Comparison of immunostainings for proliferating cell nuclear antigen and Ki-67 in human extraocular lesions

  • Tatsuo Kodama
  • Katsue Kawamoto
  • Tatsuro Kono
  • Yuzo Shibuya
  • Tomoichi Setogawa
Clinical Investigation


• Background: Cell kinetic information is helpful to understand disease progression, treatment response, and prognosis of the neoplasms. To compare the usefulness and limitations of antibodies that recognize the cell cycle-associated molecules, proliferating cell nuclear antigen (PCNA) and Ki-67, we performed immunostaining in formalin-fixed, paraffin-embedded tissue sections of extraocular lesions. • Methods: Specimens were obtained from patients undergoing routine surgical procedures. Formalin-fixed, paraffin-embedded sections were stained for PCNA and Ki-67 using the monoclonal antibodies PC10 and MIB-1, respectively. Microwave oven heating for antigen retrieval was performed before immunostaining. • Results: In squamous cell carcinomas and basal cell carcinomas, PCNA immunostaining varied greatly. Basal cells of benign epithelial lesions showed moderate to weak PCNA staining. Strong PCNA immunoreactivity was demonstrated in foci of inflammation and germinal centers. Microwave processing enhanced the intensity of those PCNA immunostainings. As the PCNA immunostaining intensity had a great variability, the absolute numbers of PCNA-positive cells were hard to count in some tissues. High Ki-67 counts were observed in squamous cell carcinomas, foci of inflammatory cells, and germinal center cells. Basal cell carcinomas and benign lesions showed low Ki-67 counts. Every section showed clear nuclear staining in Ki-67 immunostaining. • Conclusion: Careful consideration is required in the assessment of cell proliferation using PCNA. The immunostaining of Ki-67 may be more accurate than that of PCNA for evaluating cell proliferation in formalin-fixed, paraffin-embedded tissues.


Squamous Cell Carcinoma Proliferate Cell Nuclear Antigen Basal Cell Carcinoma Germinal Center Microwave Processing 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Bravo R, Macdonald-Bravo H (1987) Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites. J Cell Biol 105: 1549–1554CrossRefPubMedGoogle Scholar
  2. 2.
    Bravo R, Frank R, Blundell PA, MacDonald-Bravo H (1987) Cyclin-PCNA is thé auxiliary protein of DNA polymerase-δ. Nature 326: 515–517CrossRefPubMedGoogle Scholar
  3. 3.
    Cattoretti G, Becker MHG, Key G, Duchrow M, Schlüter C, Galle J, Gerdes J (1992) Monoclonal antibodies against recombinant parts of the Ki-67 antigen (MIB 1 and MIB 3) detect proliferating cells in microwave-processed formalinfixed paraffin sections. J Pathol 168: 357–363CrossRefPubMedGoogle Scholar
  4. 4.
    Dawson AE, Norton JA, Weinberg DS (1990) Comparative assessment of Proliferation and DNA content in breast carcinoma by image analysis and flow cytometry. Am J Pathol 136: 1115–1124PubMedGoogle Scholar
  5. 5.
    Gan L, van Setten G, Fagerholm P (1995) Proliferating cell nuclear antigen: contradictory results regarding its presence in the lens. Graefe's Arch Clin Exp Ophthalmol 233: 792–794CrossRefGoogle Scholar
  6. 6.
    Gerdes J, Li L, Schlueter C, Duchrow M, Wohlenberg C, Gerlach C, Stahmer I, Kloth S, Brandt E, Flad H-D (1991) Immunobiochemical and molecular biologie characterization of the cell proliferation-associated nuclear antigen that is defined by monoclonal antibody Ki-67. Am J Pathol 138: 867–873PubMedGoogle Scholar
  7. 7.
    Ghazvini S, Kroll S, Char DH, Frigillana H (1995) Comparative analysis of proliferating cell nuclear antigen, bromodeoxyuridine, and mitotic index in uveal melanoma. Invest Ophthalmol Vis Sci 36: 2762–2767PubMedGoogle Scholar
  8. 8.
    Hall PA, Levison DA (1990) Assessment of cell proliferation in histological material. J Clin Pathol 43: 184–192PubMedGoogle Scholar
  9. 9.
    Hall PA, Levison DA, Woods AL, Yu CC-W, Kellock DB, Watkins JA, Barnes DM, Gillett CE, Camplejohn R, Dover R, Waseem NH, Lane DP (1990) Proliferating cell nuclear antigen (PCNA) immunolocalization in paraffin sections: an index of cell proliferation with evidence of deregulated expression in some neoplasms. J Pathol 162: 285–294CrossRefPubMedGoogle Scholar
  10. 10.
    Hergott GJ, Kalnins VI (1991) Expression of proliferating cell nuclear antigen in migrating retinal pigment epithelial cells during wound healing in organ culture. Exp Cell Res 195: 307–314CrossRefPubMedGoogle Scholar
  11. 11.
    Hodge WG, Duclos AJ, Rocha G, Antecka E, Baines MG, Corriveau C, Brownstein S, Deschenes J (1995) DNA index and S phase fraction in uveal malignant melanomas. Br J Ophthalmol 79:521–526PubMedGoogle Scholar
  12. 12.
    Kamel OW, Franklin WA, Ringus JC, Neyer JS (1989) Thymidine labeling index and Ki-67 growth fraction in lesions of the breast. Am J Pathol 134: 107–113PubMedGoogle Scholar
  13. 13.
    Kindy-Degnan N, Char DH, Swift P, Kaleta S, Ljung BM (1989) Bromodeoxyuridine uptake in the assessment of hyperthermie therapy for intraocular tumon. Arch Ophthalmol 107: 746–750PubMedGoogle Scholar
  14. 14.
    Mathews MB, Bernstein RM, Franza BR, Garrels Jl (1984) Identity of the proliferating cell nuclear antigen and cyclin. Nature 309: 374–376CrossRefPubMedGoogle Scholar
  15. 15.
    McCormick D, Hall PA (1992) The complexities of proliferating cell nuclear antigen. Histopathology 21: 591–594PubMedGoogle Scholar
  16. 16.
    Mooy CM, de Jong PTVM, Van der Kwast TH, Hulder PGH, Jager MI, Ruiter DJ (1990) Ki-67 immunostaining in uveal melanoma. The effect of preenucleation radiotherapy. Ophthalmology 97: 1275–1280PubMedGoogle Scholar
  17. 17.
    Murata T, Ishibashi T, Inomata H (1993) Localizations of epidermal growth factor receptor and proliferating cell nuclear antigen during corneal wound healing. Graefe's Arch Clin Exp Ophthalmol 231: 104–108Google Scholar
  18. 18.
    Pe'er J, Gnessin H, Shargal Y, Livini N (1994) PC-10 immunostaining of proliferating cell nuclear antigen in posterior uveal melanoma: enucleation versus enucleation postirradiation groups. Ophthalmology 101: 56–62PubMedGoogle Scholar
  19. 19.
    Schrape S, Jones DB, Wright DH (1987) A comparison of three methods for the determination of the growth fraction in non-Hodgkin's lymphoma. Br J Cancer 55: 283–286PubMedGoogle Scholar
  20. 20.
    Shi S-R, Key ME, Kalra KL (1991) Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections. J Histochem Cytochem 39: 741–748PubMedGoogle Scholar
  21. 21.
    Start RD, Cross SS, Clelland C, Silcocks PB, Rogers K, Smith JHF (1992) Delay in fixation does not affect the immunoreactivity of proliferating cell nuclear antigen (PCNA). J Pathol 168: 197–199CrossRefPubMedGoogle Scholar
  22. 22.
    Suzuki K, Katoh R, Kawaoi A (1992) Immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA) in formalin-fixed, paraffin-embedded sections from rat and human tissues. Acta Histochem Cytochem 25: 13–21Google Scholar
  23. 23.
    Walker RA, Camplejohn RS (1988) Comparison of monoclonal antibody Ki-67 reactivity with grade and DNA flow cytometry of breast carcinomas. Br J Cancer 57: 281–283PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1997

Authors and Affiliations

  • Tatsuo Kodama
    • 1
  • Katsue Kawamoto
    • 1
  • Tatsuro Kono
    • 1
  • Yuzo Shibuya
    • 1
  • Tomoichi Setogawa
    • 1
  1. 1.Department of OphthalmologyShimane Medical UniversityIzumo, ShimaneJapan

Personalised recommendations