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Immunohistochemical staining for p16 and BRAFV600E is useful to distinguish between sporadic and hereditary (Lynch syndrome-related) microsatellite instable colorectal carcinomas

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Abstract

DNA mismatch repair (MMR) protein analysis by immunohistochemistry (IHC) can identify colorectal cancer (CRC) with microsatellite instability (MSI). As MLH1-deficient CRC can be hereditary or sporadic, markers to distinguish between them are needed. MLH1 promoter methylation assay is the reference method; however, sometimes, it is challenging on formalin-fixed paraffin-embedded tissue samples. We assessed by IHC the expression of BRAFV600E, p16, MGMT, and CDX2 in 55 MLH1-deficient MSI CRC samples (of which 8 had a germline MLH1 mutation) to determine whether this panel differentiates between sporadic and hereditary CRCs. We also analyzed MLH1 promoter methylation by methylation-specific PCR and pyrosequencing and BRAF status by genotyping. None of the hereditary CRCs showed MLH1 methylation, BRAF mutation, BRAFV600E-positive immunostaining, or loss of p16 expression. We detected MLH1 promoter methylation in 67 % and a BRAF mutation in 42 % of CRC, all showing MLH1 promoter methylation. BRAFV600E IHC and BRAF genotyping gave concordant results in all but two samples. Loss of expression of p16 was found in 30 % of CRC with methylation of the MLH1 promoter, but its expression was retained in all non-methylated and part of MLH1-methylated tumors (100 % specificity, 30 % sensitivity). CDX2 and MGMT expression was not associated with MLH1 status. Thus, BRAFV600E and p16 IHC may help in differentiating sporadic from hereditary MLH1-deficient CRC with MSI. Specifically, p16 IHC might be used as a surrogate marker for MLH1 promoter methylation, because all p16-negative CRCs displayed MLH1 methylation, whereas hereditary CRCs were all p16-positive.

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Acknowledgments

We thank Dr. Hélène de Forges for her help in editing the manuscript.

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Authors and Affiliations

  1. Pathology Department, Institut Régional du Cancer de Montpellier, 208 rue des Apothicaires, 34 298, Montpellier Cedex 5, France

    Florence Boissière-Michot, Hélène Frugier & Frédéric Bibeau

  2. Translational Research Department, Institut Régional du Cancer de Montpellier, 208 rue des Apothicaires, 34 298, Montpellier Cedex 5, France

    Florence Boissière-Michot, Alexandre Ho-Pun-Cheung & Evelyne Lopez-Crapez

  3. Oncogenetic Department, Institut Régional du Cancer de Montpellier, 208 rue des Apothicaires, 34 298, Montpellier Cedex 5, France

    Jacqueline Duffour

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  1. Florence Boissière-Michot
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Correspondence to Florence Boissière-Michot.

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Boissière-Michot, F., Frugier, H., Ho-Pun-Cheung, A. et al. Immunohistochemical staining for p16 and BRAFV600E is useful to distinguish between sporadic and hereditary (Lynch syndrome-related) microsatellite instable colorectal carcinomas. Virchows Arch 469, 135–144 (2016). https://doi.org/10.1007/s00428-016-1958-1

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