Virchows Archiv

, Volume 463, Issue 5, pp 613–621 | Cite as

BRAF mutation in sporadic colorectal cancer and Lynch syndrome

  • Alexandra Thiel
  • Mira Heinonen
  • Jonas Kantonen
  • Annette Gylling
  • Laura Lahtinen
  • Mari Korhonen
  • Soili Kytölä
  • Jukka-Pekka Mecklin
  • Arto Orpana
  • Päivi Peltomäki
  • Ari RistimäkiEmail author
Original Article


The aim of the study was to detect mutations of BRAF oncogene in colorectal cancer and to use this information to identify Lynch syndrome patients. Consecutive cases of primary colorectal cancer (n = 137) were analyzed for MLH1 protein expression using immunohistochemistry (IHC). BRAF V600E mutation was detected by IHC using a specific monoclonal antibody (VE1) and by qPCR. All MLH1 protein-negative cases were subjected to microsatellite instability analysis and MLH1 promoter methylation assay. MLH1 protein expression deficiency and high microsatellite instability (MSI-H) were detected in 18 of the 137 (13.1 %) consecutive colorectal cancer specimens. Detection of the BRAF V600E mutation by IHC was 100 % sensitive and specific as compared to qPCR, and this mutation was frequently present in the MSI-H group (77.8 %; 14/18) and less frequently in the microsatellite-stable group (7.6 %; 9/118). All BRAF V600E mutated cases of the MSI-H group presented with a MLH1 promoter methylation (14/14) as detected by methylation-specific multiplex ligation-dependent probe amplification. When BRAF was wild type in the MSI-H group, only one MLH1 promoter methylation was detected (1/4), and of the remaining three cases without MLH1 methylation, two were identified to harbor an MLH1 mutation consistent with Lynch syndrome. Finally, 11 previously confirmed Lynch syndrome cases were analyzed for BRAF V600E mutation, and all of them were wild type. In conclusion, detection of BRAF V600E in colorectal cancer specimens by IHC is sensitive and specific and may help to identify Lynch syndrome patients.


BRAF Colorectal cancer Lynch syndrome MLH1 Microsatellite instability Methylation 



We thank Carita Liikanen, Erkki Hänninen, and Helena Ahola for excellent technical assistance, and Mia Kero and Satu Remes for technical advice. We thank Dr. Pirjo Nummela for comments on the manuscript. This work was supported in part by grants from the Academy of Finland (grant no. 257795, PP), Sigrid Juselius Foundation (J-PM, PP, AR), Finnish Cancer Organization (J-PM, PP, AR), Biocentrum Helsinki (PP), FP7-ERC-232635 (PP), and Helsinki University Central Hospital Research Funds and HUSLAB R&D Funds (AR).

Conflict of interest

We declare that we have no conflict of interest.

Supplementary material

428_2013_1470_Fig3_ESM.jpg (151 kb)
Supplemental Fig. 1

Immunohistochemical staining of melanoma and colorectal cancer specimens with the commercial BRAF V600E mutation specific monoclonal antibody (dilution 1:2,000) with different detection systems. a Melanoma specimen displaying strong BRAF V600E staining. b Colorectal cancer (CRC) specimen with relatively strong BRAF V600E staining. c Colorectal cancer specimen with moderate BRAF V600E staining intensity. Cancer glands with BRAF V600E mutation are stained whereas normal glands (right side of the picture) remain unstained. For all specimens (a-c) the staining intensity decreases dramatically when no amplification is applied and with the UltraView detection system (with or without amplification). AMP (amplification), CRC (colorectal cancer). Original magnification is 100x (JPEG 151 kb)

428_2013_1470_MOESM1_ESM.tif (11.9 mb)
High Resolution Image (TIFF 12,180 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Alexandra Thiel
    • 1
    • 2
  • Mira Heinonen
    • 1
    • 2
  • Jonas Kantonen
    • 1
    • 2
  • Annette Gylling
    • 3
  • Laura Lahtinen
    • 4
  • Mari Korhonen
    • 5
  • Soili Kytölä
    • 5
  • Jukka-Pekka Mecklin
    • 6
  • Arto Orpana
    • 5
  • Päivi Peltomäki
    • 3
  • Ari Ristimäki
    • 1
    • 2
    Email author
  1. 1.Division of Pathology and Genetics, HUSLAB, and Haartman InstituteHelsinki University Central Hospital and University of HelsinkiHelsinkiFinland
  2. 2.Genome-Scale Biology, Research Programs UnitUniversity of HelsinkiHelsinkiFinland
  3. 3.Department of Medical GeneticsUniversity of HelsinkiHelsinkiFinland
  4. 4.Department of PathologyCentral Hospital Central FinlandJyväskyläFinland
  5. 5.Laboratory of GeneticsHelsinki University Central Hospital, HUSLABHelsinkiFinland
  6. 6.Department of Surgery, Jyväskylä Central Hospital and Institute of Clinical MedicineUniversity of Eastern FinlandJyväskyläFinland

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