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Value of staining intensity in the interpretation of immunohistochemistry for tumor markers in colorectal cancer

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Abstract

The purpose of this study was to determine whether staining intensity in conjunction with the percentage of positive tumor cells should be used as an indicator of protein expression detected by immunohistochemistry. A tissue microarray of 1,197 colorectal cancers was immunostained for p53, Her2/neu, epidermal growth factor receptor (EGFR), adenomatosis polyposis coli (APC), and β-catenin. Immunoreactivity was described by the percentage of positive tumor cells (percent positivity) and by the staining intensity (weak, moderate, strong). The interobserver reproducibility of both was evaluated by two pathologists. The association of T stage, N stage, tumor grade, vascular invasion, and survival with percent positivity, staining intensity, and the combination of both was assessed. In univariate analysis, protein expression assessed by percent positivity resulted in 11 significant associations between the proteins and clinico-pathological features. Eight of these 11 were also demonstrated using only the degree of staining intensity. However, more than half of the associations identified by percent positivity alone were lost when staining intensity was also analyzed in combination with the percentage of positive tumor cells. A scoring method based on percent positivity, rather than on staining intensity, for p53, Her2/neu, EGFR, APC, and β-catenin is reproducible and appears to be sufficient for establishing associations of the selected tumor markers with most clinico-pathological features.

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References

  1. Ansari-Lari M (2005) Letter to the editor: comparison between two test results, k statistic instead of simple overall agreement. Vet Parasitol 133:369–370

    Article  Google Scholar 

  2. Atkins D, Reiffen KA, Tegtmeier CL, Winther H, Bonato MS, Storkel S (2004) Immunohistochemical detection of EGFR in paraffin-embedded tumor tissues: variation in staining intensity due to choice of fixative and storage time of tissue sections. J Histochem Cytochem 52:893–901

    Article  PubMed  CAS  Google Scholar 

  3. Azria D, Bibeau F, Barbier N, Zouhair A, Lemanski C, Rouanet P, Ychou M, Senesse P, Ozsahin M, Pelegrin A, Dubois JB, Thezenas S (2005) Prognostic impact of epidermal growth factor receptor (EGFR) expression on loco-regional recurrence after preoperative radiotherapy in rectal cancer. BMC Cancer 5:62

    Article  PubMed  Google Scholar 

  4. Baker K, Zlobec I, Tornillo L, Terracciano L, Jass JR, Lugli A (2007) Differential significance of tumour infiltrating lymphocytes in sporadic mismatch repair deficient versus proficient colorectal cancers: a potential role for dysregulation of the transforming growth factor-beta pathway. Eur J Cancer 43:624–631

    Article  PubMed  CAS  Google Scholar 

  5. Barlund M, Forozan F, Kononen J, Bubendorf L, Chen Y, Bittner ML, Torhorst J, Haas P, Bucher C, Sauter G, Kallioniemi OP, Kallioniemi A (2000) Detecting activation of ribosomal protein S6 kinase by complementary DNA and tissue microarray analysis. J Natl Cancer Inst 92:1252–1259

    Article  PubMed  CAS  Google Scholar 

  6. Bibeau F, Boissiere-Michot F, Sabourin JC, Gourgou-Bourgade S, Radal M, Penault-Llorca F, Rochaix P, Arnould L, Bralet MP, Azria D, Ychou M (2006) Assessment of epidermal growth factor receptor (EGFR) expression in primary colorectal carcinomas and their related metastases on tissue sections and tissue microarray. Virchows Arch 449:281–287

    Article  PubMed  CAS  Google Scholar 

  7. Bravou V, Klironomos G, Papadaki E, Taraviras S, Varakis J (2006) ILK over- expression in human colon cancer progression correlates with activation of beta-catenin, down-regulation of E-cadherin and activation of the Akt-FKHR pathway. J Pathol 208:91–99

    Article  PubMed  CAS  Google Scholar 

  8. Garrity MM, Burgart LJ, Mahoney MR, Windschitl HE, Salim M, Wiesenfeld M, Krook JE, Michalak JC, Goldberg RM, O’Connell MJ, Furth AF, Sargent DJ, Murphy LM, Hill E, Riehle DL, Meyers CH, Witzig TE (2004) Prognostic value of proliferation, apoptosis, defective DNA mismatch repair, and p53 overexpression in patients with resected Dukes’ B2 or C colon cancer: a North Central Cancer Treatment Group Study. J Clin Oncol 22:1572–1582

    Article  PubMed  CAS  Google Scholar 

  9. Goethals L, Perneel C, Debucquoy A, De Schutter H, Borghys D, Ectors N, Geboes K, McBride WH, Haustermans KM (2006) A new approach to the validation of tissue microarrays. J Pathol 20:607–614

    Article  Google Scholar 

  10. Goldstein NS, Armin M (2001) Epidermal growth factor receptor immunohistochemical reactivity in patients with American Joint Committee on Cancer Stage IV colon adenocarcinoma: implications for a standardized scoring system. Cancer 92:1331–1346

    Article  PubMed  CAS  Google Scholar 

  11. Hampel H, Stephens JA, Pukkala E, Sankila R, Aaltonen LA, Mecklin JP, de la Chapelle A (2005) Cancer risk in hereditary nonpolyposis colorectal cancer syndrome: later age of onset. Gastroenterology 129:415–421

    Article  PubMed  Google Scholar 

  12. Italiano A, Saint-Paul MC, Caroli-Bosc FX, Francois E, Bourgeon A, Benchimol D, Gugenheim J, Michiels JF (2005) Epidermal growth factor receptor (EGFR) status in primary colorectal tumors correlates with EGFR expression in related metastatic sites: biological and clinical implications. Ann Oncol 16:1503–1507

    Article  PubMed  CAS  Google Scholar 

  13. Italiano A, Vandenbos FB, Otto J, Mouroux J, Fontaine D, Marcy PY, Cardot N, Thyss A, Pedeutour F (2006) Comparison of the epidermal growth factor receptor gene and protein in primary non-small-cell-lung cancer and metastatic sites: implications for treatment with EGFR-inhibitors. Ann Oncol 17:981–985

    Article  PubMed  CAS  Google Scholar 

  14. Kirkegaard T, Edwards J, Tovey S, McGlynn LM, Krishna SN, Mukherjee R, Tam L, Munro AF, Dunne B, Bartlett JM (2006) Observer variation in immunohistochemical analysis of protein expression, time for a change? Histopathology 48:787–794

    Article  PubMed  CAS  Google Scholar 

  15. Landis J, Koch G (1977) The measurement of observer agreement for categorical data. Biometrics 33:159–174

    Article  PubMed  CAS  Google Scholar 

  16. Lugli A, Zlobec I, Baker K, Minoo P, Tornillo L, Terracciano L, Jass J (2007) Prognostic significance of mucins in colorectal cancer with different DNA mismatch-repair status. J Clin Pathol 60:534–539

    Article  PubMed  CAS  Google Scholar 

  17. Lugli A, Zlobec I, Gunthert U, Minoo P, Baker K, Tornillo L, Terracciano L, Jass JR (2006) Overexpression of the receptor for hyaluronic acid mediated motility is an independent adverse prognostic factor in colorectal cancer. Mod Pathol 19:1302–1309

    Article  PubMed  CAS  Google Scholar 

  18. Lugli A, Zlobec I, Minoo P, Baker K, Tornillo L, Terracciano L, Jass JR (2006) Prognostic significance of the wnt signaling pathway molecules APC, β-catenin and E-cadherin in colorectal cancer. Histopathology 50:454–464

    Google Scholar 

  19. Lugli A, Zlobec I, Minoo P, Baker K, Tornillo L, Terracciano L, Jass JR (2006) Role of the mitogen-activated protein kinase and phosphoinositide 3-kinase/AKT pathways downstream molecules, phosphorylated extracellular signal-regulated kinase, and phosphorylated AKT in colorectal cancer-A tissue microarray-based approach. Human Pathol 37:1022–1031

    Article  CAS  Google Scholar 

  20. Minoo P, Zlobec I, Baker K, Tornillo L, Terracciano L, Jass JR, Lugli A (2006) Loss of Raf-1 kinase inhibitor protein (RKIP) expression is associated with tumor progression and metastasis in colorectal cancer. Am J Clin Pathol 127:820–827

    Article  Google Scholar 

  21. Minoo P, Zlobec I, Baker K, Tornillo L, Terracciano L, Jass JR, Lugli A (2007) Prognostic significance of mammalian sterile20-like kinase 1 in colorectal cancer. Mod Pathol 20:331–338

    Article  PubMed  CAS  Google Scholar 

  22. 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

    PubMed  CAS  Google Scholar 

  23. Resnick MB, Routhier J, Konkin T, Sabo E, Pricolo VE (2004) Epidermal growth factor receptor, c-MET, beta-catenin, and p53 expression as prognostic indicators in stage II colon cancer: a tissue microarray study. Clin Cancer Res 10:3069–3075

    Article  PubMed  CAS  Google Scholar 

  24. Sauter G, Simon R, Hillan K (2003) Tissue microarrays in drug discovery. Nat Rev Drug Discov 2:962–972

    Article  PubMed  CAS  Google Scholar 

  25. Schwandner O, Schiedeck TH, Bruch HP, Duchrow M, Windhoevel U (2000) p53 and Bcl-2 as significant predictors of recurrence and survival in rectal cancer. Eur J Cancer 36:348–356

    Article  PubMed  CAS  Google Scholar 

  26. Spano JP, Lagorce C, Atlan D, Milano G, Domont J, Benamouzig R, Attar A (2005) Impact of EGFR expression on colorectal cancer patient prognosis and survival. Ann Oncol 16:102–108

    Article  PubMed  Google Scholar 

  27. Takahashi Y, Bucana CD, Cleary KR, Ellis LM (1998) p53, vessel count, and vascular endothelial growth factor expression in human colon cancer. Int J Cancer 79:34–38

    Article  PubMed  CAS  Google Scholar 

  28. Torhorst J, Bucher C, Kononen J, Haas P, Zuber M, Kochli OR, Mross F (2001) Tissue microarrays for rapid linking of molecular changes to clinical endpoints. Am J Pathol 159:2249–2256

    PubMed  CAS  Google Scholar 

  29. Umemura S, Itoh J, Itoh H, Serizawa A, Saito Y, Suzuki Y, Tokuda Y, Tajima T, Osamura RY (2004) Immunohistochemical evaluation of hormone receptors in breast cancer: which scoring system is suitable for highly sensitive procedures? Appl Immunohistochem Mol Morphol 12:8–13

    PubMed  Google Scholar 

  30. Zlobec I, Steele R, Nigam N, Compton CC (2005) A predictive model of rectal tumor response to preoperative radiotherapy using classification and regression tree methods. Clin Cancer Res 11:5440–5443

    Article  PubMed  CAS  Google Scholar 

  31. Zlobec I, Steele R, Michel RP, Compton CC, Lugli A, Jass JR (2006) Scoring of p53, VEGF, Bcl-2 and APAF-1 immunohistochemistry and interobserver reliability in colorectal cancer. Mod Pathol 19:1236–1242

    Article  PubMed  CAS  Google Scholar 

  32. Zlobec I, Minoo P, Baker K, Haegert D, Khetani K, Tornillo L, Terracciano L, Jass JR, Lugli A (2007) Loss of APAF-1 expression is associated with tumour progression and adverse prognosis in colorectal cancer. Eur J Cancer 43:1101–1107

    Article  PubMed  CAS  Google Scholar 

  33. Zlobec I, Vuong T, Hayashi S, Haegert D, Tornillo L, Terracciano L, Lugli A, Jass J (2007) A simple and reproducible scoring system for EGFR in colorectal cancer: application to prognosis and prediction of response to preoperative brachytherapy. Br J Cancer 96:793–800

    Article  PubMed  CAS  Google Scholar 

  34. Zlobec I, Steele R, Terracciano L, Jass JR, Lugli A (2007) Selecting immunohisto-chemical cut-off scores for novel biomarkers of progression and survival in colorectal cancer. J Clin Pathol (in press). DOI 10.1136/jcp.2006.044537

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Authors and Affiliations

  1. Institute of Pathology, University Hospital of Basel, Schönbeinstrasse 40, Basel, 4031, Switzerland

    Inti Zlobec, Luigi Terracciano & Alessandro Lugli

  2. Department of Cellular Pathology, St. Mark’s Hospital, London, UK

    Jeremy R. Jass

  3. Imperial College, London, UK

    Jeremy R. Jass

Authors
  1. Inti Zlobec
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  2. Luigi Terracciano
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  3. Jeremy R. Jass
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  4. Alessandro Lugli
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Correspondence to Inti Zlobec.

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Zlobec, I., Terracciano, L., Jass, J.R. et al. Value of staining intensity in the interpretation of immunohistochemistry for tumor markers in colorectal cancer. Virchows Arch 451, 763–769 (2007). https://doi.org/10.1007/s00428-007-0466-8

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  • DOI: https://doi.org/10.1007/s00428-007-0466-8

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