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British Journal of Cancer logoLink to British Journal of Cancer
. 2000 Aug 16;83(5):674–683. doi: 10.1054/bjoc.2000.1325

Vascular architecture and hypoxic profiles in human head and neck squamous cell carcinomas

K I E M Wijffels 1,2, J H A M Kaanders 1, P F J W Rijken 1, J Bussink 1, F J A van den Hoogen 2, H A M Marres 2, P C M de Wilde 3, J A Raleigh 4, A J van der Kogel 1
PMCID: PMC2363504  PMID: 10944611

Abstract

Tumour oxygenation and vasculature are determinants for radiation treatment outcome and prognosis in patients with squamous cell carcinomas of the head and neck. In this study we visualized and quantified these factors which may provide a predictive tool for new treatments. Twenty-one patients with stage III–IV squamous cell carcinomas of the head and neck were intravenously injected with pimonidazole, a bioreductive hypoxic marker. Tumour biopsies were taken 2 h later. Frozen tissue sections were stained for vessels and hypoxia by fluorescent immunohistochemistry. Twenty-two sections of biopsies of different head and neck sites were scanned and analysed with a computerized image analysis system. The hypoxic fractions varied from 0.02 to 0.29 and were independent from T- and N-classification, localization and differentiation grade. No significant correlation between hypoxic fraction and vascular density was observed. As a first attempt to categorize tumours based on their hypoxic profile, three different hypoxia patterns are described. The first category comprised tumours with large hypoxic, but viable, areas at distances even greater than 200 μm from the vessels. The second category showed a typical band-like distribution of hypoxia at an intermediate distance (50–200 μm) from the vessels with necrosis at greater distances. The third category demonstrated hypoxia already within 50 μm from the vessels, suggestive for acute hypoxia. This method of multiparameter analysis proved to be clinically feasible. The information on architectural patterns and the differences that exist between tumours can improve our understanding of the tumour micro-environment and may in the future be of assistance with the selection of (oxygenation modifying) treatment strategies. © 2000 Cancer Research Campaign

Keywords: pimonidazole, hypoxia, vascular density, head and neck carcinomas

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Selected References

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  1. Albo D., Granick M. S., Jhala N., Atkinson B., Solomon M. P. The relationship of angiogenesis to biological activity in human squamous cell carcinomas of the head and neck. Ann Plast Surg. 1994 Jun;32(6):588–594. doi: 10.1097/00000637-199406000-00005. [DOI] [PubMed] [Google Scholar]
  2. Azuma C., Raleigh J. A., Thrall D. E. Longevity of pimonidazole adducts in spontaneous canine tumors as an estimate of hypoxic cell lifetime. Radiat Res. 1997 Jul;148(1):35–42. [PubMed] [Google Scholar]
  3. Brizel D. M., Sibley G. S., Prosnitz L. R., Scher R. L., Dewhirst M. W. Tumor hypoxia adversely affects the prognosis of carcinoma of the head and neck. Int J Radiat Oncol Biol Phys. 1997 May 1;38(2):285–289. doi: 10.1016/s0360-3016(97)00101-6. [DOI] [PubMed] [Google Scholar]
  4. Burian M., Quint C., Neuchrist C. Angiogenic factors in laryngeal carcinomas: do they have prognostic relevance? Acta Otolaryngol. 1999 Mar;119(2):289–292. doi: 10.1080/00016489950181846. [DOI] [PubMed] [Google Scholar]
  5. Bussink J., Kaanders J. H., Rijken P. F., Martindale C. A., van der Kogel A. J. Multiparameter analysis of vasculature, perfusion and proliferation in human tumour xenografts. Br J Cancer. 1998;77(1):57–64. doi: 10.1038/bjc.1998.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bussink J., Kaanders J. H., Rijken P. F., Peters J. P., Hodgkiss R. J., Marres H. A., van der Kogel A. J. Vascular architecture and microenvironmental parameters in human squamous cell carcinoma xenografts: effects of carbogen and nicotinamide. Radiother Oncol. 1999 Feb;50(2):173–184. doi: 10.1016/s0167-8140(99)00010-9. [DOI] [PubMed] [Google Scholar]
  7. Dray T. G., Hardin N. J., Sofferman R. A. Angiogenesis as a prognostic marker in early head and neck cancer. Ann Otol Rhinol Laryngol. 1995 Sep;104(9 Pt 1):724–729. doi: 10.1177/000348949510400911. [DOI] [PubMed] [Google Scholar]
  8. Fenton B. M., Paoni S. F., Lee J., Koch C. J., Lord E. M. Quantification of tumour vasculature and hypoxia by immunohistochemical staining and HbO2 saturation measurements. Br J Cancer. 1999 Feb;79(3-4):464–471. doi: 10.1038/sj.bjc.6690072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. GRAY L. H., CONGER A. D., EBERT M., HORNSEY S., SCOTT O. C. The concentration of oxygen dissolved in tissues at the time of irradiation as a factor in radiotherapy. Br J Radiol. 1953 Dec;26(312):638–648. doi: 10.1259/0007-1285-26-312-638. [DOI] [PubMed] [Google Scholar]
  10. Gasparini G., Weidner N., Maluta S., Pozza F., Boracchi P., Mezzetti M., Testolin A., Bevilacqua P. Intratumoral microvessel density and p53 protein: correlation with metastasis in head-and-neck squamous-cell carcinoma. Int J Cancer. 1993 Nov 11;55(5):739–744. doi: 10.1002/ijc.2910550507. [DOI] [PubMed] [Google Scholar]
  11. Gatenby R. A., Kessler H. B., Rosenblum J. S., Coia L. R., Moldofsky P. J., Hartz W. H., Broder G. J. Oxygen distribution in squamous cell carcinoma metastases and its relationship to outcome of radiation therapy. Int J Radiat Oncol Biol Phys. 1988 May;14(5):831–838. doi: 10.1016/0360-3016(88)90002-8. [DOI] [PubMed] [Google Scholar]
  12. Janot F., Klijanienko J., Russo A., Mamet J. P., de Braud F., El-Naggar A. K., Pignon J. P., Luboinski B., Cvitkovic E. Prognostic value of clinicopathological parameters in head and neck squamous cell carcinoma: a prospective analysis. Br J Cancer. 1996 Feb;73(4):531–538. doi: 10.1038/bjc.1996.92. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jenssen N., Boysen M., Kjaerheim A., Bryne M. Low vascular density indicates poor response to radiotherapy in small glottic carcinomas. Pathol Res Pract. 1996 Nov;192(11):1090–1094. doi: 10.1016/S0344-0338(96)80024-0. [DOI] [PubMed] [Google Scholar]
  14. Kaanders J. H., Pop L. A., Marres H. A., van der Maazen R. W., van der Kogel A. J., van Daal W. A. Radiotherapy with carbogen breathing and nicotinamide in head and neck cancer: feasibility and toxicity. Radiother Oncol. 1995 Dec;37(3):190–198. doi: 10.1016/0167-8140(95)01660-0. [DOI] [PubMed] [Google Scholar]
  15. Kennedy A. S., Raleigh J. A., Perez G. M., Calkins D. P., Thrall D. E., Novotny D. B., Varia M. A. Proliferation and hypoxia in human squamous cell carcinoma of the cervix: first report of combined immunohistochemical assays. Int J Radiat Oncol Biol Phys. 1997 Mar 1;37(4):897–905. doi: 10.1016/s0360-3016(96)00539-1. [DOI] [PubMed] [Google Scholar]
  16. Leedy D. A., Trune D. R., Kronz J. D., Weidner N., Cohen J. I. Tumor angiogenesis, the p53 antigen, and cervical metastasis in squamous carcinoma of the tongue. Otolaryngol Head Neck Surg. 1994 Oct;111(4):417–422. doi: 10.1177/019459989411100405. [DOI] [PubMed] [Google Scholar]
  17. Lyng H., Sundfør K., Rofstad E. K. Oxygen tension in human tumours measured with polarographic needle electrodes and its relationship to vascular density, necrosis and hypoxia. Radiother Oncol. 1997 Aug;44(2):163–169. doi: 10.1016/s0167-8140(97)01920-8. [DOI] [PubMed] [Google Scholar]
  18. Moriyama M., Kumagai S., Kawashiri S., Kojima K., Kakihara K., Yamamoto E. Immunohistochemical study of tumour angiogenesis in oral squamous cell carcinoma. Oral Oncol. 1997 Sep;33(5):369–374. doi: 10.1016/s1368-8375(97)00025-0. [DOI] [PubMed] [Google Scholar]
  19. Murray J. D., Carlson G. W., McLaughlin K., Pennington M., Lynn M., DeRose P. B., Williams J. K., Cohen C. Tumor angiogenesis as a prognostic factor in laryngeal cancer. Am J Surg. 1997 Nov;174(5):523–526. doi: 10.1016/s0002-9610(97)00168-2. [DOI] [PubMed] [Google Scholar]
  20. Nordsmark M., Overgaard M., Overgaard J. Pretreatment oxygenation predicts radiation response in advanced squamous cell carcinoma of the head and neck. Radiother Oncol. 1996 Oct;41(1):31–39. doi: 10.1016/s0167-8140(96)91811-3. [DOI] [PubMed] [Google Scholar]
  21. Olive P. L., Johnston P. J., Banáth J. P., Durand R. E. The comet assay: a new method to examine heterogeneity associated with solid tumors. Nat Med. 1998 Jan;4(1):103–105. doi: 10.1038/nm0198-103. [DOI] [PubMed] [Google Scholar]
  22. Raleigh J. A., Calkins-Adams D. P., Rinker L. H., Ballenger C. A., Weissler M. C., Fowler W. C., Jr, Novotny D. B., Varia M. A. Hypoxia and vascular endothelial growth factor expression in human squamous cell carcinomas using pimonidazole as a hypoxia marker. Cancer Res. 1998 Sep 1;58(17):3765–3768. [PubMed] [Google Scholar]
  23. Raleigh J. A., Chou S. C., Arteel G. E., Horsman M. R. Comparisons among pimonidazole binding, oxygen electrode measurements, and radiation response in C3H mouse tumors. Radiat Res. 1999 May;151(5):580–589. [PubMed] [Google Scholar]
  24. Rijken P. F., Bernsen H. J., van der Kogel A. J. Application of an image analysis system to the quantitation of tumor perfusion and vascularity in human glioma xenografts. Microvasc Res. 1995 Sep;50(2):141–153. doi: 10.1006/mvre.1995.1048. [DOI] [PubMed] [Google Scholar]
  25. Salven P., Heikkilä P., Anttonen A., Kajanti M., Joensuu H. Vascular endothelial growth factor in squamous cell head and neck carcinoma: expression and prognostic significance. Mod Pathol. 1997 Nov;10(11):1128–1133. [PubMed] [Google Scholar]
  26. Schlingemann R. O., Dingjan G. M., Emeis J. J., Blok J., Warnaar S. O., Ruiter D. J. Monoclonal antibody PAL-E specific for endothelium. Lab Invest. 1985 Jan;52(1):71–76. [PubMed] [Google Scholar]
  27. Shpitzer T., Chaimoff M., Gal R., Stern Y., Feinmesser R., Segal K. Tumor angiogenesis as a prognostic factor in early oral tongue cancer. Arch Otolaryngol Head Neck Surg. 1996 Aug;122(8):865–868. doi: 10.1001/archotol.1996.01890200055012. [DOI] [PubMed] [Google Scholar]
  28. THOMLINSON R. H., GRAY L. H. The histological structure of some human lung cancers and the possible implications for radiotherapy. Br J Cancer. 1955 Dec;9(4):539–549. doi: 10.1038/bjc.1955.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Varia M. A., Calkins-Adams D. P., Rinker L. H., Kennedy A. S., Novotny D. B., Fowler W. C., Jr, Raleigh J. A. Pimonidazole: a novel hypoxia marker for complementary study of tumor hypoxia and cell proliferation in cervical carcinoma. Gynecol Oncol. 1998 Nov;71(2):270–277. doi: 10.1006/gyno.1998.5163. [DOI] [PubMed] [Google Scholar]
  30. Zätterström U. K., Brun E., Willén R., Kjellén E., Wennerberg J. Tumor angiogenesis and prognosis in squamous cell carcinoma of the head and neck. Head Neck. 1995 Jul-Aug;17(4):312–318. doi: 10.1002/hed.2880170407. [DOI] [PubMed] [Google Scholar]

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