Skip to main content
NCBI home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1990 Jun;61(6):821–827. doi: 10.1038/bjc.1990.184

An immunohistochemical assessment of cellular proliferation markers in head and neck squamous cell cancers.

J H Kearsley 1, K L Furlong 1, R A Cooke 1, M J Waters 1
PMCID: PMC1971683  PMID: 2372483

Abstract

Prognostic information is essential for the evaluation, judgement and optimal treatment of patients with squamous cell cancers (SCCs) of the upper aerodigestive tract. Using immunohistochemical and flow cytometric techniques, we have studied the significance of cellular expression of the Ki-67 antigen, epidermal growth factor receptor (EGFR), the transferrin receptor (TFR) and DNA ploidy status in a prospective analysis of patients with SCCs of the head and neck region. All 42 fresh tumour samples (five well differentiated; 28 moderately differentiated; nine poorly differentiated) expressed both EGFR and TFR to varying degrees. Receptor expression was most marked on the peripheral invading margin of cancer cell islands although staining was also demonstrated in a random fashion within cellular islands and consistently along the basal cell layer of overlying stratified squamous epithelium. The percentage of cancer cells that reacted with the Ki-67 monoclonal antibody was assessed as low (less than 10%) in 15 samples (35.8%), intermediate (10-30%) in 19 samples (45.2%) and high (greater than 30%) in eight samples (19.0%). Eleven of 15 samples (73%) with a low percentage reactivity were DNA diploid, whereas seven of eight samples (87.5%) with a high percentage reactivity were DNA aneuploid. Poorly differentiated SCCs were significantly more often aneuploid than were either moderately or well differentiated tumours. Our results suggest that EGFR and TFR are widely distributed on SCCs, especially on proliferating cells at the invading tumour margin. In addition, there is a close spatial correlation between cells expressing EGFR, TFR and those expressing the Ki-67 antigen. Tumours in which the staining intensity for both EGFR and TFR was intense invariably expressed the Ki-67 antigen in a high proportion of cells. Further patient follow-up will be important in determining whether intense EGFR and TFR staining, combined with a high percentage reactivity with Ki-67 antibody and DNA aneuploidy, will ultimately define a subset of head and neck cancer patients with a poor clinical outcome.

Full text

PDF
821

Images in this article

824Figure 1
on p.824
825Figure 2
on p.825
825Figure 4
on p.825

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bouzubar N., Walker K. J., Griffiths K., Ellis I. O., Elston C. W., Robertson J. F., Blamey R. W., Nicholson R. I. Ki67 immunostaining in primary breast cancer: pathological and clinical associations. Br J Cancer. 1989 Jun;59(6):943–947. doi: 10.1038/bjc.1989.200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brown D. C., Cole D., Gatter K. C., Mason D. Y. Carcinoma of the cervix uteri: an assessment of tumour proliferation using the monoclonal antibody Ki67. Br J Cancer. 1988 Feb;57(2):178–181. doi: 10.1038/bjc.1988.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Castagnola J., MacLeod C., Sunada H., Mendelsohn J., Taetle R. Effects of epidermal growth factor on transferrin receptor phosphorylation and surface expression in malignant epithelial cells. J Cell Physiol. 1987 Sep;132(3):492–500. doi: 10.1002/jcp.1041320311. [DOI] [PubMed] [Google Scholar]
  4. Chauvel P., Courdi A., Gioanni J., Vallicioni J., Santini J., Demard F. The labelling index: a prognostic factor in head and neck carcinoma. Radiother Oncol. 1989 Mar;14(3):231–237. doi: 10.1016/0167-8140(89)90171-0. [DOI] [PubMed] [Google Scholar]
  5. Derynck R., Goeddel D. V., Ullrich A., Gutterman J. U., Williams R. D., Bringman T. S., Berger W. H. Synthesis of messenger RNAs for transforming growth factors alpha and beta and the epidermal growth factor receptor by human tumors. Cancer Res. 1987 Feb 1;47(3):707–712. [PubMed] [Google Scholar]
  6. Earp H. S., Austin K. S., Blaisdell J., Rubin R. A., Nelson K. G., Lee L. W., Grisham J. W. Epidermal growth factor (EGF) stimulates EGF receptor synthesis. J Biol Chem. 1986 Apr 15;261(11):4777–4780. [PubMed] [Google Scholar]
  7. Fitzpatrick S. L., Brightwell J., Wittliff J. L., Barrows G. H., Schultz G. S. Epidermal growth factor binding by breast tumor biopsies and relationship to estrogen receptor and progestin receptor levels. Cancer Res. 1984 Aug;44(8):3448–3453. [PubMed] [Google Scholar]
  8. Friedlander M. L., Hedley D. W., Taylor I. W., Russell P., Coates A. S., Tattersall M. H. Influence of cellular DNA content on survival in advanced ovarian cancer. Cancer Res. 1984 Jan;44(1):397–400. [PubMed] [Google Scholar]
  9. Gatter K. C., Brown G., Trowbridge I. S., Woolston R. E., Mason D. Y. Transferrin receptors in human tissues: their distribution and possible clinical relevance. J Clin Pathol. 1983 May;36(5):539–545. doi: 10.1136/jcp.36.5.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gerdes J., Dallenbach F., Lennert K., Lemke H., Stein H. Growth fractions in malignant non-Hodgkin's lymphomas (NHL) as determined in situ with the monoclonal antibody Ki-67. Hematol Oncol. 1984 Oct-Dec;2(4):365–371. doi: 10.1002/hon.2900020406. [DOI] [PubMed] [Google Scholar]
  11. Gerdes J., Lelle R. J., Pickartz H., Heidenreich W., Schwarting R., Kurtsiefer L., Stauch G., Stein H. Growth fractions in breast cancers determined in situ with monoclonal antibody Ki-67. J Clin Pathol. 1986 Sep;39(9):977–980. doi: 10.1136/jcp.39.9.977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gerdes J., Schwab U., Lemke H., Stein H. Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. Int J Cancer. 1983 Jan 15;31(1):13–20. doi: 10.1002/ijc.2910310104. [DOI] [PubMed] [Google Scholar]
  13. Goldsmith M. M., Cresson D. H., Arnold L. A., Postma D. S., Askin F. B., Pillsbury H. C. DNA flow cytometry as a prognostic indicator in head and neck cancer. Otolaryngol Head Neck Surg. 1987 Apr;96(4):307–318. doi: 10.1177/019459988709600402. [DOI] [PubMed] [Google Scholar]
  14. Hedley D. W., Friedlander M. L., Taylor I. W., Rugg C. A., Musgrove E. A. Method for analysis of cellular DNA content of paraffin-embedded pathological material using flow cytometry. J Histochem Cytochem. 1983 Nov;31(11):1333–1335. doi: 10.1177/31.11.6619538. [DOI] [PubMed] [Google Scholar]
  15. Hedley D. W., Rugg C. A., Gelber R. D. Association of DNA index and S-phase fraction with prognosis of nodes positive early breast cancer. Cancer Res. 1987 Sep 1;47(17):4729–4735. [PubMed] [Google Scholar]
  16. Hendler F. J., Ozanne B. W. Human squamous cell lung cancers express increased epidermal growth factor receptors. J Clin Invest. 1984 Aug;74(2):647–651. doi: 10.1172/JCI111463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hsu S. M., Raine L., Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  18. Johnson T. S., Williamson K. D., Cramer M. M., Peters L. J. Flow cytometric analysis of head and neck carcinoma DNA index and S-fraction from paraffin-embedded sections: comparison with malignancy grading. Cytometry. 1985 Sep;6(5):461–470. doi: 10.1002/cyto.990060511. [DOI] [PubMed] [Google Scholar]
  19. King L. E., Jr What does epidermal growth factor do and how does it do it? J Invest Dermatol. 1985 Mar;84(3):165–167. doi: 10.1111/1523-1747.ep12264680. [DOI] [PubMed] [Google Scholar]
  20. Marks F. What's new in oncogenes and growth factors? Pathol Res Pract. 1987 Dec;182(6):831–848. doi: 10.1016/S0344-0338(87)80050-X. [DOI] [PubMed] [Google Scholar]
  21. McGurrin J. F., Doria M. I., Jr, Dawson P. J., Karrison T., Stein H. O., Franklin W. A. Assessment of tumor cell kinetics by immunohistochemistry in carcinoma of breast. Cancer. 1987 May 15;59(10):1744–1750. doi: 10.1002/1097-0142(19870515)59:10<1744::aid-cncr2820591012>3.0.co;2-d. [DOI] [PubMed] [Google Scholar]
  22. Merkel D. E., Dressler L. G., McGuire W. L. Flow cytometry, cellular DNA content, and prognosis in human malignancy. J Clin Oncol. 1987 Oct;5(10):1690–1703. doi: 10.1200/JCO.1987.5.10.1690. [DOI] [PubMed] [Google Scholar]
  23. Merlino G. T., Xu Y. H., Richert N., Clark A. J., Ishii S., Banks-Schlegel S., Pastan I. Elevated epidermal growth factor receptor gene copy number and expression in a squamous carcinoma cell line. J Clin Invest. 1985 Mar;75(3):1077–1079. doi: 10.1172/JCI111770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Neal D. E., Marsh C., Bennett M. K., Abel P. D., Hall R. R., Sainsbury J. R., Harris A. L. Epidermal-growth-factor receptors in human bladder cancer: comparison of invasive and superficial tumours. Lancet. 1985 Feb 16;1(8425):366–368. doi: 10.1016/s0140-6736(85)91386-8. [DOI] [PubMed] [Google Scholar]
  25. Osborene C. K., Hamilton B., Nover M. Receptor binding and processing of epidermal growth factor by human breast cancer cells. J Clin Endocrinol Metab. 1982 Jul;55(1):86–93. doi: 10.1210/jcem-55-1-86. [DOI] [PubMed] [Google Scholar]
  26. Sainsbury J. R., Farndon J. R., Sherbet G. V., Harris A. L. Epidermal-growth-factor receptors and oestrogen receptors in human breast cancer. Lancet. 1985 Feb 16;1(8425):364–366. doi: 10.1016/s0140-6736(85)91385-6. [DOI] [PubMed] [Google Scholar]
  27. Schneiderman M. A. Epidemiology, carcinogenesis and virology: time trends: United States 1953-73. Laryngoscope. 1978 Jan;88(1 Pt 2 Suppl 8):44–49. [PubMed] [Google Scholar]
  28. Shepherd N. A., Richman P. I., England J. Ki-67 derived proliferative activity in colorectal adenocarcinoma with prognostic correlations. J Pathol. 1988 Jul;155(3):213–219. doi: 10.1002/path.1711550306. [DOI] [PubMed] [Google Scholar]
  29. Shindelman J. E., Ortmeyer A. E., Sussman H. H. Demonstration of the transferrin receptor in human breast cancer tissue. Potential marker for identifying dividing cells. Int J Cancer. 1981 Mar 15;27(3):329–334. doi: 10.1002/ijc.2910270311. [DOI] [PubMed] [Google Scholar]
  30. Sledge G. W., Jr, Eble J. N., Roth B. J., Wuhrman B. P., Fineberg N., Einhorn L. H. Relation of proliferative activity to survival in patients with advanced germ cell cancer. Cancer Res. 1988 Jul 1;48(13):3864–3868. [PubMed] [Google Scholar]
  31. Stoscheck C. M., King L. E., Jr Role of epidermal growth factor in carcinogenesis. Cancer Res. 1986 Mar;46(3):1030–1037. [PubMed] [Google Scholar]
  32. Sutherland R., Delia D., Schneider C., Newman R., Kemshead J., Greaves M. Ubiquitous cell-surface glycoprotein on tumor cells is proliferation-associated receptor for transferrin. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4515–4519. doi: 10.1073/pnas.78.7.4515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Trowbridge I. S., Omary M. B. Human cell surface glycoprotein related to cell proliferation is the receptor for transferrin. Proc Natl Acad Sci U S A. 1981 May;78(5):3039–3043. doi: 10.1073/pnas.78.5.3039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tubiana M., Courdi A. Cell proliferation kinetics in human solid tumors: relation to probability of metastatic dissemination and long-term survival. Radiother Oncol. 1989 May;15(1):1–18. doi: 10.1016/0167-8140(89)90113-8. [DOI] [PubMed] [Google Scholar]
  35. Tubiana M., Pejovic M. H., Koscielny S., Chavaudra N., Malaise E. Growth rate, kinetics of tumor cell proliferation and long-term outcome in human breast cancer. Int J Cancer. 1989 Jul 15;44(1):17–22. doi: 10.1002/ijc.2910440104. [DOI] [PubMed] [Google Scholar]
  36. Veale D., Ashcroft T., Marsh C., Gibson G. J., Harris A. L. Epidermal growth factor receptors in non-small cell lung cancer. Br J Cancer. 1987 May;55(5):513–516. doi: 10.1038/bjc.1987.104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Viac J., Chardonnet Y., Bouvard V., Leval J., Morgon A., Thivolet J. Virus expression. EGF and transferrin receptors in human papillomas. Virchows Arch A Pathol Anat Histopathol. 1987;411(1):73–77. doi: 10.1007/BF00734517. [DOI] [PubMed] [Google Scholar]
  38. Wada H. G., Hass P. E., Sussman H. H. Transferrin receptor in human placental brush border membranes. Studies on the binding of transferrin to placental membrane vesicles and the identification of a placental brush border glycoprotein with high affinity for transferrin. J Biol Chem. 1979 Dec 25;254(24):12629–12635. [PubMed] [Google Scholar]
  39. Walker R. A., Camplejohn R. S. DNA flow cytometry of human breast carcinomas and its relationship to transferrin and epidermal growth factor receptors. J Pathol. 1986 Sep;150(1):37–42. doi: 10.1002/path.1711500107. [DOI] [PubMed] [Google Scholar]
  40. Waterfield M. D., Mayes E. L., Stroobant P., Bennet P. L., Young S., Goodfellow P. N., Banting G. S., Ozanne B. A monoclonal antibody to the human epidermal growth factor receptor. J Cell Biochem. 1982;20(2):149–161. doi: 10.1002/jcb.240200207. [DOI] [PubMed] [Google Scholar]
  41. Wiley H. S., Kaplan J. Epidermal growth factor rapidly induces a redistribution of transferrin receptor pools in human fibroblasts. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7456–7460. doi: 10.1073/pnas.81.23.7456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Williams M. L., Nettesheim P. Lung colony assay with a squamous cell carcinoma derived from the respiratory tract of mice. J Natl Cancer Inst. 1973 Nov;51(5):1513–1520. doi: 10.1093/jnci/51.5.1513. [DOI] [PubMed] [Google Scholar]
  43. Wrba F., Chott A., Reiner A., Reiner G., Markis-Ritzinger E., Holzner J. H. Ki-67 immunoreactivity in breast carcinomas in relation to transferrin receptor expression, estrogen receptor status and morphological criteria. An immunohistochemical study. Oncology. 1989;46(4):255–259. doi: 10.1159/000226727. [DOI] [PubMed] [Google Scholar]
  44. Yamamoto T., Kamata N., Kawano H., Shimizu S., Kuroki T., Toyoshima K., Rikimaru K., Nomura N., Ishizaki R., Pastan I. High incidence of amplification of the epidermal growth factor receptor gene in human squamous carcinoma cell lines. Cancer Res. 1986 Jan;46(1):414–416. [PubMed] [Google Scholar]
  45. Young G. A., Hedley D. W., Rugg C. A., Iland H. J. The prognostic significance of proliferative activity in poor histology non-Hodgkin's lymphoma: a flow cytometry study using archival material. Eur J Cancer Clin Oncol. 1987 Oct;23(10):1497–1504. doi: 10.1016/0277-5379(87)90092-7. [DOI] [PubMed] [Google Scholar]
  46. Zarbo R. J., Crissman J. D. The surgical pathology of head and neck cancer. Semin Oncol. 1988 Feb;15(1):10–19. [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES