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Protocolo de estudio molecular del oncogénHER2/neu en el carcinoma de mama

Molecular protocol forHER2/neu analysis in breast carcinoma

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Resumen

Antecedentes

El oncogénHER2/neu es un determinante biológico predictivo esencial en el cáncer de mama por su papel como diana terapéutica del trastuzumab. No obstante, no se ha llegado aún a consensuar un método de referencia para su estudio.

Métodos

En este trabajo, hemos aplicado y comparado, sobre 222 carcinomas de mama recopilados prospectivamente, los dos ensayos de uso más frecuente para determinar el estado deHER2/neu: inmunohistoquímica para expresión de la proteína p185 empleando dos anticuerpos monoclonales (CB11 y TAB250) e hibridaciónin situ fluorescente (FISH), utilizando secciones de tejido incluido en parafina.

Resultados

Los resultados demuestran mayor sensibilidad y especificidad con CB11 (62,5% y 93,4% respectivamente) que con TAB250 (40% y 76,4% respectivamente). El estudio inmunohistoquímico con CB11 es adecuado como prueba inicial para predecir el estado del gen, proporciona un alto valor predictivo negativo (95,5%) en los casos de expresión débil o nula y positivo (96,2%) en los tumores con sobreexpresión. Si la expresión inmunohistoquímica es moderada debe considerarse no concluyente y es indispensable el estudio de FISH. También es recomendable aplicar FISH si hay discordancia entre la expresión de p185 y el perfil morfológico y/o molecular tumoral. Finalmente, aunque la tasa de falsos positivos producidos por estudios inmunohistoquímicos es inferior al 5%, debido a la toxicidad y coste de la terapia con trastuzumab es razonable considerar el empleo sistemático de FISH antes de indicar el tratamiento.

Conclusión

En base a nuestros resultados proponemos un protocolo para el estudio molecular deHER2/neu.

Abstract

Background

The HER2/neu proto-oncogene is frequently over-expressed in breast cancer and serves as a biological target for trastuzumab therapy. However, there is no consensus regarding the technical aspects to be used to define HER2/neu status in clinical practice.

Methods

The present study was conducted to address this critical issue by prospectively analysing a large cohort of breast cancer patients (n=222) and using a variety of methods. To define HER2/neu expression, detection of its encoded protein (p185) was performed by comparative immunohistochemical (IHC) analysis using two mouse monoclonal antibodies (mAb CB11 and mAb TAB250). To assess HER2/neu gene amplification, fluorescentin situ hybridisation (FISH) assays with gene-specific probes were conducted. All procedures were applied to de-paraffinised tissue sections of breast tumour samples.

Results

Results showed that mAb CB11 had increased sensitivity and specificity (62.5% and 93.4%, respectively) compared to mAb TAB250 (40% and 76.4%, respectively) in defining HER2/neu amplification. We conclude that HER2/neu measurement by IHC using mAb CB11 is an appropriate strategy which provides a high negative predictive value (95.5%) for HER2/neu amplification in cases with low or undetectable p185 expression. Conversely, mAb CB11 has a high positive predictive value (96.2%) for HER2/neu amplification in cases with p185 overexpression. However, cases with moderate p185 expression need to be considered as inconclusive. In such cases, it is necessary to use FISH measurement to evaluate HER2/neu amplification. It is also advisable to conduct FISH if there is discordance between p185 expression and the histopathology classification of the lesion, or molecular profile of the tumour. Finally, even though the false positive rate of IHC assay is <5%, the toxicity and cost of trastuzumab therapy suggest that FISH be used systematically prior to implementation of treatment.

Conclusion

We suggest the use of a molecular protocol for HER2/neu analysis in this type of tumor.

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Authors and Affiliations

  1. HISTOPAT Laboratoris, Barcelona, España

    Montse Verdú Artufel, Abelardo Moreno Carazo, Carlos Cordón Cardó & Xavier Puig Torrus

  2. BIOPAT. Biopatología Molecular, Barcelona, España

    Anna Colomer Valero, Ruth Román Lladó, Nadina Erill Sagalés, Carlos Cordón Cardó & Xavier Puig Torrus

  3. Departmento de Estadística de la Universidad de Barcelona, Barcelona, España

    Miquel Calvo Llorca

  4. Division of Molecular Pathology, Memorial Sloan-Kettering Cancer Center, New York, USA

    Carlos Cordón Cardó

Authors
  1. Montse Verdú Artufel
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  2. Anna Colomer Valero
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  3. Ruth Román Lladó
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  4. Nadina Erill Sagalés
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  5. Miquel Calvo Llorca
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  6. Abelardo Moreno Carazo
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  7. Carlos Cordón Cardó
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  8. Xavier Puig Torrus
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Correspondence to Xavier Puig Torrus.

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Artufel, M.V., Valero, A.C., Lladó, R.R. et al. Protocolo de estudio molecular del oncogénHER2/neu en el carcinoma de mama. Clin Transl Oncol 7, 504–511 (2005). https://doi.org/10.1007/BF02717004

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  • DOI: https://doi.org/10.1007/BF02717004

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