Messenger RNA expression profiling of genes involved in epidermal growth factor receptor signalling in human cancer cells treated with scanning array-designed antisense oligonucleotides.
Identifieur interne : 000212 ( Ncbi/Merge ); précédent : 000211; suivant : 000213Messenger RNA expression profiling of genes involved in epidermal growth factor receptor signalling in human cancer cells treated with scanning array-designed antisense oligonucleotides.
Auteurs : Amelia K. Petch [Royaume-Uni] ; Muhammad Sohail ; Marcus D. Hughes ; Ibrahim Benter ; John Darling ; Edwin M. Southern ; Saghir AkhtarSource :
- Biochemical pharmacology [ 0006-2952 ] ; 2003.
Descripteurs français
- KwdFr :
- ARN messager (), ARN messager (métabolisme), Analyse de profil d'expression de gènes, Cellules cancéreuses en culture, Division cellulaire (), Gliome (anatomopathologie), Humains, Oligonucléotides antisens (pharmacologie), Ribonuclease H (métabolisme), Récepteurs ErbB (génétique), Récepteurs ErbB (métabolisme), Régulation de l'expression des gènes tumoraux (), Régulation négative, Séquençage par oligonucléotides en batterie, Transduction du signal.
- MESH :
- anatomopathologie : Gliome.
- génétique : Récepteurs ErbB.
- métabolisme : ARN messager, Ribonuclease H, Récepteurs ErbB.
- pharmacologie : Oligonucléotides antisens.
- ARN messager, Analyse de profil d'expression de gènes, Cellules cancéreuses en culture, Division cellulaire, Humains, Régulation de l'expression des gènes tumoraux, Régulation négative, Séquençage par oligonucléotides en batterie, Transduction du signal.
English descriptors
- KwdEn :
- Cell Division (drug effects), Down-Regulation, ErbB Receptors (genetics), ErbB Receptors (metabolism), Gene Expression Profiling, Gene Expression Regulation, Neoplastic (drug effects), Glioma (pathology), Humans, Oligonucleotide Array Sequence Analysis, Oligonucleotides, Antisense (pharmacology), RNA, Messenger (drug effects), RNA, Messenger (metabolism), Ribonuclease H (metabolism), Signal Transduction, Tumor Cells, Cultured.
- MESH :
- chemical , drug effects : RNA, Messenger.
- chemical , genetics : ErbB Receptors.
- drug effects : Cell Division, Gene Expression Regulation, Neoplastic.
- chemical , metabolism : ErbB Receptors, RNA, Messenger, Ribonuclease H.
- pathology : Glioma.
- chemical , pharmacology : Oligonucleotides, Antisense.
- Down-Regulation, Gene Expression Profiling, Humans, Oligonucleotide Array Sequence Analysis, Signal Transduction, Tumor Cells, Cultured.
Abstract
Scanning oligodeoxynucleotide (ODN) arrays appear promising in vitro tools for the prediction of effective antisense reagents but their usefulness has not yet been reported in mammalian systems. In this study, we have evaluated the use of scanning ODN arrays to predict efficacious antisense ODNs targeting the human epidermal growth factor receptor (EGFR) mRNA in a human epidermoid cancer cell line and in primary human glioma cells. Hybridisation accessibility profile of the first 120nt in the coding region of the human EGFR mRNA was determined by hybridising a radiolabelled EGFR transcript to a scanning array of 2684 antisense sequences ranging from monomers to 27-mers. Two ODNs, AS1 and AS2, complementary to accessible sequences within the EGFR mRNA, were designed and their ability to hybridise to EGFR mRNA was further confirmed by in vitro RNase H-mediated cleavage assays. Phosphorothioate-modified 21-mer AS1 and AS2 ODNs inhibited the growth of an established human A431 cancer cell line as well as primary glioma cells from human subjects when delivered as cationic lipoplexes. In contrast, scrambled controls and AS3-an antisense ODN complementary to an inaccessible site in EGFR mRNA-were inactive. Western blots showed that AS1 ODN exhibited a dose-dependent inhibition of EGFR protein expression in A431 cells in the nanomolar range. Microarray-based gene expression profiling studies of A431 cells treated with the 21-mer phosphorothioate AS1 ODN demonstrated successful inhibition of downstream signalling molecules further confirming the effective inhibition of EGFR expression in human cancer cells by antisense ODNs designed by scanning ODN array technology.
DOI: 10.1016/s0006-2952(03)00407-6
PubMed: 12948863
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pubmed:12948863Le document en format XML
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Petch</name><affiliation wicri:level="1"><nlm:affiliation>Pharmaceutical Sciences Research Institute, Aston University, Aston Triangle, Birmingham B4 7ET, UK.</nlm:affiliation><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Pharmaceutical Sciences Research Institute, Aston University, Aston Triangle, Birmingham B4 7ET</wicri:regionArea><wicri:noRegion>Birmingham B4 7ET</wicri:noRegion></affiliation></author><author><name sortKey="Sohail, Muhammad" sort="Sohail, Muhammad" uniqKey="Sohail M" first="Muhammad" last="Sohail">Muhammad Sohail</name></author><author><name sortKey="Hughes, Marcus D" sort="Hughes, Marcus D" uniqKey="Hughes M" first="Marcus D" last="Hughes">Marcus D. 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In this study, we have evaluated the use of scanning ODN arrays to predict efficacious antisense ODNs targeting the human epidermal growth factor receptor (EGFR) mRNA in a human epidermoid cancer cell line and in primary human glioma cells. Hybridisation accessibility profile of the first 120nt in the coding region of the human EGFR mRNA was determined by hybridising a radiolabelled EGFR transcript to a scanning array of 2684 antisense sequences ranging from monomers to 27-mers. Two ODNs, AS1 and AS2, complementary to accessible sequences within the EGFR mRNA, were designed and their ability to hybridise to EGFR mRNA was further confirmed by in vitro RNase H-mediated cleavage assays. Phosphorothioate-modified 21-mer AS1 and AS2 ODNs inhibited the growth of an established human A431 cancer cell line as well as primary glioma cells from human subjects when delivered as cationic lipoplexes. In contrast, scrambled controls and AS3-an antisense ODN complementary to an inaccessible site in EGFR mRNA-were inactive. Western blots showed that AS1 ODN exhibited a dose-dependent inhibition of EGFR protein expression in A431 cells in the nanomolar range. Microarray-based gene expression profiling studies of A431 cells treated with the 21-mer phosphorothioate AS1 ODN demonstrated successful inhibition of downstream signalling molecules further confirming the effective inhibition of EGFR expression in human cancer cells by antisense ODNs designed by scanning ODN array technology.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">12948863</PMID><DateCompleted><Year>2003</Year><Month>10</Month><Day>06</Day></DateCompleted><DateRevised><Year>2019</Year><Month>06</Month><Day>23</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0006-2952</ISSN><JournalIssue CitedMedium="Print"><Volume>66</Volume><Issue>5</Issue><PubDate><Year>2003</Year><Month>Sep</Month><Day>01</Day></PubDate></JournalIssue><Title>Biochemical pharmacology</Title><ISOAbbreviation>Biochem. Pharmacol.</ISOAbbreviation></Journal><ArticleTitle>Messenger RNA expression profiling of genes involved in epidermal growth factor receptor signalling in human cancer cells treated with scanning array-designed antisense oligonucleotides.</ArticleTitle><Pagination><MedlinePgn>819-30</MedlinePgn></Pagination><Abstract><AbstractText>Scanning oligodeoxynucleotide (ODN) arrays appear promising in vitro tools for the prediction of effective antisense reagents but their usefulness has not yet been reported in mammalian systems. In this study, we have evaluated the use of scanning ODN arrays to predict efficacious antisense ODNs targeting the human epidermal growth factor receptor (EGFR) mRNA in a human epidermoid cancer cell line and in primary human glioma cells. Hybridisation accessibility profile of the first 120nt in the coding region of the human EGFR mRNA was determined by hybridising a radiolabelled EGFR transcript to a scanning array of 2684 antisense sequences ranging from monomers to 27-mers. Two ODNs, AS1 and AS2, complementary to accessible sequences within the EGFR mRNA, were designed and their ability to hybridise to EGFR mRNA was further confirmed by in vitro RNase H-mediated cleavage assays. Phosphorothioate-modified 21-mer AS1 and AS2 ODNs inhibited the growth of an established human A431 cancer cell line as well as primary glioma cells from human subjects when delivered as cationic lipoplexes. In contrast, scrambled controls and AS3-an antisense ODN complementary to an inaccessible site in EGFR mRNA-were inactive. Western blots showed that AS1 ODN exhibited a dose-dependent inhibition of EGFR protein expression in A431 cells in the nanomolar range. Microarray-based gene expression profiling studies of A431 cells treated with the 21-mer phosphorothioate AS1 ODN demonstrated successful inhibition of downstream signalling molecules further confirming the effective inhibition of EGFR expression in human cancer cells by antisense ODNs designed by scanning ODN array technology.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Petch</LastName><ForeName>Amelia K</ForeName><Initials>AK</Initials><AffiliationInfo><Affiliation>Pharmaceutical Sciences Research Institute, Aston University, Aston Triangle, Birmingham B4 7ET, UK.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Sohail</LastName><ForeName>Muhammad</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Hughes</LastName><ForeName>Marcus D</ForeName><Initials>MD</Initials></Author><Author ValidYN="Y"><LastName>Benter</LastName><ForeName>Ibrahim</ForeName><Initials>I</Initials></Author><Author ValidYN="Y"><LastName>Darling</LastName><ForeName>John</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Southern</LastName><ForeName>Edwin M</ForeName><Initials>EM</Initials></Author><Author ValidYN="Y"><LastName>Akhtar</LastName><ForeName>Saghir</ForeName><Initials>S</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Biochem Pharmacol</MedlineTA><NlmUniqueID>0101032</NlmUniqueID><ISSNLinking>0006-2952</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D016376">Oligonucleotides, Antisense</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D012333">RNA, 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Profiling</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015972" MajorTopicYN="N">Gene Expression Regulation, Neoplastic</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="Y">drug effects</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005910" MajorTopicYN="N">Glioma</DescriptorName><QualifierName UI="Q000473" MajorTopicYN="N">pathology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020411" MajorTopicYN="N">Oligonucleotide Array Sequence Analysis</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D016376" MajorTopicYN="N">Oligonucleotides, Antisense</DescriptorName><QualifierName UI="Q000494" MajorTopicYN="Y">pharmacology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012333" MajorTopicYN="N">RNA, Messenger</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D016914" MajorTopicYN="N">Ribonuclease H</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015398" MajorTopicYN="N">Signal Transduction</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014407" MajorTopicYN="N">Tumor Cells, Cultured</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2003</Year><Month>9</Month><Day>2</Day><Hour>5</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2003</Year><Month>10</Month><Day>8</Day><Hour>5</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2003</Year><Month>9</Month><Day>2</Day><Hour>5</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">12948863</ArticleId><ArticleId IdType="pii">S0006295203004076</ArticleId><ArticleId IdType="doi">10.1016/s0006-2952(03)00407-6</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Royaume-Uni</li></country></list><tree><noCountry><name sortKey="Akhtar, Saghir" sort="Akhtar, Saghir" uniqKey="Akhtar S" first="Saghir" last="Akhtar">Saghir Akhtar</name><name sortKey="Benter, Ibrahim" sort="Benter, Ibrahim" uniqKey="Benter I" first="Ibrahim" last="Benter">Ibrahim Benter</name><name sortKey="Darling, John" sort="Darling, John" uniqKey="Darling J" first="John" last="Darling">John Darling</name><name sortKey="Hughes, Marcus D" sort="Hughes, Marcus D" uniqKey="Hughes M" first="Marcus D" last="Hughes">Marcus D. Hughes</name><name sortKey="Sohail, Muhammad" sort="Sohail, Muhammad" uniqKey="Sohail M" first="Muhammad" last="Sohail">Muhammad Sohail</name><name sortKey="Southern, Edwin M" sort="Southern, Edwin M" uniqKey="Southern E" first="Edwin M" last="Southern">Edwin M. Southern</name></noCountry><country name="Royaume-Uni"><noRegion><name sortKey="Petch, Amelia K" sort="Petch, Amelia K" uniqKey="Petch A" first="Amelia K" last="Petch">Amelia K. Petch</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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