NRAS Q61R , BRAF V600E immunohistochemistry: a concomitant tool for mutation screening in melanomas

Diagnostic Pathology

Pascale Marcorelles³  Cédric Le Marechal⁶  Marc De Braekeleer¹  Zarrin Alavi⁴  Laure Paule⁷  Laura Samaison⁵  Sebastian Costa⁸  Matthieu Talagas⁵  Arnaud Uguen² 

[1] CHRU Brest, Laboratoire de Cytogénétique et Biologie de la Reproduction, Brest F-29220, France;Université Européenne de Bretagne, Rennes, France;Department of Pathology, University Hospital Morvan, 2, Avenue Foch 29609, Brest, France;Inserm, CIC1412, Brest F-29200, France;Faculté de Médecine et des Sciences de la Santé Université de Brest, EA4685, Brest F-29200, France;Department of Molecular Genetics and Histocompatibily, University Hospital Morvan, 2, Avenue Foch 29609, Brest, France;CHRU Brest, Laboratoire de Génétique Moléculaire et d’Histocompatibilité, Brest F-29220, France;CHRU Brest, Service d’anatomie et cytologie pathologiques, Brest F-29220, France

关键词: Immunohistochemistry;    Molecular analysis;    NRAS;    BRAF;    Melanoma;   

Others  :  1225962
DOI  :  10.1186/s13000-015-0359-0

 received in 2015-04-03, accepted in 2015-07-09,  发布年份 2015

【 摘 要 】

Background

The determination of NRAS and BRAF mutation status is a major requirement in the treatment of patients with metastatic melanoma. Mutation specific antibodies against NRAS ^Q61Rand BRAF ^V600Eproteins could offer additional data on tumor heterogeneity. The specificity and sensitivity of NRAS ^Q61Rimmunohistochemistry have recently been reported excellent. We aimed to determine the utility of immunohistochemistry using SP174 anti-NRAS ^Q61Rand VE1 anti-BRAF ^V600Eantibodies in the theranostic mutation screening of melanomas.

Methods

142 formalin-fixed paraffin-embedded melanoma samples from 79 patients were analyzed using pyrosequencing and immunohistochemistry.

Results

23 and 26 patients were concluded to have a NRAS-mutated or a BRAF-mutated melanoma respectively. The 23 NRAS ^Q61R and 23 BRAF ^V600E -mutant samples with pyrosequencing were all positive in immunohistochemistry with SP174 antibody and VE1 antibody respectively, without any false negative. Proportions and intensities of staining were varied. Other NRAS ^Q61L , NRAS ^Q61K , BRAF ^V600K and BRAF ^V600R mutants were negative in immunohistochemistry. 6 single cases were immunostained but identified as wild-type using pyrosequencing (1 with SP174 and 5 with VE1). 4/38 patients with multiple samples presented molecular discordant data. Technical limitations are discussed to explain those discrepancies. Anyway we could not rule out real tumor heterogeneity.

Conclusions

In our study, we showed that combining immunohistochemistry analysis targeting NRAS ^Q61Rand BRAF ^V600Eproteins with molecular analysis was a reliable theranostic tool to face challenging samples of melanoma.

【 授权许可】

   
2015 Uguen et al.

【 图 表 】

Fig. 1.

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