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. 2004 Sep;41(9):664–668. doi: 10.1136/jmg.2004.020651

BRAF screening as a low-cost effective strategy for simplifying HNPCC genetic testing

E Domingo 1, P Laiho 1, M Ollikainen 1, M Pinto 1, L Wang 1, A French 1, J Westra 1, T Frebourg 1, E Espin 1, M Armengol 1, R Hamelin 1, H Yamamoto 1, R Hofstra 1, R Seruca 1, A Lindblom 1, P Peltomaki 1, S Thibodeau 1, L Aaltonen 1, S Schwartz 1
PMCID: PMC1735885  PMID: 15342696

Abstract

Background: According to the international criteria for hereditary non-polyposis colorectal cancer (HNPCC) diagnostics, cancer patients with a family history or early onset of colorectal tumours showing high microsatellite instability (MSI-H) should receive genetic counselling and be offered testing for germline mutations in DNA repair genes, mainly MLH1 and MSH2. Recently, an oncogenic V600E hotspot mutation within BRAF, a kinase encoding gene from the RAS/RAF/MAPK pathway, has been found to be associated with sporadic MSI-H colon cancer, but its association with HNPCC remains to be further clarified.

Methods: BRAF-V600E mutations were analysed by automatic sequencing in colorectal cancers from 206 sporadic cases with MSI-H and 111 HNPCC cases with known germline mutations in MLH1 and MSH2. In addition, 45 HNPCC cases showing abnormal immunostaining for MSH2 were also analysed.

Results: The BRAF-V600E hotspot mutation was found in 40% (82/206) of the sporadic MSI-H tumours analysed but in none of the 111 tested HNPCC tumours or in the 45 cases showing abnormal MSH2 immunostaining.

Conclusions: Detection of the V600E mutation in a colorectal MSI-H tumour argues against the presence of a germline mutation in either the MLH1 or MSH2 gene. Therefore, screening of these mismatch repair (MMR) genes can be avoided in cases positive for V600E if no other significant evidence, such as fulfilment of the strict Amsterdam criteria, suggests MMR associated HNPCC. In this context, mutation analysis of the BRAF hotspot is a reliable, fast, and low cost strategy which simplifies genetic testing for HNPCC.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aaltonen L. A., Peltomäki P., Leach F. S., Sistonen P., Pylkkänen L., Mecklin J. P., Järvinen H., Powell S. M., Jen J., Hamilton S. R. Clues to the pathogenesis of familial colorectal cancer. Science. 1993 May 7;260(5109):812–816. doi: 10.1126/science.8484121. [DOI] [PubMed] [Google Scholar]
  2. Aaltonen L. A., Salovaara R., Kristo P., Canzian F., Hemminki A., Peltomäki P., Chadwick R. B., Käriäinen H., Eskelinen M., Järvinen H. Incidence of hereditary nonpolyposis colorectal cancer and the feasibility of molecular screening for the disease. N Engl J Med. 1998 May 21;338(21):1481–1487. doi: 10.1056/NEJM199805213382101. [DOI] [PubMed] [Google Scholar]
  3. Boland C. R., Thibodeau S. N., Hamilton S. R., Sidransky D., Eshleman J. R., Burt R. W., Meltzer S. J., Rodriguez-Bigas M. A., Fodde R., Ranzani G. N. A National Cancer Institute Workshop on Microsatellite Instability for cancer detection and familial predisposition: development of international criteria for the determination of microsatellite instability in colorectal cancer. Cancer Res. 1998 Nov 15;58(22):5248–5257. [PubMed] [Google Scholar]
  4. Cunningham J. M., Kim C. Y., Christensen E. R., Tester D. J., Parc Y., Burgart L. J., Halling K. C., McDonnell S. K., Schaid D. J., Walsh Vockley C. The frequency of hereditary defective mismatch repair in a prospective series of unselected colorectal carcinomas. Am J Hum Genet. 2001 Aug 24;69(4):780–790. doi: 10.1086/323658. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davies Helen, Bignell Graham R., Cox Charles, Stephens Philip, Edkins Sarah, Clegg Sheila, Teague Jon, Woffendin Hayley, Garnett Mathew J., Bottomley William. Mutations of the BRAF gene in human cancer. Nature. 2002 Jun 9;417(6892):949–954. doi: 10.1038/nature00766. [DOI] [PubMed] [Google Scholar]
  6. Deng Guoren, Bell Ian, Crawley Suzanne, Gum James, Terdiman Jonathan P., Allen Brian A., Truta Brindusa, Sleisenger Marvin H., Kim Young S. BRAF mutation is frequently present in sporadic colorectal cancer with methylated hMLH1, but not in hereditary nonpolyposis colorectal cancer. Clin Cancer Res. 2004 Jan 1;10(1 Pt 1):191–195. doi: 10.1158/1078-0432.ccr-1118-3. [DOI] [PubMed] [Google Scholar]
  7. Di Fiore F., Charbonnier F., Martin C., Frerot S., Olschwang S., Wang Q., Boisson C., Buisine M-P, Nilbert M., Lindblom A. Screening for genomic rearrangements of the MMR genes must be included in the routine diagnosis of HNPCC. J Med Genet. 2004 Jan;41(1):18–20. doi: 10.1136/jmg.2003.012062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Domingo Enric, Espín Eloi, Armengol Manel, Oliveira Carla, Pinto Mafalda, Duval Alex, Brennetot Caroline, Seruca Raquel, Hamelin Richard, Yamamoto Hiroyuki. Activated BRAF targets proximal colon tumors with mismatch repair deficiency and MLH1 inactivation. Genes Chromosomes Cancer. 2004 Feb;39(2):138–142. doi: 10.1002/gcc.10310. [DOI] [PubMed] [Google Scholar]
  9. Fallik David, Borrini Francesco, Boige Valérie, Viguier Jérôme, Jacob Sandrine, Miquel Catherine, Sabourin Jean-Christophe, Ducreux Michel, Praz Françoise. Microsatellite instability is a predictive factor of the tumor response to irinotecan in patients with advanced colorectal cancer. Cancer Res. 2003 Sep 15;63(18):5738–5744. [PubMed] [Google Scholar]
  10. Gille J. J. P., Hogervorst F. B. L., Pals G., Wijnen J. Th, van Schooten R. J., Dommering C. J., Meijer G. A., Craanen M. E., Nederlof P. M., de Jong D. Genomic deletions of MSH2 and MLH1 in colorectal cancer families detected by a novel mutation detection approach. Br J Cancer. 2002 Oct 7;87(8):892–897. doi: 10.1038/sj.bjc.6600565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hoang J. M., Cottu P. H., Thuille B., Salmon R. J., Thomas G., Hamelin R. BAT-26, an indicator of the replication error phenotype in colorectal cancers and cell lines. Cancer Res. 1997 Jan 15;57(2):300–303. [PubMed] [Google Scholar]
  12. Ionov Y., Peinado M. A., Malkhosyan S., Shibata D., Perucho M. Ubiquitous somatic mutations in simple repeated sequences reveal a new mechanism for colonic carcinogenesis. Nature. 1993 Jun 10;363(6429):558–561. doi: 10.1038/363558a0. [DOI] [PubMed] [Google Scholar]
  13. Kane M. F., Loda M., Gaida G. M., Lipman J., Mishra R., Goldman H., Jessup J. M., Kolodner R. Methylation of the hMLH1 promoter correlates with lack of expression of hMLH1 in sporadic colon tumors and mismatch repair-defective human tumor cell lines. Cancer Res. 1997 Mar 1;57(5):808–811. [PubMed] [Google Scholar]
  14. Kumar Rajiv, Angelini Sabrina, Hemminki Kari. Activating BRAF and N-Ras mutations in sporadic primary melanomas: an inverse association with allelic loss on chromosome 9. Oncogene. 2003 Dec 18;22(58):9217–9224. doi: 10.1038/sj.onc.1206909. [DOI] [PubMed] [Google Scholar]
  15. Lynch H. T., de la Chapelle A. Genetic susceptibility to non-polyposis colorectal cancer. J Med Genet. 1999 Nov;36(11):801–818. [PMC free article] [PubMed] [Google Scholar]
  16. Marra G., Boland C. R. Hereditary nonpolyposis colorectal cancer: the syndrome, the genes, and historical perspectives. J Natl Cancer Inst. 1995 Aug 2;87(15):1114–1125. doi: 10.1093/jnci/87.15.1114. [DOI] [PubMed] [Google Scholar]
  17. Miyaki M., Konishi M., Tanaka K., Kikuchi-Yanoshita R., Muraoka M., Yasuno M., Igari T., Koike M., Chiba M., Mori T. Germline mutation of MSH6 as the cause of hereditary nonpolyposis colorectal cancer. Nat Genet. 1997 Nov;17(3):271–272. doi: 10.1038/ng1197-271. [DOI] [PubMed] [Google Scholar]
  18. Nyström-Lahti M., Kristo P., Nicolaides N. C., Chang S. Y., Aaltonen L. A., Moisio A. L., Järvinen H. J., Mecklin J. P., Kinzler K. W., Vogelstein B. Founding mutations and Alu-mediated recombination in hereditary colon cancer. Nat Med. 1995 Nov;1(11):1203–1206. doi: 10.1038/nm1195-1203. [DOI] [PubMed] [Google Scholar]
  19. Peltomäki P., Lothe R. A., Aaltonen L. A., Pylkkänen L., Nyström-Lahti M., Seruca R., David L., Holm R., Ryberg D., Haugen A. Microsatellite instability is associated with tumors that characterize the hereditary non-polyposis colorectal carcinoma syndrome. Cancer Res. 1993 Dec 15;53(24):5853–5855. [PubMed] [Google Scholar]
  20. Rajagopalan Harith, Bardelli Alberto, Lengauer Christoph, Kinzler Kenneth W., Vogelstein Bert, Velculescu Victor E. Tumorigenesis: RAF/RAS oncogenes and mismatch-repair status. Nature. 2002 Aug 29;418(6901):934–934. doi: 10.1038/418934a. [DOI] [PubMed] [Google Scholar]
  21. Ramsey S. D., Clarke L., Etzioni R., Higashi M., Berry K., Urban N. Cost-effectiveness of microsatellite instability screening as a method for detecting hereditary nonpolyposis colorectal cancer. Ann Intern Med. 2001 Oct 16;135(8 Pt 1):577–588. doi: 10.7326/0003-4819-135-8_part_1-200110160-00008. [DOI] [PubMed] [Google Scholar]
  22. Reyes Carolina M., Allen Brian A., Terdiman Jonathan P., Wilson Leslie S. Comparison of selection strategies for genetic testing of patients with hereditary nonpolyposis colorectal carcinoma: effectiveness and cost-effectiveness. Cancer. 2002 Nov 1;95(9):1848–1856. doi: 10.1002/cncr.10910. [DOI] [PubMed] [Google Scholar]
  23. Ribic Christine M., Sargent Daniel J., Moore Malcolm J., Thibodeau Stephen N., French Amy J., Goldberg Richard M., Hamilton Stanley R., Laurent-Puig Pierre, Gryfe Robert, Shepherd Lois E. Tumor microsatellite-instability status as a predictor of benefit from fluorouracil-based adjuvant chemotherapy for colon cancer. N Engl J Med. 2003 Jul 17;349(3):247–257. doi: 10.1056/NEJMoa022289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rodriguez-Bigas M. A., Boland C. R., Hamilton S. R., Henson D. E., Jass J. R., Khan P. M., Lynch H., Perucho M., Smyrk T., Sobin L. A National Cancer Institute Workshop on Hereditary Nonpolyposis Colorectal Cancer Syndrome: meeting highlights and Bethesda guidelines. J Natl Cancer Inst. 1997 Dec 3;89(23):1758–1762. doi: 10.1093/jnci/89.23.1758. [DOI] [PubMed] [Google Scholar]
  25. Salovaara R., Loukola A., Kristo P., Käriäinen H., Ahtola H., Eskelinen M., Härkönen N., Julkunen R., Kangas E., Ojala S. Population-based molecular detection of hereditary nonpolyposis colorectal cancer. J Clin Oncol. 2000 Jun;18(11):2193–2200. doi: 10.1200/JCO.2000.18.11.2193. [DOI] [PubMed] [Google Scholar]
  26. Thibodeau S. N., Bren G., Schaid D. Microsatellite instability in cancer of the proximal colon. Science. 1993 May 7;260(5109):816–819. doi: 10.1126/science.8484122. [DOI] [PubMed] [Google Scholar]
  27. Vasen H. F., Mecklin J. P., Khan P. M., Lynch H. T. The International Collaborative Group on Hereditary Non-Polyposis Colorectal Cancer (ICG-HNPCC). Dis Colon Rectum. 1991 May;34(5):424–425. doi: 10.1007/BF02053699. [DOI] [PubMed] [Google Scholar]
  28. Vasen H. F., Watson P., Mecklin J. P., Lynch H. T. New clinical criteria for hereditary nonpolyposis colorectal cancer (HNPCC, Lynch syndrome) proposed by the International Collaborative group on HNPCC. Gastroenterology. 1999 Jun;116(6):1453–1456. doi: 10.1016/s0016-5085(99)70510-x. [DOI] [PubMed] [Google Scholar]
  29. Veigl M. L., Kasturi L., Olechnowicz J., Ma A. H., Lutterbaugh J. D., Periyasamy S., Li G. M., Drummond J., Modrich P. L., Sedwick W. D. Biallelic inactivation of hMLH1 by epigenetic gene silencing, a novel mechanism causing human MSI cancers. Proc Natl Acad Sci U S A. 1998 Jul 21;95(15):8698–8702. doi: 10.1073/pnas.95.15.8698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wahlberg S., Liu T., Lindblom P., Lindblom A. Various mutation screening techniques in the DNA mismatch repair genes hMSH2 and hMLH1. Genet Test. 1999;3(3):259–264. doi: 10.1089/109065799316563. [DOI] [PubMed] [Google Scholar]
  31. Wang Liang, Cunningham Julie M., Winters Jennifer L., Guenther Jennifer C., French Amy J., Boardman Lisa A., Burgart Lawrence J., McDonnell Shannon K., Schaid Daniel J., Thibodeau Stephen N. BRAF mutations in colon cancer are not likely attributable to defective DNA mismatch repair. Cancer Res. 2003 Sep 1;63(17):5209–5212. [PubMed] [Google Scholar]

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