Identification of differentially expressed proteins in human glioblastoma cell lines and tumors
An in‐frame deletion of 801 bp in exons 2–7 (type III mutation) of the epidermal growth factor receptor (EGFR) is detected at high incidence in primary glioblastoma tumors. A proteomic approach was used to generate differential protein expression maps of fetal human astrocytes (FHA), human glioblast...
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Veröffentlicht in: | Glia 2003-04, Vol.42 (2), p.194-208 |
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description | An in‐frame deletion of 801 bp in exons 2–7 (type III mutation) of the epidermal growth factor receptor (EGFR) is detected at high incidence in primary glioblastoma tumors. A proteomic approach was used to generate differential protein expression maps of fetal human astrocytes (FHA), human glioblastoma cell lines U87MG and U87MG expressing type III EGFR deletion (U87MGΔEGFR) that confers high malignancy to tumor cells. Two‐dimensional gel electrophoresis followed by in‐gel digestion of separated spots and protein identification by LC‐MS‐MS and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) identified 23 proteins expressed at higher levels or exclusively in FHA and 29 proteins expressed at higher levels or exclusively in U87MG cells. Three proteins, ubiquitin, cystatin B, and tissue transglutaminase (TTG), were upregulated in U87MGΔEGFR relative to U87MG. Four proteins highly expressed by U87MG cells, Hsp27, major vault protein, TTG, and cystatin B, were analyzed by Western blot, ELISA, or RT‐PCR in cell extracts and in tissue samples of glioblastoma multiforme (GBM; grade IV), low‐grade astrocytomas (grades I and II), and nonmalignant brain lesions. All four proteins were highly expressed in GBM tissues compared to nonmalignant brain. These proteins may be used as diagnostic or functional (e.g., multiple drug resistance, invasiveness) markers for glioblastoma tumors. GLIA 42:194–208, 2003. © 2003 Wiley‐Liss, Inc. |
doi_str_mv | 10.1002/glia.10222 |
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A proteomic approach was used to generate differential protein expression maps of fetal human astrocytes (FHA), human glioblastoma cell lines U87MG and U87MG expressing type III EGFR deletion (U87MGΔEGFR) that confers high malignancy to tumor cells. Two‐dimensional gel electrophoresis followed by in‐gel digestion of separated spots and protein identification by LC‐MS‐MS and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) identified 23 proteins expressed at higher levels or exclusively in FHA and 29 proteins expressed at higher levels or exclusively in U87MG cells. Three proteins, ubiquitin, cystatin B, and tissue transglutaminase (TTG), were upregulated in U87MGΔEGFR relative to U87MG. Four proteins highly expressed by U87MG cells, Hsp27, major vault protein, TTG, and cystatin B, were analyzed by Western blot, ELISA, or RT‐PCR in cell extracts and in tissue samples of glioblastoma multiforme (GBM; grade IV), low‐grade astrocytomas (grades I and II), and nonmalignant brain lesions. All four proteins were highly expressed in GBM tissues compared to nonmalignant brain. These proteins may be used as diagnostic or functional (e.g., multiple drug resistance, invasiveness) markers for glioblastoma tumors. GLIA 42:194–208, 2003. © 2003 Wiley‐Liss, Inc.</description><identifier>ISSN: 0894-1491</identifier><identifier>EISSN: 1098-1136</identifier><identifier>DOI: 10.1002/glia.10222</identifier><identifier>PMID: 12655603</identifier><identifier>CODEN: GLIAEJ</identifier><language>eng</language><publisher>New York: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Astrocytes - metabolism ; Biological and medical sciences ; Biomarkers, Tumor - metabolism ; Brain Neoplasms - diagnosis ; Brain Neoplasms - genetics ; Brain Neoplasms - metabolism ; Cystatin B ; Cystatins - metabolism ; Electrophoresis, Gel, Two-Dimensional ; Epidermal Growth Factor - genetics ; epidermal growth factor receptor ; Fetus ; Gene Expression Regulation, Neoplastic - physiology ; glioblastoma ; Glioblastoma - diagnosis ; Glioblastoma - genetics ; Glioblastoma - metabolism ; Heat-Shock Proteins ; Hsp27 ; HSP27 Heat-Shock Proteins ; Humans ; liquid chromatography tandem mass spectrometry (LC-MS-MS) ; major vault protein ; matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) ; Medical sciences ; Mutation - genetics ; Neoplasm Proteins - metabolism ; Neurology ; Phosphorylation ; Proteins - metabolism ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; tissue transglutaminase ; Transglutaminases - metabolism ; Tumor Cells, Cultured - cytology ; Tumor Cells, Cultured - metabolism ; Tumors of the nervous system. Phacomatoses ; U87MG ; Ubiquitin - metabolism ; Up-Regulation - physiology ; Vault Ribonucleoprotein Particles - metabolism</subject><ispartof>Glia, 2003-04, Vol.42 (2), p.194-208</ispartof><rights>Copyright © 2003 Wiley‐Liss, Inc.</rights><rights>2003 INIST-CNRS</rights><rights>Copyright 2003 Wiley-Liss, Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5552-6edfa98eab8fce4e42ba65b1e0c58b5dc12be05ea0ea254e3449309f643e140d3</citedby><cites>FETCH-LOGICAL-c5552-6edfa98eab8fce4e42ba65b1e0c58b5dc12be05ea0ea254e3449309f643e140d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fglia.10222$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fglia.10222$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14714719$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12655603$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhang, Rulin</creatorcontrib><creatorcontrib>Tremblay, Tammy-Lynn</creatorcontrib><creatorcontrib>Mcdermid, Angela</creatorcontrib><creatorcontrib>Thibault, Pierre</creatorcontrib><creatorcontrib>Stanimirovic, Danica</creatorcontrib><title>Identification of differentially expressed proteins in human glioblastoma cell lines and tumors</title><title>Glia</title><addtitle>Glia</addtitle><description>An in‐frame deletion of 801 bp in exons 2–7 (type III mutation) of the epidermal growth factor receptor (EGFR) is detected at high incidence in primary glioblastoma tumors. A proteomic approach was used to generate differential protein expression maps of fetal human astrocytes (FHA), human glioblastoma cell lines U87MG and U87MG expressing type III EGFR deletion (U87MGΔEGFR) that confers high malignancy to tumor cells. Two‐dimensional gel electrophoresis followed by in‐gel digestion of separated spots and protein identification by LC‐MS‐MS and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) identified 23 proteins expressed at higher levels or exclusively in FHA and 29 proteins expressed at higher levels or exclusively in U87MG cells. Three proteins, ubiquitin, cystatin B, and tissue transglutaminase (TTG), were upregulated in U87MGΔEGFR relative to U87MG. Four proteins highly expressed by U87MG cells, Hsp27, major vault protein, TTG, and cystatin B, were analyzed by Western blot, ELISA, or RT‐PCR in cell extracts and in tissue samples of glioblastoma multiforme (GBM; grade IV), low‐grade astrocytomas (grades I and II), and nonmalignant brain lesions. All four proteins were highly expressed in GBM tissues compared to nonmalignant brain. These proteins may be used as diagnostic or functional (e.g., multiple drug resistance, invasiveness) markers for glioblastoma tumors. GLIA 42:194–208, 2003. © 2003 Wiley‐Liss, Inc.</description><subject>Astrocytes - metabolism</subject><subject>Biological and medical sciences</subject><subject>Biomarkers, Tumor - metabolism</subject><subject>Brain Neoplasms - diagnosis</subject><subject>Brain Neoplasms - genetics</subject><subject>Brain Neoplasms - metabolism</subject><subject>Cystatin B</subject><subject>Cystatins - metabolism</subject><subject>Electrophoresis, Gel, Two-Dimensional</subject><subject>Epidermal Growth Factor - genetics</subject><subject>epidermal growth factor receptor</subject><subject>Fetus</subject><subject>Gene Expression Regulation, Neoplastic - physiology</subject><subject>glioblastoma</subject><subject>Glioblastoma - diagnosis</subject><subject>Glioblastoma - genetics</subject><subject>Glioblastoma - metabolism</subject><subject>Heat-Shock Proteins</subject><subject>Hsp27</subject><subject>HSP27 Heat-Shock Proteins</subject><subject>Humans</subject><subject>liquid chromatography tandem mass spectrometry (LC-MS-MS)</subject><subject>major vault protein</subject><subject>matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF)</subject><subject>Medical sciences</subject><subject>Mutation - genetics</subject><subject>Neoplasm Proteins - metabolism</subject><subject>Neurology</subject><subject>Phosphorylation</subject><subject>Proteins - metabolism</subject><subject>Proteomics</subject><subject>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</subject><subject>tissue transglutaminase</subject><subject>Transglutaminases - metabolism</subject><subject>Tumor Cells, Cultured - cytology</subject><subject>Tumor Cells, Cultured - metabolism</subject><subject>Tumors of the nervous system. Phacomatoses</subject><subject>U87MG</subject><subject>Ubiquitin - metabolism</subject><subject>Up-Regulation - physiology</subject><subject>Vault Ribonucleoprotein Particles - metabolism</subject><issn>0894-1491</issn><issn>1098-1136</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1v1DAQhi0EosvChR-AfIEDUlp_5uNYFVgWrdpLEUdr4ozB4DiLnYjuv2_CLvQG0kiesZ533tFLyEvOzjlj4uJr8DB3QohHZMVZUxecy_IxWbG6UQVXDT8jz3L-zhifh-opOeOi1LpkckXMtsM4euctjH6IdHC0885hWn4hhAPFu33CnLGj-zSM6GOmPtJvUw-Rzs5DGyCPQw_UYgg0-IiZQuzoOPVDys_JEwch44vTuyafP7y_vfpY7G4226vLXWG11qIosXPQ1Aht7SwqVKKFUrccmdV1qzvLRYtMIzAEoRVKpRrJGlcqiVyxTq7Jm-Pe-cifE-bR9D4vF0HEYcqmklzOEv5fULBS1XpG1-TtEbRpyDmhM_vke0gHw5lZcjdL7uZ37jP86rR1anvsHtBT0DPw-gRAthBcgmh9fuBUtdTiyo_cLx_w8A9Ls9ltL_-YF0eNzyPe_dVA-mHKSlbafLnemOtN847z3a35JO8BC5OrFQ</recordid><startdate>20030415</startdate><enddate>20030415</enddate><creator>Zhang, Rulin</creator><creator>Tremblay, Tammy-Lynn</creator><creator>Mcdermid, Angela</creator><creator>Thibault, Pierre</creator><creator>Stanimirovic, Danica</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley-Liss</general><scope>BSCLL</scope><scope>IQODW</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>7TK</scope><scope>7X8</scope></search><sort><creationdate>20030415</creationdate><title>Identification of differentially expressed proteins in human glioblastoma cell lines and tumors</title><author>Zhang, Rulin ; Tremblay, Tammy-Lynn ; Mcdermid, Angela ; Thibault, Pierre ; Stanimirovic, Danica</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5552-6edfa98eab8fce4e42ba65b1e0c58b5dc12be05ea0ea254e3449309f643e140d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Astrocytes - metabolism</topic><topic>Biological and medical sciences</topic><topic>Biomarkers, Tumor - metabolism</topic><topic>Brain Neoplasms - diagnosis</topic><topic>Brain Neoplasms - genetics</topic><topic>Brain Neoplasms - metabolism</topic><topic>Cystatin B</topic><topic>Cystatins - metabolism</topic><topic>Electrophoresis, Gel, Two-Dimensional</topic><topic>Epidermal Growth Factor - genetics</topic><topic>epidermal growth factor receptor</topic><topic>Fetus</topic><topic>Gene Expression Regulation, Neoplastic - physiology</topic><topic>glioblastoma</topic><topic>Glioblastoma - diagnosis</topic><topic>Glioblastoma - genetics</topic><topic>Glioblastoma - metabolism</topic><topic>Heat-Shock Proteins</topic><topic>Hsp27</topic><topic>HSP27 Heat-Shock Proteins</topic><topic>Humans</topic><topic>liquid chromatography tandem mass spectrometry (LC-MS-MS)</topic><topic>major vault protein</topic><topic>matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF)</topic><topic>Medical sciences</topic><topic>Mutation - genetics</topic><topic>Neoplasm Proteins - metabolism</topic><topic>Neurology</topic><topic>Phosphorylation</topic><topic>Proteins - metabolism</topic><topic>Proteomics</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</topic><topic>tissue transglutaminase</topic><topic>Transglutaminases - metabolism</topic><topic>Tumor Cells, Cultured - cytology</topic><topic>Tumor Cells, Cultured - metabolism</topic><topic>Tumors of the nervous system. Phacomatoses</topic><topic>U87MG</topic><topic>Ubiquitin - metabolism</topic><topic>Up-Regulation - physiology</topic><topic>Vault Ribonucleoprotein Particles - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhang, Rulin</creatorcontrib><creatorcontrib>Tremblay, Tammy-Lynn</creatorcontrib><creatorcontrib>Mcdermid, Angela</creatorcontrib><creatorcontrib>Thibault, Pierre</creatorcontrib><creatorcontrib>Stanimirovic, Danica</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Neurosciences Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Glia</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhang, Rulin</au><au>Tremblay, Tammy-Lynn</au><au>Mcdermid, Angela</au><au>Thibault, Pierre</au><au>Stanimirovic, Danica</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of differentially expressed proteins in human glioblastoma cell lines and tumors</atitle><jtitle>Glia</jtitle><addtitle>Glia</addtitle><date>2003-04-15</date><risdate>2003</risdate><volume>42</volume><issue>2</issue><spage>194</spage><epage>208</epage><pages>194-208</pages><issn>0894-1491</issn><eissn>1098-1136</eissn><coden>GLIAEJ</coden><abstract>An in‐frame deletion of 801 bp in exons 2–7 (type III mutation) of the epidermal growth factor receptor (EGFR) is detected at high incidence in primary glioblastoma tumors. A proteomic approach was used to generate differential protein expression maps of fetal human astrocytes (FHA), human glioblastoma cell lines U87MG and U87MG expressing type III EGFR deletion (U87MGΔEGFR) that confers high malignancy to tumor cells. Two‐dimensional gel electrophoresis followed by in‐gel digestion of separated spots and protein identification by LC‐MS‐MS and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) identified 23 proteins expressed at higher levels or exclusively in FHA and 29 proteins expressed at higher levels or exclusively in U87MG cells. Three proteins, ubiquitin, cystatin B, and tissue transglutaminase (TTG), were upregulated in U87MGΔEGFR relative to U87MG. Four proteins highly expressed by U87MG cells, Hsp27, major vault protein, TTG, and cystatin B, were analyzed by Western blot, ELISA, or RT‐PCR in cell extracts and in tissue samples of glioblastoma multiforme (GBM; grade IV), low‐grade astrocytomas (grades I and II), and nonmalignant brain lesions. All four proteins were highly expressed in GBM tissues compared to nonmalignant brain. These proteins may be used as diagnostic or functional (e.g., multiple drug resistance, invasiveness) markers for glioblastoma tumors. GLIA 42:194–208, 2003. © 2003 Wiley‐Liss, Inc.</abstract><cop>New York</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>12655603</pmid><doi>10.1002/glia.10222</doi><tpages>15</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Astrocytes - metabolism Biological and medical sciences Biomarkers, Tumor - metabolism Brain Neoplasms - diagnosis Brain Neoplasms - genetics Brain Neoplasms - metabolism Cystatin B Cystatins - metabolism Electrophoresis, Gel, Two-Dimensional Epidermal Growth Factor - genetics epidermal growth factor receptor Fetus Gene Expression Regulation, Neoplastic - physiology glioblastoma Glioblastoma - diagnosis Glioblastoma - genetics Glioblastoma - metabolism Heat-Shock Proteins Hsp27 HSP27 Heat-Shock Proteins Humans liquid chromatography tandem mass spectrometry (LC-MS-MS) major vault protein matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) Medical sciences Mutation - genetics Neoplasm Proteins - metabolism Neurology Phosphorylation Proteins - metabolism Proteomics Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization tissue transglutaminase Transglutaminases - metabolism Tumor Cells, Cultured - cytology Tumor Cells, Cultured - metabolism Tumors of the nervous system. Phacomatoses U87MG Ubiquitin - metabolism Up-Regulation - physiology Vault Ribonucleoprotein Particles - metabolism |
title | Identification of differentially expressed proteins in human glioblastoma cell lines and tumors |
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