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Familial Cancer

, Volume 6, Issue 3, pp 301–310 | Cite as

Incorporation of somatic BRAF mutation testing into an algorithm for the investigation of hereditary non-polyposis colorectal cancer

  • M. B. LoughreyEmail author
  • P. M. Waring
  • A. Tan
  • M. Trivett
  • S. Kovalenko
  • V. Beshay
  • M.-A. Young
  • G. McArthur
  • A. Boussioutas
  • A. Dobrovic
Article

Abstract

Patients suspected on clinical grounds to have hereditary non-polyposis colorectal cancer (HNPCC) may be offered laboratory testing in order to confirm the diagnosis and to facilitate screening of pre-symptomatic family members. Tumours from an affected family member are usually pre-screened for microsatellite instability (MSI) and/or loss of immunohistochemical expression of mismatch repair (MMR) genes prior to germline MMR gene mutation testing. The efficiency of this triage process is compromised by the more frequent occurrence of sporadic colorectal cancer (CRC) showing high levels of MSI (MSI-H) due to epigenetic loss of MLH1 expression. Somatic BRAF mutations, most frequently V600E, have been described in a significant proportion of sporadic MSI-H CRC but not in HNPCC-associated cancers. BRAF mutation testing has therefore been proposed as a means to more definitively identify and exclude sporadic MSI-H CRC cases from germline MMR gene testing. However, the clinical validity and utility of this approach have not been previously evaluated in a familial cancer clinic setting. Testing for the V600E mutation was performed on MSI-H CRC samples from 68 individuals referred for laboratory investigation of suspected HNPCC. The V600E mutation was identified in 17 of 40 (42%) tumours showing loss of MLH1 protein expression by immunohistochemistry but in none of the 28 tumours that exhibited loss of MSH2 expression (P < 0.001). The assay was negative in all patients with an identified germline MMR gene mutation. Although biased by the fact that germline testing was not pursued beyond direct sequencing in many cases lacking a high clinical index of suspicion of HNPCC, BRAF V600E detection was therefore considered to be 100% specific and 48% sensitive in detecting sporadic MSI-H CRC amongst those cases showing loss of MLH1 protein expression, in a population of patients with MSI-H CRC and clinical features suggestive of HNPCC. Accordingly, we recommend the incorporation of BRAF V600E mutation testing into the laboratory algorithm for pre-screening patients with suspected HNPCC, whose CRCs show loss of expression of MLH1. In such tumours, the presence of a BRAF V600E mutation indicates the tumour is not related to HNPCC and that germline testing of MLH1 in that individual is not warranted. We also recommend that in families where the clinical suspicion of HNPCC is high, germline testing should not be performed on an individual whose CRC harbours a somatic BRAF mutation, as this may compromise identification of the familial mutation.

Keywords

Hereditary non-polyposis colorectal cancer BRAF Allele-specific PCR Microsatellite instability 

Abbreviations

HNPCC

Hereditary non-polyposis colorectal cancer

MSI

Microsatellite instability

MMR

Mismatch repair

CRC

Colorectal cancer

AS-PCR

Allele-specific polymerase chain reaction

Notes

Acknowledgements

M. B. Loughrey was supported by a Cancer Council of Victoria scholarship. We are grateful to Dr. Desiree du Sart of the Molecular Genetics Laboratory, Victorian Clinical Genetics Service for the provision of the results of MLPA analysis.

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Copyright information

© Springer Science + Business Media B.V. 2007

Authors and Affiliations

  • M. B. Loughrey
    • 1
    • 2
    Email author
  • P. M. Waring
    • 3
    • 4
  • A. Tan
    • 1
  • M. Trivett
    • 3
  • S. Kovalenko
    • 3
  • V. Beshay
    • 3
  • M.-A. Young
    • 5
  • G. McArthur
    • 6
  • A. Boussioutas
    • 7
    • 8
  • A. Dobrovic
    • 1
    • 9
  1. 1.Molecular Pathology Research LaboratoryPeter MacCallum Cancer CentreMelbourneAustralia
  2. 2.Department of PathologyRoyal Group of HospitalsBelfastN. Ireland, UK
  3. 3.Department of PathologyPeter MacCallum Cancer CentreMelbourneAustralia
  4. 4.Genentech Inc.South San FranciscoUSA
  5. 5.Familial Cancer CentrePeter MacCallum Cancer CentreMelbourneAustralia
  6. 6.Division of Haematology and Medical OncologyPeter MacCallum Cancer CentreMelbourneAustralia
  7. 7.Trescowthick Research LaboratoryPeter MacCallum Cancer CentreMelbourneAustralia
  8. 8.Department of MedicineUniversity of MelbourneMelbourneAustralia
  9. 9.Department of PathologyUniversity of MelbourneMelbourneAustralia

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