Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma
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 UNESP |
Texto Completo: | http://dx.doi.org/10.1186/1471-2407-9-90 http://hdl.handle.net/11449/12920 |
Resumo: | Background: HER-2 gene testing has become an integral part of breast cancer patient diagnosis. The most commonly used assay in the clinical setting for evaluating HER-2 status is immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). These procedures permit correlation between HER-2 expression and morphological features. However, FISH signals are labile and fade over time, making post-revision of the tumor difficult. CISH (chromogenic in situ hybridization) is an alternative procedure, with certain advantages, although still limited as a diagnostic tool in breast carcinomas.Methods: To elucidate the molecular profile of HER-2 status, mRNA and protein expression in 75 invasive breast carcinomas were analyzed by real time quantitative RT-PCR (qRT-PCR) and IHC, respectively. Amplifications were evaluated in 43 of these cases by CISH and in 11 by FISH.Results: The concordance rate between IHC and qRT-PCR results was 78.9%, and 94.6% for qRT-PCR and CISH. Intratumoral heterogeneity of HER-2 status was identified in three cases by CISH. The results of the three procedures were compared and showed a concordance rate of 83.8%; higher discordances were observed in 0 or 1+immunostaining cases, which showed high-level amplification (15.4%) and HER-2 transcript overexpression (20%). Moreover, 2+ immunostaining cases presented nonamplified status (50%) by CISH and HER-2 downexpression (38.5%) by qRT-PCR. In general, concordance occurred between qRT-PCR and CISH results. A high concordance was observed between CISH/qRT-PCR and FISH. Comparisons with clinicopathological data revealed a significant association between HER-2 downexpression and the involvement of less than four lymph nodes (P = 0.0350).Conclusion: Based on these findings, qRT-PCR was more precise and reproducible than IHC. Furthermore, CISH was revealed as an alternative and useful procedure for investigating amplifications involving the HER-2 gene. |
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Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinomaBackground: HER-2 gene testing has become an integral part of breast cancer patient diagnosis. The most commonly used assay in the clinical setting for evaluating HER-2 status is immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). These procedures permit correlation between HER-2 expression and morphological features. However, FISH signals are labile and fade over time, making post-revision of the tumor difficult. CISH (chromogenic in situ hybridization) is an alternative procedure, with certain advantages, although still limited as a diagnostic tool in breast carcinomas.Methods: To elucidate the molecular profile of HER-2 status, mRNA and protein expression in 75 invasive breast carcinomas were analyzed by real time quantitative RT-PCR (qRT-PCR) and IHC, respectively. Amplifications were evaluated in 43 of these cases by CISH and in 11 by FISH.Results: The concordance rate between IHC and qRT-PCR results was 78.9%, and 94.6% for qRT-PCR and CISH. Intratumoral heterogeneity of HER-2 status was identified in three cases by CISH. The results of the three procedures were compared and showed a concordance rate of 83.8%; higher discordances were observed in 0 or 1+immunostaining cases, which showed high-level amplification (15.4%) and HER-2 transcript overexpression (20%). Moreover, 2+ immunostaining cases presented nonamplified status (50%) by CISH and HER-2 downexpression (38.5%) by qRT-PCR. In general, concordance occurred between qRT-PCR and CISH results. A high concordance was observed between CISH/qRT-PCR and FISH. Comparisons with clinicopathological data revealed a significant association between HER-2 downexpression and the involvement of less than four lymph nodes (P = 0.0350).Conclusion: Based on these findings, qRT-PCR was more precise and reproducible than IHC. Furthermore, CISH was revealed as an alternative and useful procedure for investigating amplifications involving the HER-2 gene.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)UNESP São Paulo State Univ, Fac Med, Dept Urol, Botucatu, SP, BrazilUNESP São Paulo State Univ, Inst Biosci, Dept Genet, Botucatu, SP, BrazilAmaral Carvalho Hosp, Dept Pathol, São Paulo, BrazilUNESP São Paulo State Univ, Fac Med, Dept Pathol, Botucatu, SP, BrazilHosp AC Camargo Fund Antonio Prudente, Fundação Antonio Prudente, Dept Pathol, São Paulo, BrazilAmaral Carvalho Hosp, Dept Senol, São Paulo, BrazilHosp AC Camargo Liberdade São Paulo, Fundação Antonio Prudente 211, NeoGene Lab, São Paulo, BrazilUNESP São Paulo State Univ, Fac Med, Dept Urol, Botucatu, SP, BrazilUNESP São Paulo State Univ, Inst Biosci, Dept Genet, Botucatu, SP, BrazilUNESP São Paulo State Univ, Fac Med, Dept Pathol, Botucatu, SP, BrazilBiomed Central Ltd.Universidade Estadual Paulista (Unesp)Amaral Carvalho HospHosp AC Camargo Fund Antonio PrudenteHosp AC Camargo Liberdade São PauloRosa, Fabiola E. [UNESP]Silveira, Sara M. [UNESP]Silveira, Cassia G. T. [UNESP]Bergamo, Nadia A. [UNESP]Moraes Neto, Francisco A.Domingues, Maria Aparecida Custódio [UNESP]Soares, Fernando A.Caldeira, Jose R. F. [UNESP]Rogatto, Silvia Regina [UNESP]2014-05-20T13:37:22Z2014-05-20T13:37:22Z2009-03-23info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article12application/pdfhttp://dx.doi.org/10.1186/1471-2407-9-90Bmc Cancer. London: Biomed Central Ltd., v. 9, p. 12, 2009.1471-2407http://hdl.handle.net/11449/1292010.1186/1471-2407-9-90WOS:000265611700001WOS000265611700001.pdf2259986546265579Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBMC Cancer3.2881,464info:eu-repo/semantics/openAccess2024-09-03T14:29:59Zoai:repositorio.unesp.br:11449/12920Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestrepositoriounesp@unesp.bropendoar:29462024-09-03T14:29:59Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma |
title |
Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma |
spellingShingle |
Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma Rosa, Fabiola E. [UNESP] |
title_short |
Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma |
title_full |
Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma |
title_fullStr |
Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma |
title_full_unstemmed |
Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma |
title_sort |
Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma |
author |
Rosa, Fabiola E. [UNESP] |
author_facet |
Rosa, Fabiola E. [UNESP] Silveira, Sara M. [UNESP] Silveira, Cassia G. T. [UNESP] Bergamo, Nadia A. [UNESP] Moraes Neto, Francisco A. Domingues, Maria Aparecida Custódio [UNESP] Soares, Fernando A. Caldeira, Jose R. F. [UNESP] Rogatto, Silvia Regina [UNESP] |
author_role |
author |
author2 |
Silveira, Sara M. [UNESP] Silveira, Cassia G. T. [UNESP] Bergamo, Nadia A. [UNESP] Moraes Neto, Francisco A. Domingues, Maria Aparecida Custódio [UNESP] Soares, Fernando A. Caldeira, Jose R. F. [UNESP] Rogatto, Silvia Regina [UNESP] |
author2_role |
author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Amaral Carvalho Hosp Hosp AC Camargo Fund Antonio Prudente Hosp AC Camargo Liberdade São Paulo |
dc.contributor.author.fl_str_mv |
Rosa, Fabiola E. [UNESP] Silveira, Sara M. [UNESP] Silveira, Cassia G. T. [UNESP] Bergamo, Nadia A. [UNESP] Moraes Neto, Francisco A. Domingues, Maria Aparecida Custódio [UNESP] Soares, Fernando A. Caldeira, Jose R. F. [UNESP] Rogatto, Silvia Regina [UNESP] |
description |
Background: HER-2 gene testing has become an integral part of breast cancer patient diagnosis. The most commonly used assay in the clinical setting for evaluating HER-2 status is immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). These procedures permit correlation between HER-2 expression and morphological features. However, FISH signals are labile and fade over time, making post-revision of the tumor difficult. CISH (chromogenic in situ hybridization) is an alternative procedure, with certain advantages, although still limited as a diagnostic tool in breast carcinomas.Methods: To elucidate the molecular profile of HER-2 status, mRNA and protein expression in 75 invasive breast carcinomas were analyzed by real time quantitative RT-PCR (qRT-PCR) and IHC, respectively. Amplifications were evaluated in 43 of these cases by CISH and in 11 by FISH.Results: The concordance rate between IHC and qRT-PCR results was 78.9%, and 94.6% for qRT-PCR and CISH. Intratumoral heterogeneity of HER-2 status was identified in three cases by CISH. The results of the three procedures were compared and showed a concordance rate of 83.8%; higher discordances were observed in 0 or 1+immunostaining cases, which showed high-level amplification (15.4%) and HER-2 transcript overexpression (20%). Moreover, 2+ immunostaining cases presented nonamplified status (50%) by CISH and HER-2 downexpression (38.5%) by qRT-PCR. In general, concordance occurred between qRT-PCR and CISH results. A high concordance was observed between CISH/qRT-PCR and FISH. Comparisons with clinicopathological data revealed a significant association between HER-2 downexpression and the involvement of less than four lymph nodes (P = 0.0350).Conclusion: Based on these findings, qRT-PCR was more precise and reproducible than IHC. Furthermore, CISH was revealed as an alternative and useful procedure for investigating amplifications involving the HER-2 gene. |
publishDate |
2009 |
dc.date.none.fl_str_mv |
2009-03-23 2014-05-20T13:37:22Z 2014-05-20T13:37:22Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1186/1471-2407-9-90 Bmc Cancer. London: Biomed Central Ltd., v. 9, p. 12, 2009. 1471-2407 http://hdl.handle.net/11449/12920 10.1186/1471-2407-9-90 WOS:000265611700001 WOS000265611700001.pdf 2259986546265579 |
url |
http://dx.doi.org/10.1186/1471-2407-9-90 http://hdl.handle.net/11449/12920 |
identifier_str_mv |
Bmc Cancer. London: Biomed Central Ltd., v. 9, p. 12, 2009. 1471-2407 10.1186/1471-2407-9-90 WOS:000265611700001 WOS000265611700001.pdf 2259986546265579 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
BMC Cancer 3.288 1,464 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
12 application/pdf |
dc.publisher.none.fl_str_mv |
Biomed Central Ltd. |
publisher.none.fl_str_mv |
Biomed Central Ltd. |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
repositoriounesp@unesp.br |
_version_ |
1810021377188560896 |