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Tissue microarray technology: validation in colorectal carcinoma and analysis of p53, hMLH1, and hMSH2 immunohistochemical expression

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Abstract

Tissue microarray technology enables the analysis of hundreds of specimens by arranging numerous 0.6-mm tissue core biopsy specimens into a single paraffin block. Validation studies are necessary to evaluate the representativeness of small disks taken from the original tissue. We validated the tissue microarray technology in colorectal carcinoma by analyzing the immunohistochemical expression of proteins involved in the two main pathways of colorectal carcinogenesis: p53 protein for loss of heterozygosity tumors, hMLH1 and hMSH2 proteins for microsatellite instability (MSI) tumors. We compared in 30 colorectal carcinomas (15 MSI− and 15 MSI+), 8 microarrays disks, and the whole section of the block from which they were derived. Tumoral tissue was present in 95.7% of the microarray disks. The analysis of three disks per case was comparable to the analysis of the whole section in 99.6% (p53), 98.8% (hMLH1), and 99.2% (hMSH2) of cases. In the second part we applied the tissue microarray technology to 263 consecutive cases of colorectal carcinoma, sampled by three cores. We showed that 48.5% overexpressed p53 and 8.7% lost hMLH1 or hMSH2. Tissue microarray technology, validated in colorectal carcinoma, appears as a useful research tool for rapid analysis of the clinical interest of molecular alterations.

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References

  1. Allegra CJ, Parr AL, Wold LE, Mahoney MR, Sargent DJ, Johnston P, Klein P, Behan K, O'Connell MJ, Levitt R, Kugler JW, Tirona MT, Goldberg RM (2002) Investigation of the prognostic and predictive value of thymidylate synthase, p53, and Ki-67 in patients with locally advanced colon cancer. J Clin Oncol 20:1735–1743

    Article  CAS  PubMed  Google Scholar 

  2. Battifora H (1986) Methods in laboratory investigation. Lab Invest 55:244–248

    CAS  PubMed  Google Scholar 

  3. Bocker Edmonston T, Cuesta KH, Burkholder S, Barusevicius A, Rose D, Kovatich AJ, Boman B, Fry R, Fishel R, Palazzo JP (2000) Colorectal carcinomas with high microsatellite instability: defining a distinct immunologic and molecular entity with respect to prognostic markers. Hum Pathol 31:1506–1514

    CAS  PubMed  Google Scholar 

  4. Boland RC, 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 58:5248–5257

    PubMed  Google Scholar 

  5. Bubendorf L, Nocito A, Moch H, Sauter G (2001) Tissue microarray (TMA) technology: miniaturized pathology archives for high-throughput in situ studies. J Pathol 195:72–79

    Article  CAS  PubMed  Google Scholar 

  6. Camp RL, Charette LA, Rimm DL (2000) Validation of tissue microarray technology in breast carcinoma. Lab Invest 80:1943–1949

    CAS  PubMed  Google Scholar 

  7. Chaves P, Cruz C, Lage P, Claro I, Cravo M, Leitão CN, Soares J (2000) Immunohistochemical detection of mismatch repair gene proteins as a useful tool for identification of colorectal carcinoma with the mutator phenotype. J Pathol 191:355–360

    Article  CAS  PubMed  Google Scholar 

  8. Chung DC (2000) The genetic basis of colorectal cancer: insights into critical pathways of tumorigenesis. Gastroenterology 119:854–865

    CAS  PubMed  Google Scholar 

  9. Elsaleh H, Joseph D, Grieu F, Zeps N, Spry N, Iacopetta B (2000) Association of tumour site and sex with survival benefit from adjuvant chemotherapy in colorectal cancer. Lancet 355:1745–1750

    PubMed  Google Scholar 

  10. Fernebro E, Dictor M, Bendahl PO, Fernö M, Nilbert M (2002) Evaluation of the Tissue Microarray technique for immunohistochemical analysis in rectal cancer. Arch Pathol Lab Med 126:702–705

    PubMed  Google Scholar 

  11. Fukushima H, Yamamoto H, Itoh F, Horiuchi S, Min Y, Iku S, Imai K (2001) Frequent alterations of the β-catenin and TCF-4 genes, but not of the APC gene, in colon cancers with high-frequency microsatellite instability. J Exp Clin Cancer Res 20:533–559

    Google Scholar 

  12. Gafà R, Maestri I, Matteuzzi M, Santini A, Ferretti S, Gavazzini L, Lanza G (2000) Sporadic colorectal adenocarcinoma with high-frequency microsatellite instability: pathobiologic features, hMLH1 and hMSH2 expression and clinical outcome. Cancer 89:2025–2037

    Article  CAS  PubMed  Google Scholar 

  13. Gardner MJ, Altman DG (2000) Confidence interval and statistical guidelines. In: Statistics with confidence, 2nd edn. British Medical Journal, London, pp 116–118

  14. Giardiello FM, Brensinger JD, Petersen GM (2001) AGA technical review on hereditary colorectal cancer and genetic testing. Gastroenterology 121:198–213

    CAS  PubMed  Google Scholar 

  15. Gillett CE, Springall RJ, Barnes DM, Hanby AM (2000) Multiple tissue core arrays in histopathology research: a validation study. J Pathol 192:549–553

    Article  CAS  PubMed  Google Scholar 

  16. Gryfe R, Kim H, Hsieh E TK, Aronson MD, Holowaty EJ, Bull SB, Redston M, Gallinger S (2000) Tumor microsatellite instability and clinical outcome in young patients with colorectal cancer. N Engl J Med 342:69–77

    CAS  PubMed  Google Scholar 

  17. Hamelin R, Laurent-Puig P (1997) p53 et cancers colorectaux. Pathol Biol (Paris) 45:876–881

    Google Scholar 

  18. Hao X, Frayling I, Willcocks T, Han W, Tomlinson I, Pignatelli M, Pretlow T, Talbot I (2002) β-Catenin expression and allelic loss at APC in sporadic colorectal carcinogenesis. Virchows Arch 440:362–366

    Article  CAS  PubMed  Google Scholar 

  19. Hoos A, Urist MJ, Stojadinovic A, Mastorides S, Dudas ME, Leung DH, Kuo D, Brennan MF, Lewis JJ, Cordon-Cardo C (2001) Validation of tissue microarrays for immunohistochemical profiling of cancer specimens using the example of human fibroblastic tumors. Am J Pathol 158:1245–1251

    CAS  PubMed  Google Scholar 

  20. Jiricny J, Nyström-Lahti M (2000) Mismatch repair defects in cancer. Curr Opin Genet Dev 10:157–161

    CAS  PubMed  Google Scholar 

  21. Kononen J, Bubendorf L, Kallioniemi A, Bärlund M, Schraml P, Leighton S, Torhorst J, Mihatsch MJ, Sauter G, Kallioniemi OP (1998) Tissue microarrays for high-throughput molecular profiling of tumor specimens. Nat Med 4:844–847

    CAS  PubMed  Google Scholar 

  22. Laurent-Puig P, Blons H, Cugnenc P-H (1999) Sequence of molecular genetic events in colorectal tumorigenesis. Eur J Cancer Prev 8:S39–S47

    PubMed  Google Scholar 

  23. Leahy DT, Salman R, Mulcahy H, Sheahan K, O'Donoghue DP, Parfrey NA (1996) Prognostic significance of p53 abnormalities in colorectal carcinoma detected by PCR-SSCP and immunohistochemical analysis. J Pathol 180:364–370

    Article  CAS  PubMed  Google Scholar 

  24. Lindor NM, Burgart LJ, Leontovich O, Goldberg RM, Cunningham JM, Sargent DJ, Walsh-Vockley C, Petersen GM, 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 20:1043–1048

    Google Scholar 

  25. Manne U, Myers RB, Moron C, Poczatek RB, Dillard D, Weiss H, Brown D, Srivastava S, Grizzle WE (1997) Prognostic significance of Bcl-2 expression and p53 nuclear accumulation in colorectal adenocarcinoma. Int J Cancer 74:346–358

    Article  CAS  PubMed  Google Scholar 

  26. Miyaki M, Iijima T, Kimura J, Yasuno M, Mori T, Hayashi Y, Koike M, Shitara N, Iwama T, Kuroki T (1999) Frequent mutation of β-catenin and APC genes in primary colorectal tumors from patients with hereditary nonpolyposis colorectal cancer. Cancer Res 59:4506–4509

    CAS  PubMed  Google Scholar 

  27. Moch H, Schraml P, Bubendorf L, Mirlacher M, Kononen J, Gasser T, Mihatsch MJ, Kallioniemi OP, Sauter G (1999) High-throughput tissue microarray analysis to evaluate genes uncovered by cDNA microarray screening in renal cell carcinoma. Am J Pathol 154:981–986

    CAS  PubMed  Google Scholar 

  28. Mucci NR, Akdas G, Manely S, Rubin MA (2000) Neuroendocrine expression in metastatic prostate cancer: evaluation of high throughput tissue microarrays to detect heterogeneous protein expression. Hum Pathol 31:406–414

    CAS  PubMed  Google Scholar 

  29. Nocito A, Bubendorf L, Tinner EM, Süess K, Wagner U, Forster T, Kononen J, Fijan A, Bruderer J, Schmid U, Ackermann D, Maurer R, Alund G, Knönagel H, Rist M, Anabitarte M, Hering F, Hardmeier T, Schoenenberger AJ, Flury R, Jäger P, Fehr JL, Schraml P, Moch H, Mihatsch MJ, Gasser T, Sauter G (2001) Microarrays of bladder cancer tissue are highly representative of proliferation index and histological grade. J Pathol 194:349–357

    CAS  PubMed  Google Scholar 

  30. Otsuka M, Kato M, Yoshikawa T, Chen H, Brown EJ, Masuho Y, Omata M, Seki N (2001) Differential expression of the L-plastin gene in human colorectal cancer progression and metastasis. Biochem Biophys Res Commun 289:876–881

    Article  CAS  PubMed  Google Scholar 

  31. Rigau V, Sebbagh N, Olschwang S, Paraf F, Mourra N, Parc Y, Fléjou JF (2003) Microsatellite instability in colorectal carcinoma: the comparison of immunohistochemistry and molecular biology suggests the interest of hMLH6 immunostaining. Arch Pathol Lab Med 127:694–700

    Google Scholar 

  32. Samowitz WS, Holden JA, Curtin K, Edwards SL, Walker AR, Lin HA, Robertson MA, Nichols MF, Gruenthal KM, Lynch BJ, Leppert MF, Slattery ML (2001) Inverse relationship between microsatellite instability and K-ras and p53 gene alterations in colon cancer. Am J Pathol 158:1517–1524

    CAS  PubMed  Google Scholar 

  33. Torhorst J, Bucher C, Kononen J, Hass P, Zuber M, Köchli O, Mross F, Dieterich H, Moch H, Mihatsch M, Kallioniemi OP, Sauter G (2001) Tissue microarrays for rapid linking of molecular changes to clinical endpoints. Am J Pathol 159:2249–2256

    PubMed  Google Scholar 

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Acknowledgements

F.J. was supported by a grant from the Association pour la Recherche sur le Cancer. We acknowledge the Association Charles Debray for its financial support.

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Authors and Affiliations

  1. Department of Pathology, Hôpital Saint-Antoine, AP-HP, 184 rue du Faubourg Saint-Antoine, 75012, Paris, France

    Florence Jourdan, Nicole Sebbagh, Eva Comperat, Najat Mourra & Jean-François Flejou

  2. Unité INSERM 434, 10 rue Juliette Dodu, 75010, Paris, France

    Florence Jourdan, Nicole Sebbagh, Sylviane Olschwang, Alex Duval, Richard Hamelin & Jean-François Flejou

  3. Unité INSERM 444, Faculté de Médecine Saint-Antoine, 27 rue Chaligny, 75571, Paris cedex 12, France

    Antoine Flahault

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  1. Florence Jourdan
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Correspondence to Jean-François Flejou.

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Jourdan, F., Sebbagh, N., Comperat, E. et al. Tissue microarray technology: validation in colorectal carcinoma and analysis of p53, hMLH1, and hMSH2 immunohistochemical expression. Virchows Arch 443, 115–121 (2003). https://doi.org/10.1007/s00428-003-0833-z

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  • DOI: https://doi.org/10.1007/s00428-003-0833-z

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