Selection of suitable housekeeping genes for expression analysis in glioblastoma using quantitative RT-PCR
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/31378 http://dx.doi.org/10.1186/1471-2199-10-17 |
Resumo: | Background: Considering the broad variation in the expression of housekeeping genes among tissues and experimental situations, studies using quantitative RT-PCR require strict definition of adequate endogenous controls. for glioblastoma, the most common type of tumor in the central nervous system, there was no previous report regarding this issue.Results: Here we show that amongst seven frequently used housekeeping genes TBP and HPRT1 are adequate references for glioblastoma gene expression analysis. Evaluation of the expression levels of 12 target genes utilizing different endogenous controls revealed that the normalization method applied might introduce errors in the estimation of relative quantities. Genes presenting expression levels which do not significantly differ between tumor and normal tissues can be considered either increased or decreased if unsuitable reference genes are applied. Most importantly, genes showing significant differences in expression levels between tumor and normal tissues can be missed. We also demonstrated that the Holliday Junction Recognizing Protein, a novel DNA repair protein over expressed in lung cancer, is extremely over-expressed in glioblastoma, with a median change of about 134 fold.Conclusion: Altogether, our data show the relevance of previous validation of candidate control genes for each experimental model and indicate TBP plus HPRT1 as suitable references for studies on glioblastoma gene expression. |
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Valente, ValeriaTeixeira, Silvia A.Neder, LucianoOkamoto, Oswaldo Keith [UNIFESP]Oba-Shinjo, Sueli Mieko [UNIFESP]Marie, Suely K. N.Scrideli, Carlos A.Paco-Larson, Maria L.Carlotti, Carlos G.Universidade de São Paulo (USP)Universidade Federal de São Paulo (UNIFESP)2016-01-24T13:52:21Z2016-01-24T13:52:21Z2009-03-03Bmc Molecular Biology. London: Biomed Central Ltd, v. 10, 11 p., 2009.1471-2199http://repositorio.unifesp.br/handle/11600/31378http://dx.doi.org/10.1186/1471-2199-10-17WOS000265057600001.pdf10.1186/1471-2199-10-17WOS:000265057600001Background: Considering the broad variation in the expression of housekeeping genes among tissues and experimental situations, studies using quantitative RT-PCR require strict definition of adequate endogenous controls. for glioblastoma, the most common type of tumor in the central nervous system, there was no previous report regarding this issue.Results: Here we show that amongst seven frequently used housekeeping genes TBP and HPRT1 are adequate references for glioblastoma gene expression analysis. Evaluation of the expression levels of 12 target genes utilizing different endogenous controls revealed that the normalization method applied might introduce errors in the estimation of relative quantities. Genes presenting expression levels which do not significantly differ between tumor and normal tissues can be considered either increased or decreased if unsuitable reference genes are applied. Most importantly, genes showing significant differences in expression levels between tumor and normal tissues can be missed. We also demonstrated that the Holliday Junction Recognizing Protein, a novel DNA repair protein over expressed in lung cancer, is extremely over-expressed in glioblastoma, with a median change of about 134 fold.Conclusion: Altogether, our data show the relevance of previous validation of candidate control genes for each experimental model and indicate TBP plus HPRT1 as suitable references for studies on glioblastoma gene expression.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAEPA-FMRUniv São Paulo, Fac Med, Dept Surg & Anat, BR-14049090 Ribeirao Preto, SP, BrazilUniv São Paulo, Fac Med, Dept Cellular & Mol Biol, BR-14049090 Ribeirao Preto, SP, BrazilUniv São Paulo, Fac Med, Dept Pathol, BR-14049090 Ribeirao Preto, SP, BrazilUniversidade Federal de São Paulo, Dept Neurol & Neurosurg, BR-04023900 São Paulo, BrazilUniv São Paulo, Sch Med, Dept Neurol, BR-01246903 São Paulo, BrazilUniv São Paulo, Fac Med, Dept Pediat, BR-14049090 Ribeirao Preto, SP, BrazilUniversidade Federal de São Paulo, Dept Neurol & Neurosurg, BR-04023900 São Paulo, BrazilFAPESP: 04/12133-6FAPESP: 06/57602-9CNPq: 485342/2006Web of Science11engBiomed Central LtdBmc Molecular BiologySelection of suitable housekeeping genes for expression analysis in glioblastoma using quantitative RT-PCRinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALWOS000265057600001.pdfapplication/pdf766128${dspace.ui.url}/bitstream/11600/31378/1/WOS000265057600001.pdfd5da1a804c67b45666ed8a3663b2bd78MD51open accessTEXTWOS000265057600001.pdf.txtWOS000265057600001.pdf.txtExtracted texttext/plain50341${dspace.ui.url}/bitstream/11600/31378/2/WOS000265057600001.pdf.txt0ee29983f18b3ab22560ca9a1210b30dMD52open access11600/313782023-01-30 22:17:46.939open accessoai:repositorio.unifesp.br:11600/31378Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-01-31T01:17:46Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Selection of suitable housekeeping genes for expression analysis in glioblastoma using quantitative RT-PCR |
title |
Selection of suitable housekeeping genes for expression analysis in glioblastoma using quantitative RT-PCR |
spellingShingle |
Selection of suitable housekeeping genes for expression analysis in glioblastoma using quantitative RT-PCR Valente, Valeria |
title_short |
Selection of suitable housekeeping genes for expression analysis in glioblastoma using quantitative RT-PCR |
title_full |
Selection of suitable housekeeping genes for expression analysis in glioblastoma using quantitative RT-PCR |
title_fullStr |
Selection of suitable housekeeping genes for expression analysis in glioblastoma using quantitative RT-PCR |
title_full_unstemmed |
Selection of suitable housekeeping genes for expression analysis in glioblastoma using quantitative RT-PCR |
title_sort |
Selection of suitable housekeeping genes for expression analysis in glioblastoma using quantitative RT-PCR |
author |
Valente, Valeria |
author_facet |
Valente, Valeria Teixeira, Silvia A. Neder, Luciano Okamoto, Oswaldo Keith [UNIFESP] Oba-Shinjo, Sueli Mieko [UNIFESP] Marie, Suely K. N. Scrideli, Carlos A. Paco-Larson, Maria L. Carlotti, Carlos G. |
author_role |
author |
author2 |
Teixeira, Silvia A. Neder, Luciano Okamoto, Oswaldo Keith [UNIFESP] Oba-Shinjo, Sueli Mieko [UNIFESP] Marie, Suely K. N. Scrideli, Carlos A. Paco-Larson, Maria L. Carlotti, Carlos G. |
author2_role |
author author author author author author author author |
dc.contributor.institution.none.fl_str_mv |
Universidade de São Paulo (USP) Universidade Federal de São Paulo (UNIFESP) |
dc.contributor.author.fl_str_mv |
Valente, Valeria Teixeira, Silvia A. Neder, Luciano Okamoto, Oswaldo Keith [UNIFESP] Oba-Shinjo, Sueli Mieko [UNIFESP] Marie, Suely K. N. Scrideli, Carlos A. Paco-Larson, Maria L. Carlotti, Carlos G. |
description |
Background: Considering the broad variation in the expression of housekeeping genes among tissues and experimental situations, studies using quantitative RT-PCR require strict definition of adequate endogenous controls. for glioblastoma, the most common type of tumor in the central nervous system, there was no previous report regarding this issue.Results: Here we show that amongst seven frequently used housekeeping genes TBP and HPRT1 are adequate references for glioblastoma gene expression analysis. Evaluation of the expression levels of 12 target genes utilizing different endogenous controls revealed that the normalization method applied might introduce errors in the estimation of relative quantities. Genes presenting expression levels which do not significantly differ between tumor and normal tissues can be considered either increased or decreased if unsuitable reference genes are applied. Most importantly, genes showing significant differences in expression levels between tumor and normal tissues can be missed. We also demonstrated that the Holliday Junction Recognizing Protein, a novel DNA repair protein over expressed in lung cancer, is extremely over-expressed in glioblastoma, with a median change of about 134 fold.Conclusion: Altogether, our data show the relevance of previous validation of candidate control genes for each experimental model and indicate TBP plus HPRT1 as suitable references for studies on glioblastoma gene expression. |
publishDate |
2009 |
dc.date.issued.fl_str_mv |
2009-03-03 |
dc.date.accessioned.fl_str_mv |
2016-01-24T13:52:21Z |
dc.date.available.fl_str_mv |
2016-01-24T13:52:21Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
Bmc Molecular Biology. London: Biomed Central Ltd, v. 10, 11 p., 2009. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/31378 http://dx.doi.org/10.1186/1471-2199-10-17 |
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1471-2199 |
dc.identifier.file.none.fl_str_mv |
WOS000265057600001.pdf |
dc.identifier.doi.none.fl_str_mv |
10.1186/1471-2199-10-17 |
dc.identifier.wos.none.fl_str_mv |
WOS:000265057600001 |
identifier_str_mv |
Bmc Molecular Biology. London: Biomed Central Ltd, v. 10, 11 p., 2009. 1471-2199 WOS000265057600001.pdf 10.1186/1471-2199-10-17 WOS:000265057600001 |
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http://repositorio.unifesp.br/handle/11600/31378 http://dx.doi.org/10.1186/1471-2199-10-17 |
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Biomed Central Ltd |
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Biomed Central Ltd |
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