Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma

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dc.contributor.authorRosa, Fabiola E. [UNESP]
dc.contributor.authorSilveira, Sara M. [UNESP]
dc.contributor.authorSilveira, Cassia G. T. [UNESP]
dc.contributor.authorBergamo, Nadia A. [UNESP]
dc.contributor.authorMoraes Neto, Francisco A.
dc.contributor.authorDomingues, Maria Aparecida Custódio [UNESP]
dc.contributor.authorSoares, Fernando A.
dc.contributor.authorCaldeira, Jose R. F. [UNESP]
dc.contributor.authorRogatto, Silvia Regina [UNESP]
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionAmaral Carvalho Hosp
dc.contributor.institutionHosp AC Camargo Fund Antonio Prudente
dc.contributor.institutionHosp AC Camargo Liberdade São Paulo
dc.date.accessioned2014-05-20T13:37:22Z
dc.date.available2014-05-20T13:37:22Z
dc.date.issued2009-03-23
dc.description.abstractBackground: 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.en
dc.description.affiliationUNESP São Paulo State Univ, Fac Med, Dept Urol, Botucatu, SP, Brazil
dc.description.affiliationUNESP São Paulo State Univ, Inst Biosci, Dept Genet, Botucatu, SP, Brazil
dc.description.affiliationAmaral Carvalho Hosp, Dept Pathol, São Paulo, Brazil
dc.description.affiliationUNESP São Paulo State Univ, Fac Med, Dept Pathol, Botucatu, SP, Brazil
dc.description.affiliationHosp AC Camargo Fund Antonio Prudente, Fundação Antonio Prudente, Dept Pathol, São Paulo, Brazil
dc.description.affiliationAmaral Carvalho Hosp, Dept Senol, São Paulo, Brazil
dc.description.affiliationHosp AC Camargo Liberdade São Paulo, Fundação Antonio Prudente 211, NeoGene Lab, São Paulo, Brazil
dc.description.affiliationUnespUNESP São Paulo State Univ, Fac Med, Dept Urol, Botucatu, SP, Brazil
dc.description.affiliationUnespUNESP São Paulo State Univ, Inst Biosci, Dept Genet, Botucatu, SP, Brazil
dc.description.affiliationUnespUNESP São Paulo State Univ, Fac Med, Dept Pathol, Botucatu, SP, Brazil
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.format.extent12
dc.identifierhttp://dx.doi.org/10.1186/1471-2407-9-90
dc.identifier.citationBmc Cancer. London: Biomed Central Ltd., v. 9, p. 12, 2009.
dc.identifier.doi10.1186/1471-2407-9-90
dc.identifier.fileWOS000265611700001.pdf
dc.identifier.issn1471-2407
dc.identifier.lattes2259986546265579
dc.identifier.urihttp://hdl.handle.net/11449/12920
dc.identifier.wosWOS:000265611700001
dc.language.isoeng
dc.publisherBiomed Central Ltd.
dc.relation.ispartofBMC Cancer
dc.relation.ispartofjcr3.288
dc.relation.ispartofsjr1,464
dc.rights.accessRightsAcesso aberto
dc.sourceWeb of Science
dc.titleQuantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinomaen
dc.typeArtigo
dcterms.licensehttp://www.biomedcentral.com/about/license
dcterms.rightsHolderBiomed Central Ltd.
unesp.author.lattes2259986546265579
unesp.author.orcid0000-0003-1647-7842[7]
unesp.author.orcid0000-0002-0932-7769[3]
unesp.campusUniversidade Estadual Paulista (Unesp), Instituto de Biociências, Botucatupt
unesp.campusUniversidade Estadual Paulista (Unesp), Faculdade de Medicina, Botucatupt

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