published in: | Journal of neuro-oncology 2011-05, Vol.102 (3), p.395-407 |
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| container_title | Journal of neuro-oncology |
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| creator | Witusik-Perkowska, Monika Rieske, Piotr Hułas-Bigoszewska, Krystyna Zakrzewska, Magdalena Stawski, Robert Kulczycka-Wojdala, Dominika Bieńkowski, Michał Stoczyńska-Fidelus, Ewelina Grešner, Sylwia M. Piaskowski, Sylwester Jaskólski, Dariusz J. Papierz, Wielisław Zakrzewski, Krzysztof Kolasa, Maciej Ironside, James W. Liberski, Paweł P. |
| description | Glioblastoma cell cultures in vitro are frequently used for investigations on the biology of tumors or new therapeutic approaches. Recent reports have emphasized the importance of cell culture type for maintenance of tumor original features. Nevertheless, the ability of GBM cells to preserve EGFR overdosage in vitro remains controversial. Our experimental approach was based on quantitative analysis of
EGFR
gene dosage in vitro both at DNA and mRNA level. Real-time PCR data were verified with a FISH method allowing for a distinction between
EGFR
amplification and polysomy 7. We demonstrated that
EGFR
amplification accompanied by
EGFRwt
overexpression was maintained in spheroids, but these phenomena were gradually lost in adherent culture. We noticed a rapid decrease of
EGFR
overdosage already at the initial stage of cell culture establishment. In contrast to
EGFR
amplification, the maintenance of polysomy 7 resulted in
EGFR
locus gain and stabilization even in long-term adherent culture in serum presence. Surprisingly, the
EGFRwt
expression pattern did not reflect the latter phenomenon and we observed no overexpression of the tested gene. Moreover, quantitative analysis demonstrated that expression of the truncated variant of receptor—
EGFRvIII
was preserved in GBM-derived spheroids at a level comparable to the initial tumor tissue. Our findings are especially important in the light of research using glioblastoma culture as the experimental model for testing novel EGFR-targeted therapeutics in vitro, with special emphasis on the most common mutated form of receptor—EGFRvIII. |
| doi_str_mv | 10.1007/s11060-010-0352-0 |
| format | article |
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EGFR
gene dosage in vitro both at DNA and mRNA level. Real-time PCR data were verified with a FISH method allowing for a distinction between
EGFR
amplification and polysomy 7. We demonstrated that
EGFR
amplification accompanied by
EGFRwt
overexpression was maintained in spheroids, but these phenomena were gradually lost in adherent culture. We noticed a rapid decrease of
EGFR
overdosage already at the initial stage of cell culture establishment. In contrast to
EGFR
amplification, the maintenance of polysomy 7 resulted in
EGFR
locus gain and stabilization even in long-term adherent culture in serum presence. Surprisingly, the
EGFRwt
expression pattern did not reflect the latter phenomenon and we observed no overexpression of the tested gene. Moreover, quantitative analysis demonstrated that expression of the truncated variant of receptor—
EGFRvIII
was preserved in GBM-derived spheroids at a level comparable to the initial tumor tissue. Our findings are especially important in the light of research using glioblastoma culture as the experimental model for testing novel EGFR-targeted therapeutics in vitro, with special emphasis on the most common mutated form of receptor—EGFRvIII.</description><identifier>ISSN: 0167-594X</identifier><identifier>EISSN: 1573-7373</identifier><identifier>DOI: 10.1007/s11060-010-0352-0</identifier><identifier>PMID: 20803305</identifier><language>eng</language><publisher>Boston: Springer US</publisher><subject>Animals ; Brain Neoplasms - pathology ; Bromodeoxyuridine - metabolism ; Cell Adhesion - physiology ; Cell Cycle - physiology ; Cell Proliferation ; Gene Expression Regulation, Neoplastic - genetics ; Glioblastoma - pathology ; Humans ; Laboratory Investigation - Human/Animal Tissue ; Medicine ; Medicine & Public Health ; Models, Animal ; Neurology ; Oncology ; Receptor, Epidermal Growth Factor - genetics ; Receptor, Epidermal Growth Factor - metabolism ; RNA, Messenger - metabolism ; SOXB1 Transcription Factors - metabolism ; Spheroids, Cellular - pathology ; Time Factors ; Tumor Cells, Cultured</subject><ispartof>Journal of neuro-oncology, 2011-05, Vol.102 (3), p.395-407</ispartof><rights>The Author(s) 2010</rights><rights>Springer Science+Business Media, LLC. 2011</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c468t-78bbb64203dd355fa320690c2351e9fa44c5c94e418c2b7e86433d28dc0895743</citedby><cites>FETCH-LOGICAL-c468t-78bbb64203dd355fa320690c2351e9fa44c5c94e418c2b7e86433d28dc0895743</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,315,786,790,891,27957,27958</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20803305$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Witusik-Perkowska, Monika</creatorcontrib><creatorcontrib>Rieske, Piotr</creatorcontrib><creatorcontrib>Hułas-Bigoszewska, Krystyna</creatorcontrib><creatorcontrib>Zakrzewska, Magdalena</creatorcontrib><creatorcontrib>Stawski, Robert</creatorcontrib><creatorcontrib>Kulczycka-Wojdala, Dominika</creatorcontrib><creatorcontrib>Bieńkowski, Michał</creatorcontrib><creatorcontrib>Stoczyńska-Fidelus, Ewelina</creatorcontrib><creatorcontrib>Grešner, Sylwia M.</creatorcontrib><creatorcontrib>Piaskowski, Sylwester</creatorcontrib><creatorcontrib>Jaskólski, Dariusz J.</creatorcontrib><creatorcontrib>Papierz, Wielisław</creatorcontrib><creatorcontrib>Zakrzewski, Krzysztof</creatorcontrib><creatorcontrib>Kolasa, Maciej</creatorcontrib><creatorcontrib>Ironside, James W.</creatorcontrib><creatorcontrib>Liberski, Paweł P.</creatorcontrib><title>Glioblastoma-derived spheroid cultures as an experimental model for analysis of EGFR anomalies</title><title>Journal of neuro-oncology</title><addtitle>J Neurooncol</addtitle><addtitle>J Neurooncol</addtitle><description>Glioblastoma cell cultures in vitro are frequently used for investigations on the biology of tumors or new therapeutic approaches. Recent reports have emphasized the importance of cell culture type for maintenance of tumor original features. Nevertheless, the ability of GBM cells to preserve EGFR overdosage in vitro remains controversial. Our experimental approach was based on quantitative analysis of
EGFR
gene dosage in vitro both at DNA and mRNA level. Real-time PCR data were verified with a FISH method allowing for a distinction between
EGFR
amplification and polysomy 7. We demonstrated that
EGFR
amplification accompanied by
EGFRwt
overexpression was maintained in spheroids, but these phenomena were gradually lost in adherent culture. We noticed a rapid decrease of
EGFR
overdosage already at the initial stage of cell culture establishment. In contrast to
EGFR
amplification, the maintenance of polysomy 7 resulted in
EGFR
locus gain and stabilization even in long-term adherent culture in serum presence. Surprisingly, the
EGFRwt
expression pattern did not reflect the latter phenomenon and we observed no overexpression of the tested gene. Moreover, quantitative analysis demonstrated that expression of the truncated variant of receptor—
EGFRvIII
was preserved in GBM-derived spheroids at a level comparable to the initial tumor tissue. Our findings are especially important in the light of research using glioblastoma culture as the experimental model for testing novel EGFR-targeted therapeutics in vitro, with special emphasis on the most common mutated form of receptor—EGFRvIII.</description><subject>Animals</subject><subject>Brain Neoplasms - pathology</subject><subject>Bromodeoxyuridine - metabolism</subject><subject>Cell Adhesion - physiology</subject><subject>Cell Cycle - physiology</subject><subject>Cell Proliferation</subject><subject>Gene Expression Regulation, Neoplastic - genetics</subject><subject>Glioblastoma - pathology</subject><subject>Humans</subject><subject>Laboratory Investigation - Human/Animal Tissue</subject><subject>Medicine</subject><subject>Medicine & Public Health</subject><subject>Models, Animal</subject><subject>Neurology</subject><subject>Oncology</subject><subject>Receptor, Epidermal Growth Factor - genetics</subject><subject>Receptor, Epidermal Growth Factor - metabolism</subject><subject>RNA, Messenger - metabolism</subject><subject>SOXB1 Transcription Factors - metabolism</subject><subject>Spheroids, Cellular - pathology</subject><subject>Time Factors</subject><subject>Tumor Cells, Cultured</subject><issn>0167-594X</issn><issn>1573-7373</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><recordid>eNp1kUtLAzEUhYMoWqs_wI0M7kdvXpOZjSClVqEgiIIrQybJtCNpU5MZ0X9vpLXqQsgDcs_97gkHoRMM5xhAXESMoYAccNqUkxx20ABzQXNBBd1FA8CFyHnFng7QYYwvAMAExfvogEAJlAIfoOeJa33tVOz8QuXGhvbNmiyu5jb41mS6d10fbMxUWsvMvq-SYmGXnXLZwhvrssaHVFHuI7Yx8002nlzfp4dEc62NR2ivUS7a4809RI_X44fRTT69m9yOrqa5ZkXZ5aKs67pgBKgxlPNGUQJFBZpQjm3VKMY01xWzDJea1MKWBaPUkNJoKCsuGB2iyzV31dcLa3RyGJSTq2RWhQ_pVSv_VpbtXM78m6QJIAhOgLMNIPjX3sZOvvg-pH9FWRYcKsxIlUR4LdLBxxhssx2AQX4lIteJyJSI_EokHUN0-tvZtuM7giQga0FMpeXMhp_J_1M_AbAql1w</recordid><startdate>20110501</startdate><enddate>20110501</enddate><creator>Witusik-Perkowska, Monika</creator><creator>Rieske, Piotr</creator><creator>Hułas-Bigoszewska, Krystyna</creator><creator>Zakrzewska, Magdalena</creator><creator>Stawski, Robert</creator><creator>Kulczycka-Wojdala, Dominika</creator><creator>Bieńkowski, Michał</creator><creator>Stoczyńska-Fidelus, Ewelina</creator><creator>Grešner, Sylwia M.</creator><creator>Piaskowski, Sylwester</creator><creator>Jaskólski, Dariusz J.</creator><creator>Papierz, Wielisław</creator><creator>Zakrzewski, Krzysztof</creator><creator>Kolasa, Maciej</creator><creator>Ironside, James W.</creator><creator>Liberski, Paweł P.</creator><general>Springer US</general><general>Springer Nature B.V</general><scope>C6C</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>5PM</scope></search><sort><creationdate>20110501</creationdate><title>Glioblastoma-derived spheroid cultures as an experimental model for analysis of EGFR anomalies</title><author>Witusik-Perkowska, Monika ; Rieske, Piotr ; Hułas-Bigoszewska, Krystyna ; Zakrzewska, Magdalena ; Stawski, Robert ; Kulczycka-Wojdala, Dominika ; Bieńkowski, Michał ; Stoczyńska-Fidelus, Ewelina ; Grešner, Sylwia M. ; Piaskowski, Sylwester ; Jaskólski, Dariusz J. ; Papierz, Wielisław ; Zakrzewski, Krzysztof ; Kolasa, Maciej ; Ironside, James W. ; Liberski, Paweł P.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c468t-78bbb64203dd355fa320690c2351e9fa44c5c94e418c2b7e86433d28dc0895743</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animals</topic><topic>Brain Neoplasms - 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Recent reports have emphasized the importance of cell culture type for maintenance of tumor original features. Nevertheless, the ability of GBM cells to preserve EGFR overdosage in vitro remains controversial. Our experimental approach was based on quantitative analysis of
EGFR
gene dosage in vitro both at DNA and mRNA level. Real-time PCR data were verified with a FISH method allowing for a distinction between
EGFR
amplification and polysomy 7. We demonstrated that
EGFR
amplification accompanied by
EGFRwt
overexpression was maintained in spheroids, but these phenomena were gradually lost in adherent culture. We noticed a rapid decrease of
EGFR
overdosage already at the initial stage of cell culture establishment. In contrast to
EGFR
amplification, the maintenance of polysomy 7 resulted in
EGFR
locus gain and stabilization even in long-term adherent culture in serum presence. Surprisingly, the
EGFRwt
expression pattern did not reflect the latter phenomenon and we observed no overexpression of the tested gene. Moreover, quantitative analysis demonstrated that expression of the truncated variant of receptor—
EGFRvIII
was preserved in GBM-derived spheroids at a level comparable to the initial tumor tissue. Our findings are especially important in the light of research using glioblastoma culture as the experimental model for testing novel EGFR-targeted therapeutics in vitro, with special emphasis on the most common mutated form of receptor—EGFRvIII.</abstract><cop>Boston</cop><pub>Springer US</pub><pmid>20803305</pmid><doi>10.1007/s11060-010-0352-0</doi><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Journal of neuro-oncology, 2011-05, Vol.102 (3), p.395-407 |
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| subjects | Animals Brain Neoplasms - pathology Bromodeoxyuridine - metabolism Cell Adhesion - physiology Cell Cycle - physiology Cell Proliferation Gene Expression Regulation, Neoplastic - genetics Glioblastoma - pathology Humans Laboratory Investigation - Human/Animal Tissue Medicine Medicine & Public Health Models, Animal Neurology Oncology Receptor, Epidermal Growth Factor - genetics Receptor, Epidermal Growth Factor - metabolism RNA, Messenger - metabolism SOXB1 Transcription Factors - metabolism Spheroids, Cellular - pathology Time Factors Tumor Cells, Cultured |
| title | Glioblastoma-derived spheroid cultures as an experimental model for analysis of EGFR anomalies |
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