Virchows Archiv

, Volume 444, Issue 2, pp 135–141

Microsatellite instability analysis and/or immunostaining for the diagnosis of hereditary nonpolyposis colorectal cancer?

  • Britta Halvarsson
  • Annika Lindblom
  • Eva Rambech
  • Kristina Lagerstedt
  • Mef Nilbert
Original Article

Abstract

Hereditary nonpolyposis colorectal cancer (HNPCC) represents 2–4% of colorectal cancers and is caused by a constitutional defect in a mismatch repair (MMR) gene, most commonly affecting the genes MLH1, MSH2, and MSH6. The MMR defect results in an increased cancer risk with the greatest lifetime risks for colorectal cancer and endometrial cancer. The HNPCC-associated tumor phenotype is generally characterized by microsatellite instability (MSI) and immunohistochemical loss of expression of the affected MMR protein. We have evaluated the information obtained from MSI analysis and immunostaining for MLH1, MSH2, and MSH6 in a series of 128 tumors from patients suspected of having HNPCC. A MSI-high pattern was present in 59 of 128 (46%) tumors. Loss of immunohistochemical expression for at least one of these MMR proteins was found in 54 of 59 (92%) evaluable MSI tumors. This loss affected MLH1 in 28, MSH2 in 22, and MSH6 in 21 tumors (with MSH6 as the only loss in 4 tumors). Five (8%) MSI-high tumors showed normal MMR protein expression. All 69 microsatellite stable or MSI-low tumors showed normal immunostaining for all three proteins. In 28 patients, all with MSI-H tumors, germ-line mutations of MLH1, MSH2, or MSH6 had been identified, and a corresponding immunohistochemical loss of MMR protein expression was identified in all these cases. In summary, immunostaining for the MMR proteins MLH1, MSH2, and MSH6 had a sensitivity of 92% and a specificity of 100% for detecting MMR-deficient tumors. MMR protein immunostaining facilitates mutation analysis in suspected HNPCC patients, since it pinpoints the mutated gene, but until the genetic background to the MSI tumors with retained MMR protein expression has been clarified, we suggest that MSI and MMR protein immunostaining should optimally be combined in clinical HNPCC analysis.

Keywords

Hereditary nonpolyposis colorectal cancer Microsatellite instability Immunohistochemistry 

References

  1. 1.
    Aaltonen LA, Salovaara R, Kristo P, Canzian F, Hemminki A, Peltomäki P, Chadwick RB, Kääriänen H, Eskelinen M, Järvinen H, Mecklin J-P, de la Chapelle A (1998) Incidence of hereditary nonpolyposis colorectal cancer and the feasibility of molecular screening for the disease. N Engl J Med 338:1481–1487PubMedGoogle Scholar
  2. 2.
    Aarnio M, Sankila R, Pukkala E, Salovaara R, Aaltonen LA, de la Chapelle A, Peltomäki P, Mecklin J-P, Järvinen HJ (1999) Cancer risk in mutation carriers of DNA mismatch-repair genes. Int J Cancer 81:214–218CrossRefPubMedGoogle Scholar
  3. 3.
    Boland CR, Thibodeau SN, Hamilton SR, Sidransky D, Eshleman JR, Burt RW, Meltzer SJ, Rodriguez-Bigas MA, Fodde R, Ranzani GN, Srivastava S (1998) 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 58:5248–5257PubMedGoogle Scholar
  4. 4.
    Chapusot C, Martin L, Bouvier AM, Bonithon-Kopp C, Ecarnot-Laubriet A, Rageot D, Ponnelle T, Laurent-Puig P, Faivre J, Piard F (2002) Microsatellite instability and intratumoural heterogeneity in 100 right-sided sporadic colon carcinomas. Br J Cancer 87:400–404CrossRefPubMedGoogle Scholar
  5. 5.
    Christensen M, Katballe M, Wikman F, Primdahl H, Sorensen FB, Laurberg S, Orntoft TF (2002) Antibody-based screening for hereditary nonpolyposis colorectal carcinoma compared with microsatellite instability analysis and sequencing. Cancer 95:2422–2430CrossRefPubMedGoogle Scholar
  6. 6.
    Debniak T, Kurzawski G, Gorski B, Kladny J, Domagaia W, Lubinski J (2000) Value of pedigree/clinical data, immunohistochemistry and microsatellite instability analyses in reducing the cost of determining hMLH1 and hMSH2 gene mutations in patients with colorectal cancer. Eur J Cancer 36:49–54CrossRefPubMedGoogle Scholar
  7. 7.
    Jass JR (2000) Familial colorectal cancer: pathology and molecular characteristics. Lancet Oncol 1:220–226CrossRefPubMedGoogle Scholar
  8. 8.
    Lanza G, Gafa R, Maestri I, Santini A, Matteuzzi M, Cavazzini L (2002) Immunohistochemical pattern of MLH1/MSH2 expression is related to clinical and pathological features in colorectal adenocarcinomas with microsatellite instability. Mod Pathol 15:741–749PubMedGoogle Scholar
  9. 9.
    Lindor NM, Burgart LJ, Leontovich O, Goldberg RM, Cunningham JM, Sargent DJ, Walsh-Vockley C, Petersen G, Walsh MD, Leggett BA, Young JP, Barker MA; Jass JR, Hopper J, Gallinger S, Bapat B, Redston M, Thibodeau SN (2002) Immunohistochemistry versus microsatellite instability testing in phenotyping colorectal tumors. J Clin Oncol 20:1043–1048PubMedGoogle Scholar
  10. 10.
    Mitchell RJ, Farrington SM, Dunlop MG, Campbell H (2002) Mismatch repair genes hMLH1 and hMSH2 in colorectal cancer: a huge review. Am J Epidemiol 156:885–902CrossRefPubMedGoogle Scholar
  11. 11.
    Moslein G, Tetsre DJ, Lindor NM, Honchel R, Cunningham JM, French AJ, Halling KC, Schwab M, Goretzki P, Thibodeau SN (1996) Microsatellite instability and mutation analysis of hMSH2 and hMLH1 in patients with sporadic, familial and hereditary colorectal cancer. Hum Mol Genet 5:1245–1252PubMedGoogle Scholar
  12. 12.
    Muc R and Naidoo R (2002) Microsatellite instability in diagnostic pathology. Curr Diagn Pathol 8:318–327CrossRefGoogle Scholar
  13. 13.
    Muller W, Burgart LJ, Krause-Paulus R, Thibodeau SN, Almeida M, Bocker Edmonston T, Boland CR, Sutter C, Jass JR, Lindblom A, Lubinsky J, Mac Dermott K, Sanders DSA, Morreau H, Muller A, Oliani C, Orntoft T, Ponz de Leon M, Rosty C, Rodriguez-Bigas M, Ruschoff J, Ruszkiewicz A, Sabourin J, Salovaara R, Möslein G and the ICG-HNPCC (2001) The reliability of Immunohistochemistry as a pre-screening method for the diagnosis of hereditary nonpolyposis colorectal cancer (HNPCC)—results of an international collaborative study. Familial Cancer 1:87–93CrossRefGoogle Scholar
  14. 14.
    Paraf F, Gilquin M, Longy M, Gilbert B, Gorry P, Petit B, Labrousse F (2001) MLH1 and MSH2 protein immunohistochemistry is useful for detection of hereditary non-polyposis colorectal cancer in young patients. Histopathology 39:250–258CrossRefGoogle Scholar
  15. 15.
    Peel DJ, Ziogas A, Fox EA, Gildea M, Laham B, Clements E, Kolodner RD, Anton-Cluver H (2000) Characterization of hereditary nonpolyposis colorectal cancer families from a population-based series of cases. J Natl Cancer Inst 92:1517–1522CrossRefPubMedGoogle Scholar
  16. 16.
    Plaschke J, Kruger S, Pistorius S, Theissig F, Saeger HD, Schackert HK (2002) Involvement of hMSH6 in the development of hereditary and sporadic colorectal cancer revealed by immunostaining is based on germline mutations, but rarely on somatic inactivation. Int J Cancer 97:643–648CrossRefPubMedGoogle Scholar
  17. 17.
    Rigau V, Sebbagh N, Olschwang S, Paraf F, Mourra N, Pacr Y, Flejou JF (2003) Microsatellite instability in colorectal carcinoma: the comparison of immunohistochemistry and molecular biology suggests a role for hMSH6 immunostaining. Arch Pathol Lab Med 127:694–700PubMedGoogle Scholar
  18. 18.
    Ruszkiewicz A, Bennett G, Moore J, Manavis J, Rudzki B, Shen L, Suthers G (2002) Correlation of mismatch repair genes immunohistochemistry and microsatellite instability status in HNPCC-associated tumours. Pathology 34:541–547PubMedGoogle Scholar
  19. 19.
    Vasen HF, van Ballegooijen M, Buskens E, Kleibeuker JK, Taal BG, Griffionen G, Nagengast FM, Menko FH, Meera Kahn P (1998) A cost-effectiveness analysis of colorectal screening of hereditary nonpolyposis colorectal carcinoma gene carriers. Cancer 82:1632–1697CrossRefPubMedGoogle Scholar
  20. 20.
    Vasen HF, Watson P, Mecklin J-P, Lynch HT and the ICG-HNPCC (1999) New clinical criteria for hereditary nonpolyposis colorectal cancer (HNPCC, Lynch syndrome) proposed by the International Collaborative Group on HNPCC. Gastroenterology 116:1453–1456PubMedGoogle Scholar
  21. 21.
    Ward R, Meagher A, Tomlinson I, O’Connor T, Norrie M, Wu R, Hawkins N (2001) Microsatellite instability and the clinicopathological features of sporadic colorectal cancer. Gut 48:821–829PubMedGoogle Scholar
  22. 22.
    Watson P, Lynch HT (2001) Cancer risk in mismatch repair gene mutation carriers. Fam Cancer 1:57–60CrossRefPubMedGoogle Scholar
  23. 23.
    Young J, Simms LA, Biden KG, Wynter C, Whitehall V, Karamatic R, George J, Goldblatt J, Walpole I, Robin S-A, Borten MM, Stitz R, Searle J, McKeone D, Fraser L, Purdie DR, Podger K, Price R, Buttenshaw R, Walsh MD, Barker M, Leggett BA, Jass JR (2001) Features of colorectal cancers with high-level microsatellite instability occurring in familial and sporadic setting. Am J Pathol 159:2107–2116PubMedGoogle Scholar
  24. 24.
    Yuen ST, Chan TL, Ho JWC, Chan ASY, Chung LP, Lam PWY, Wah Tse C, Wyllie AH, Leung SY (2002) Germline, somatic and epigenetic events underlying mismatch repair deficiency in colorectal and HNPCC-related cancers. Oncogene 21:7585–7592CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Britta Halvarsson
    • 1
    • 2
  • Annika Lindblom
    • 3
  • Eva Rambech
    • 4
  • Kristina Lagerstedt
    • 3
  • Mef Nilbert
    • 4
  1. 1.Department of Pathology, The Jubileum InstitutionLund University HospitalLundSweden
  2. 2.Department of PathologyHelsingborg HospitalHelsingborgSweden
  3. 3.Department of Clinical GeneticsKarolinska HospitalStockholmSweden
  4. 4.Department of OncologyThe Jubileum Institution, Lund University HospitalLundSweden

Personalised recommendations