Cellular Oncology

, Volume 35, Issue 5, pp 367–375 | Cite as

Molecular diagnosis of minimal residual disease in head and neck cancer patients

  • A. Peggy Graveland
  • Boudewijn J. M. Braakhuis
  • Simone E. J. Eerenstein
  • Remco de Bree
  • Elisabeth Bloemena
  • Michiel de Maaker
  • Michiel W. M. van den Brekel
  • Frederike Dijk
  • Wilma E. Mesker
  • Hans J. Tanke
  • C. Rene Leemans
  • Ruud H. Brakenhoff
Original Paper

Abstract

Aim

Locoregional recurrences and distant metastases in adequately treated head and neck squamous cell carcinoma (HNSCC) patients have a dismal effect on survival. Tumor cells that escape histopathological detection might be the prime cause of this effect. We evaluated whether minimal residual cancer (MRC) in deep surgical margins and disseminated tumor cells (DTCs) in bone marrow aspirates are associated with clinicohistopathological parameters and outcome.

Methods

Submucosal samples of deep resection margins of 105 HNSCC patients with histopathologically tumor-free surgical margins were analysed for the presence of MRC using hLy-6D qRT-PCR. Bone-marrow aspirates of 76 of these patients were analysed for DTCs by immunocytochemical staining. Presence of molecular-positive deep surgical margins, presence of DTC in bone marrow aspirates, and clinicohistopathological parameters were tested for associations with survival parameters by univariate and multivariate analyses.

Results

In addition to lymph node stage, it appeared that vasoinvasive growth and particularly infiltrative growth pattern are significant predictors for locoregional recurrence (p = 0.041 and p = 0.006, respectively) and disease–free survival (p = 0.014 and p = 0.008, respectively). Remarkably, neither the presence of molecular-positive deep surgical margins nor that of DTC in bone marrow aspirates were significantly related to outcome.

Conclusions

The presence of vasoinvasive and infiltrative growth in HNSCC tumor specimens are significant risk-factors for locoregional recurrence and disease-free survival. At present there seems no role for molecular analysis of deep surgical margins and bone marrow aspirates in predicting outcome with the methods used.

Keywords

Minimal residual cancer Bone marrow Head and neck cancer Non-cohesive growth Molecular diagnosis Vaso-invasive growth 

References

  1. 1.
    C.R. Leemans, R. Tiwari, J.J. Nauta, I. van der Waal, G.B. Snow, Recurrence at the primary site in head and neck cancer and the significance of neck lymph node metastases as a prognostic factor. Cancer 73, 187–190 (1994)PubMedCrossRefGoogle Scholar
  2. 2.
    H. Lumerman, P. Freedman, S. Kerpel, Oral epithelial dysplasia and the development of invasive squamous cell carcinoma. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 79, 321–329 (1995)PubMedCrossRefGoogle Scholar
  3. 3.
    S. Silverman Jr., M. Gorsky, G.E. Kaugars, Leukoplakia, dysplasia, and malignant transformation. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 82, 117 (1996)PubMedCrossRefGoogle Scholar
  4. 4.
    M. Weijers, G.B. Snow, D.P. Bezemer, J.E. van der Wal, I. van der Waal, The status of the deep surgical margins in tongue and floor of mouth squamous cell carcinoma and risk of local recurrence; an analysis of 68 patients. Int. J. Oral Maxillofac. Surg. 33, 146–149 (2004)PubMedCrossRefGoogle Scholar
  5. 5.
    M. Weijers, G.B. Snow, D.P. Bezemer, J.E. van der Wal, I. van der Waal, The clinical relevance of epithelial dysplasia in the surgical margins of tongue and floor of mouth squamous cell carcinoma: an analysis of 37 patients. J. Oral Pathol. Med. 31, 11–15 (2002)PubMedCrossRefGoogle Scholar
  6. 6.
    J.A. Woolgar, S. Rogers, C.R. West, R.D. Errington, J.S. Brown, E.D. Vaughan, Survival and patterns of recurrence in 200 oral cancer patients treated by radical surgery and neck dissection. Oral Oncol. 35, 257–265 (1999)PubMedCrossRefGoogle Scholar
  7. 7.
    J.A. Woolgar, J. Scott, E.D. Vaughan, J.S. Brown, C.R. West, S. Rogers, Survival, metastasis and recurrence of oral cancer in relation to pathological features. Ann. R. Coll. Surg. Engl. 77, 325–331 (1995)PubMedGoogle Scholar
  8. 8.
    F.A. Sawair, C.R. Irwin, D.J. Gordon, A.G. Leonard, M. Stephenson, S.S. Napier, Invasive front grading: reliability and usefulness in the management of oral squamous cell carcinoma. J. Oral Pathol. Med. 32, 1–9 (2003)PubMedCrossRefGoogle Scholar
  9. 9.
    V.M.M. van Houten, M.P. Tabor, M.W. van den Brekel, J.A. Kummer, F. Denkers, J. Dijkstra, C.R. Leemans, I. van der Waal, G.B. Snow, R.H. Brakenhoff, Mutated p53 as a molecular marker for the diagnosis of head and neck cancer. J. Pathol. 198, 476–486 (2002)PubMedCrossRefGoogle Scholar
  10. 10.
    B.J.M. Braakhuis, M.P. Tabor, J.A. Kummer, C.R. Leemans, R.H. Brakenhoff, A genetic explanation of Slaughter’s concept of field cancerization: evidence and clinical implications. Cancer Res. 63, 1727–1730 (2003)PubMedGoogle Scholar
  11. 11.
    M.P. Tabor, R.H. Brakenhoff, H.J. Ruijter-Schippers, J.A. Kummer, C.R. Leemans, B.J.M. Braakhuis, Genetically altered fields as origin of locally recurrent head and neck cancer: a retrospective study. Clin. Cancer Res. 10, 3607–3613 (2004)PubMedCrossRefGoogle Scholar
  12. 12.
    A.P. Graveland, P.J. Golusinski, M. Buijze, R. Douma, N. Sons, D.J. Kuik, E. Bloemena, C.R. Leemans, R.H. Brakenhoff, B.J.M. Braakhuis, Loss of heterozygosity at 9p and p53 immunopositivity in surgical margins predict local relapse in head and neck squamous cell carcinoma. Int. J. Cancer 128, 1852–1859 (2011)PubMedCrossRefGoogle Scholar
  13. 13.
    T.B. Schaaij-Visser, J.F. Bremmer, B.J.M. Braakhuis, A.J. Heck, M. Slijper, I. van der Waal, R.H. Brakenhoff, Evaluation of cornulin, keratin 4, keratin 13 expression and grade of dysplasia for predicting malignant progression of oral leukoplakia. Oral Oncol. 46, 123–127 (2010)PubMedCrossRefGoogle Scholar
  14. 14.
    J.A. Brennan, L. Mao, R.H. Hruban, J.O. Boyle, Y.J. Eby, W.M. Koch, S.N. Goodman, D. Sidransky, Molecular assessment of histopathological staging in squamous-cell carcinoma of the head and neck. N. Engl. J. Med. 332, 429–435 (1995)PubMedCrossRefGoogle Scholar
  15. 15.
    A.P. Graveland, M. de Maaker, B.J.M. Braakhuis, R. de Bree, S.E.J. Eerenstein, E. Bloemena, C.R. Leemans, R.H. Brakenhoff, Molecular detection of minimal residual cancer in surgical margins of head and neck cancer patients. Cell. Oncol. 31, 317–328 (2009)PubMedGoogle Scholar
  16. 16.
    W. Garavello, A. Ciardo, R. Spreafico, R.M. Gaini, Risk factors for distant metastases in head and neck squamous cell carcinoma. Arch. Otolaryngol. Head Neck Surg. 132, 762–766 (2006)PubMedCrossRefGoogle Scholar
  17. 17.
    O.R. Merino, R.D. Lindberg, G.H. Fletcher, An analysis of distant metastases from squamous cell carcinoma of the upper respiratory and digestive tracts. Cancer 40, 145–151 (1977)PubMedCrossRefGoogle Scholar
  18. 18.
    C. Kotwall, K. Sako, M.S. Razack, U. Rao, V. Bakamjian, D.P. Shed, Metastatic patterns in squamous cell cancer of the head and neck. Am. J. Surg. 154, 439–442 (1987)PubMedCrossRefGoogle Scholar
  19. 19.
    D.R. Colnot, E.J. Nieuwenhuis, D.J. Kuik, C.R. Leemans, J. Dijkstra, G.B. Snow, G.A.M.S. van Dongen, R.H. Brakenhoff, Clinical significance of micrometastatic cells detected by E48 (Ly-6D) reverse transcription-polymerase chain reaction in bone marrow of head and neck cancer patients. Clin. Cancer Res. 10, 7827–7833 (2004)PubMedCrossRefGoogle Scholar
  20. 20.
    S. Riethdorf, H. Wikman, K. Pantel, Review: biological relevance of disseminated tumor cells in cancer patients. Int. J. Cancer 123, 1991–2006 (2008)PubMedCrossRefGoogle Scholar
  21. 21.
    S. Braun, F.D. Vogl, B. Naume, W. Janni, M.P. Osborne, R.C. Coombes, G. Schlimok, I.J. Diel, B. Gerber, G. Gebauer, J.Y. Pierga, C. Marth, D. Oruzio, G. Wiedswang, E.F. Solomayer, G. Kundt, B. Strobl, T. Fehm, G.Y. Wong, J. Bliss, A. Vincent-Salomon, K. Pantel, A pooled analysis of bone marrow micrometastasis in breast cancer. N. Engl. J. Med. 353, 793–802 (2005)PubMedCrossRefGoogle Scholar
  22. 22.
    W. Janni, B. Rack, C. Schindlbeck, B. Strobl, D. Rjosk, S. Braun, H. Sommer, K. Pantel, B. Gerber, K. Friese, The persistence of isolated tumor cells in bone marrow from patients with breast carcinoma predicts an increased risk for recurrence. Cancer 103, 884–891 (2005)PubMedCrossRefGoogle Scholar
  23. 23.
    M. Partridge, R.H. Brakenhoff, E. Phillips, K. Ali, R. Francis, R. Hooper, K. Lavery, A. Brown, J. Langdon, Detection of rare disseminated tumor cells identifies head and neck cancer patients at risk of treatment failure. Clin. Cancer Res. 9, 5287–5294 (2003)PubMedGoogle Scholar
  24. 24.
    K. Shanmugaratnam, L.H. Sobin, The World Health Organization histological classification of tumours of the upper respiratory tract and ear. A commentary on the second edition. Cancer 71, 2689–2697 (1993)PubMedCrossRefGoogle Scholar
  25. 25.
    J.G. Batsakis, Surgical excision margins: a pathologist’s perspective. Adv. Anat. Pathol. 6, 140–148 (1999)PubMedCrossRefGoogle Scholar
  26. 26.
    T.R. Helliwell, J.A. Woolgar, Mucosal malignancies of the head and neck region. In Datasets for histopathology reports on head and neck carcinomas and salivary neoplasms, 2nd edn. The Royal College of Pathologists, London. http://www.rcpath.org (2005)
  27. 27.
    R.H. Brakenhoff, M. Gerretsen, E.M. Knippels, M. van Dijk, H. van Essen, D.O. Weghuis, R.J. Sinke, G.B. Snow, G.A.M.S. van Dongen, The human E48 antigen, highly homologous to the murine Ly-6 antigen ThB, is a GPI-anchored molecule apparently involved in keratinocyte cell-cell adhesion. J. Cell Biol. 129, 1677–1689 (1995)PubMedCrossRefGoogle Scholar
  28. 28.
    T.W. Geurts, P.M. Nederlof, M.W. van den Brekel, L.J. van’t Veer, D. de Jong, A.A. Hart, N. van Zandwijk, H. Klomp, A.J. Balm, M.L. van Velthuysen, Pulmonary squamous cell carcinoma following head and neck squamous cell carcinoma: metastasis or second primary? Clin. Cancer Res. 11, 6608–6614 (2005)PubMedCrossRefGoogle Scholar
  29. 29.
    S. Loseke, E. Grage-Griebenow, A. Wagner, K. Gehlhar, A. Bufe, Differential expression of IFN-alpha subtypes in human PBMC: evaluation of novel real-time PCR assays. J. Immunol. Methods 276, 207–222 (2003)PubMedCrossRefGoogle Scholar
  30. 30.
    E.J. Nieuwenhuis, C.R. Leemans, J.A. Kummer, F. Denkers, G.B. Snow, R.H. Brakenhoff, Assessment and clinical significance of micrometastases in lymph nodes of head and neck cancer patients detected by E48 (Ly-6D) quantitative reverse transcription-polymerase chain reaction. Lab. Investig. 83, 1233–1240 (2003)PubMedCrossRefGoogle Scholar
  31. 31.
    K. Pantel, G. Schlimok, M. Angstwurm, D. Weckermann, W. Schmaus, H. Gath, B. Passlick, J.R. Izbicki, G. Riethmuller, Methodological analysis of immunocytochemical screening for disseminated epithelial tumor cells in bone marrow. J. Hematother. 3, 165–173 (1994)PubMedCrossRefGoogle Scholar
  32. 32.
    F.S. Doekhie, W.E. Mesker, J.H. van Krieken, N.F. Kok, H.H. Hartgrink, E.K. Kranenbarg, H. Putter, P.J. Kuppen, H.J. Tanke, R.A. Tollenaar, C.J. van de Velde, Clinical relevance of occult tumor cells in lymph nodes from gastric cancer patients. Am. J. Surg. Pathol. 29, 1135–1144 (2005)PubMedCrossRefGoogle Scholar
  33. 33.
    T. Fehm, S. Braun, V. Muller, W. Janni, G. Gebauer, C. Marth, C. Schindlbeck, D. Wallwiener, E. Borgen, B. Naume, K. Pantel, E. Solomayer, A concept for the standardized detection of disseminated tumor cells in bone marrow from patients with primary breast cancer and its clinical implementation. Cancer 107, 885–892 (2006)PubMedCrossRefGoogle Scholar
  34. 34.
    K. Pantel, R.J. Cote, O. Fodstad, Detection and clinical importance of micrometastatic disease. J. Natl. Cancer Inst. 91, 1113–1124 (1999)PubMedCrossRefGoogle Scholar
  35. 35.
    M. Bryne, N. Jenssen, M. Boysen, Histological grading in the deep invasive front of T1 and T2 glottic squamous cell carcinomas has high prognostic value. Virchows Arch. 427, 277–281 (1995)PubMedCrossRefGoogle Scholar
  36. 36.
    W. Jerjes, T. Upile, A. Petrie, A. Riskalla, Z. Hamdoon, M. Vourvachis, K. Karavidas, A. Jay, A. Sandison, G.J. Thomas, N. Kalavrezos, C. Hopper, Clinicopathological parameters, recurrence, locoregional and distant metastasis in 115 T1-T2 oral squamous cell carcinoma patients. Head Neck Oncol. 2, 9 (2010)PubMedCrossRefGoogle Scholar

Copyright information

© International Society for Cellular Oncology 2012

Authors and Affiliations

  • A. Peggy Graveland
    • 1
  • Boudewijn J. M. Braakhuis
    • 1
  • Simone E. J. Eerenstein
    • 1
  • Remco de Bree
    • 1
  • Elisabeth Bloemena
    • 2
    • 3
  • Michiel de Maaker
    • 1
  • Michiel W. M. van den Brekel
    • 4
  • Frederike Dijk
    • 1
  • Wilma E. Mesker
    • 5
  • Hans J. Tanke
    • 6
  • C. Rene Leemans
    • 1
  • Ruud H. Brakenhoff
    • 1
  1. 1.Department of Otolaryngology/Head and Neck SurgeryVU University Medical CenterAmsterdamThe Netherlands
  2. 2.Department of PathologyVU University Medical CenterAmsterdamThe Netherlands
  3. 3.Department of Oral and Maxillofacial Surgery/Oral PathologyAcademic Centre of Dentistry Amsterdam (ACTA)AmsterdamThe Netherlands
  4. 4.Department of Head and Neck Oncology and SurgeryNetherlands Cancer Institute/Antoni van Leeuwenhoek Hospital and Institute of Phonetic Sciences University of Amsterdam (ACLC)AmsterdamThe Netherlands
  5. 5.Department of SurgeryLeiden University Medical CenterLeidenThe Netherlands
  6. 6.Department of Molecular Cell BiologyLeiden University Medical CenterLeidenThe Netherlands

Personalised recommendations