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. 1994 Jan;47(1):18–22. doi: 10.1136/jcp.47.1.18

MIB-1, Ki67, and PCNA scores and DNA flow cytometry in intermediate grade malignant lymphomas.

A Pich 1, R Ponti 1, G Valente 1, L Chiusa 1, M Geuna 1, D Novero 1, G Palestro 1
PMCID: PMC501749  PMID: 7907607

Abstract

AIMS--To verify the correlation between MIB-1, Ki67, and proliferating cell nuclear antigen (PCNA-PC10) scores and S-phase fraction in intermediate grade non-Hodgkin's lymphomas (Working Formulation F); and their reliability in differently processed tissues. METHODS--Forty one non-Hodgkin's lymphomas were classified as (F) intermediate grade malignant lymphomas according to the Working Formulation; mitotic counts and percentage of large cells were assessed for each case. Sections from formalin fixed, paraffin wax embedded tissues were stained with anti MIB-1 monoclonal antibody, after microwave oven processing, and anti-PCNA (PC10) monoclonal antibody using an avidin-biotin immunoperoxidase (ABC) method. One thousand cells from 10 representative fields were scored. Frozen sections from surgical specimens were stained with Ki67 monoclonal antibody using the ABC method; the fraction of Ki67 positive cells was calculated scoring 1000 cells. Flow cytometry analysis (FCM) was performed on cell suspensions from fresh tissues. Correlations between data were estimated using linear regression. RESULTS--A linear correlation was found between MIB-1 and Ki67 scores (r = 0.92; p < 0.00001); between MIB-1 and PCNA scores (r = 0.79; p < 0.00001); and between MIB-1 score and S-phase fraction (r = 0.51; p = 0.0006). A linear correlation was also found between Ki67 and PCNA scores (r = 0.85; p < 0.00001); between Ki67 score and S-phase fraction (r = 0.6; p = 0.0002); and between PCNA score and S-phase fraction (r = 0.74; p < 0.00001). A correlation was found between mitotic counts and MIB-1 (r = 0.56; p = 0.0001), PCNA (r = 0.51; p = 0.0007), or Ki67 scores (r = 0.47; p = 0.002); between the percentage of large cells and MIB-1 (r = 0.49; p = 0.0009), PCNA (r = 0.6; p = 0.00003), and Ki67 scores (r = 0.53; p = 0.0003) and S-phase fraction (r = 0.55; p = 0.0002). CONCLUSION--MIB-1, Ki67, and PCNA (PC10) scores and S-phase fraction are highly correlated and equally well represent the proliferative activity of intermediate grade non-Hodgkin's lymphomas in differently processed material. MIB-1 and PCNA stains can be applied even on small biopsy specimens. MIB-1 produces homogenous staining without background; it also strongly stains mitotic figures. It can be performed on routinely processed tissues, permitting the simultaneous evaluation of the morphology and tumour cell kinetics. The wide standard deviations of the proliferative indices found for intermediate grade NHL suggest that this category probably includes various degrees of malignancy.

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Selected References

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  1. Aaltomaa S., Lipponen P., Syrjänen K. Prognostic value of cell proliferation in breast cancer as determined by proliferating cell nuclear antigen (PCNA) immunostaining. Anticancer Res. 1992 Jul-Aug;12(4):1281–1286. [PubMed] [Google Scholar]
  2. Allegranza A., Girlando S., Arrigoni G. L., Veronese S., Mauri F. A., Gambacorta M., Pollo B., Dalla Palma P., Barbareschi M. Proliferating cell nuclear antigen expression in central nervous system neoplasms. Virchows Arch A Pathol Anat Histopathol. 1991;419(5):417–423. doi: 10.1007/BF01605076. [DOI] [PubMed] [Google Scholar]
  3. Braylan R. C., Diamond L. W., Powell M. L., Harty-Golder B. Percentage of cells in the S phase of the cell cycle in human lymphoma determined by flow cytometry. Cytometry. 1980 Nov;1(3):171–174. doi: 10.1002/cyto.990010302. [DOI] [PubMed] [Google Scholar]
  4. Burger P. C., Shibata T., Kleihues P. The use of the monoclonal antibody Ki-67 in the identification of proliferating cells: application to surgical neuropathology. Am J Surg Pathol. 1986 Sep;10(9):611–617. doi: 10.1097/00000478-198609000-00003. [DOI] [PubMed] [Google Scholar]
  5. Celis J. E., Bravo R., Larsen P. M., Fey S. J. Cyclin: a nuclear protein whose level correlates directly with the proliferative state of normal as well as transformed cells. Leuk Res. 1984;8(2):143–157. doi: 10.1016/0145-2126(84)90135-8. [DOI] [PubMed] [Google Scholar]
  6. Celis J. E., Celis A. Cell cycle-dependent variations in the distribution of the nuclear protein cyclin proliferating cell nuclear antigen in cultured cells: subdivision of S phase. Proc Natl Acad Sci U S A. 1985 May;82(10):3262–3266. doi: 10.1073/pnas.82.10.3262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Costa A., Mazzini G., Del Bino G., Silvestrini R. DNA content and kinetic characteristics of non-Hodgkin's lymphoma: determined by flow cytometry and autoradiography. Cytometry. 1981 Nov;2(3):185–188. doi: 10.1002/cyto.990020310. [DOI] [PubMed] [Google Scholar]
  8. Crocker J., Macartney J. C., Smith P. J. Correlation between DNA flow cytometric and nucleolar organizer region data in non-Hodgkin's lymphomas. J Pathol. 1988 Feb;154(2):151–156. doi: 10.1002/path.1711540207. [DOI] [PubMed] [Google Scholar]
  9. Crocker J. Proliferation indices in malignant lymphomas. Clin Exp Immunol. 1989 Sep;77(3):299–308. [PMC free article] [PubMed] [Google Scholar]
  10. Dawson A. E., Norton J. A., Weinberg D. S. Comparative assessment of proliferation and DNA content in breast carcinoma by image analysis and flow cytometry. Am J Pathol. 1990 May;136(5):1115–1124. [PMC free article] [PubMed] [Google Scholar]
  11. Diamond L. W., Braylan R. C. Flow analysis of DNA content and cell size in non-Hodgkin's lymphoma. Cancer Res. 1980 Mar;40(3):703–712. [PubMed] [Google Scholar]
  12. Diamond L. W., Nathwani B. N., Rappaport H. Flow cytometry in the diagnosis and classification of malignant lymphoma and leukemia. Cancer. 1982 Sep 15;50(6):1122–1135. doi: 10.1002/1097-0142(19820915)50:6<1122::aid-cncr2820500616>3.0.co;2-q. [DOI] [PubMed] [Google Scholar]
  13. Düe W., Dieckmann K. P., Loy V. Immunohistological determination of proliferative activity in seminomas. J Clin Pathol. 1988 Mar;41(3):304–307. doi: 10.1136/jcp.41.3.304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Garcia R. L., Coltrera M. D., Gown A. M. Analysis of proliferative grade using anti-PCNA/cyclin monoclonal antibodies in fixed, embedded tissues. Comparison with flow cytometric analysis. Am J Pathol. 1989 Apr;134(4):733–739. [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]
  18. Hall P. A., Levison D. A., Woods A. L., Yu C. C., Kellock D. B., Watkins J. A., Barnes D. M., Gillett C. E., Camplejohn R., Dover R. Proliferating cell nuclear antigen (PCNA) immunolocalization in paraffin sections: an index of cell proliferation with evidence of deregulated expression in some neoplasms. J Pathol. 1990 Dec;162(4):285–294. doi: 10.1002/path.1711620403. [DOI] [PubMed] [Google Scholar]
  19. Hall P. A., Richards M. A., Gregory W. M., d'Ardenne A. J., Lister T. A., Stansfeld A. G. The prognostic value of Ki67 immunostaining in non-Hodgkin's lymphoma. J Pathol. 1988 Mar;154(3):223–235. doi: 10.1002/path.1711540305. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Isola J., Kallioniemi O. P., Korte J. M., Wahlström T., Aine R., Helle M., Helin H. Steroid receptors and Ki-67 reactivity in ovarian cancer and in normal ovary: correlation with DNA flow cytometry, biochemical receptor assay, and patient survival. J Pathol. 1990 Dec;162(4):295–301. doi: 10.1002/path.1711620404. [DOI] [PubMed] [Google Scholar]
  22. Joensuu H., Klemi P. J. Comparison of nuclear DNA content in primary and metastatic differentiated thyroid carcinoma. Am J Clin Pathol. 1988 Jan;89(1):35–40. doi: 10.1093/ajcp/89.1.35. [DOI] [PubMed] [Google Scholar]
  23. Landberg G., Roos G. Expression of proliferating cell nuclear antigen (PCNA) and Ki-67 antigen in human malignant hematopoietic cells. Acta Oncol. 1991;30(8):917–921. doi: 10.3109/02841869109088244. [DOI] [PubMed] [Google Scholar]
  24. Lardelli P., Bookman M. A., Sundeen J., Longo D. L., Jaffe E. S. Lymphocytic lymphoma of intermediate differentiation. Morphologic and immunophenotypic spectrum and clinical correlations. Am J Surg Pathol. 1990 Aug;14(8):752–763. doi: 10.1097/00000478-199008000-00007. [DOI] [PubMed] [Google Scholar]
  25. Lipponen P. K., Eskelinen M. J. Cell proliferation of transitional cell bladder tumours determined by PCNA/cyclin immunostaining and its prognostic value. Br J Cancer. 1992 Jul;66(1):171–176. doi: 10.1038/bjc.1992.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mellon K., Neal D. E., Robinson M. C., Marsh C., Wright C. Cell cycling in bladder carcinoma determined by monoclonal antibody Ki67. Br J Urol. 1990 Sep;66(3):281–285. doi: 10.1111/j.1464-410x.1990.tb14927.x. [DOI] [PubMed] [Google Scholar]
  27. Merkel D. E., McGuire W. L. Ploidy, proliferative activity and prognosis. DNA flow cytometry of solid tumors. Cancer. 1990 Mar 1;65(5):1194–1205. doi: 10.1002/1097-0142(19900301)65:5<1194::aid-cncr2820650528>3.0.co;2-m. [DOI] [PubMed] [Google Scholar]
  28. Pich A., Chiusa L., Pisani P., Krengli M., Pia F., Navone R. Argyrophilic nucleolar organizer region counts and proliferating cell nuclear antigen scores are two reliable indicators of survival in pharyngeal carcinoma. J Cancer Res Clin Oncol. 1992;119(2):106–110. doi: 10.1007/BF01209665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pich A., Valente G., Azzoni L., Stramignoni A., Margaria E., Tasso M. Argyrophilic nucleolar organizer region counts and Ki-67 scores in human renal cell carcinoma. Pathol Res Pract. 1991 May;187(4):482–486. doi: 10.1016/s0344-0338(11)80011-7. [DOI] [PubMed] [Google Scholar]
  30. Raymond W. A., Leong A. S., Bolt J. W., Milios J., Jose J. S. Growth fractions in human prostatic carcinoma determined by Ki-67 immunostaining. J Pathol. 1988 Oct;156(2):161–167. doi: 10.1002/path.1711560211. [DOI] [PubMed] [Google Scholar]
  31. Schrape S., Jones D. B., Wright D. H. A comparison of three methods for the determination of the growth fraction in non-Hodgkin's lymphoma. Br J Cancer. 1987 Mar;55(3):283–286. doi: 10.1038/bjc.1987.54. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schultz H. B., Ersbøll J., Nissen N. I., Hou-Jensen K. A simplified working formulation of non-Hodgkin's lymphomas based on quantifiable histologic criteria. Cancer. 1989 Dec 15;64(12):2532–2540. doi: 10.1002/1097-0142(19891215)64:12<2532::aid-cncr2820641221>3.0.co;2-8. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Swanson S. A., Brooks J. J. Proliferation markers Ki-67 and p105 in soft-tissue lesions. Correlation with DNA flow cytometric characteristics. Am J Pathol. 1990 Dec;137(6):1491–1500. [PMC free article] [PubMed] [Google Scholar]
  35. Vindeløv L. L., Christensen I. J., Jensen G., Nissen N. I. Limits of detection of nuclear DNA abnormalities by flow cytometric DNA analysis. Results obtained by a set of methods for sample-storage, staining and internal standardization. Cytometry. 1983 Mar;3(5):332–339. doi: 10.1002/cyto.990030505. [DOI] [PubMed] [Google Scholar]
  36. Waseem N. H., Lane D. P. Monoclonal antibody analysis of the proliferating cell nuclear antigen (PCNA). Structural conservation and the detection of a nucleolar form. J Cell Sci. 1990 May;96(Pt 1):121–129. doi: 10.1242/jcs.96.1.121. [DOI] [PubMed] [Google Scholar]
  37. Woods A. L., Hall P. A., Shepherd N. A., Hanby A. M., Waseem N. H., Lane D. P., Levison D. A. The assessment of proliferating cell nuclear antigen (PCNA) immunostaining in primary gastrointestinal lymphomas and its relationship to histological grade, S+G2+M phase fraction (flow cytometric analysis) and prognosis. Histopathology. 1991 Jul;19(1):21–27. doi: 10.1111/j.1365-2559.1991.tb00890.x. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. Yu C. C., Fletcher C. D., Newman P. L., Goodlad J. R., Burton J. C., Levison D. A. A comparison of proliferating cell nuclear antigen (PCNA) immunostaining, nucleolar organizer region (AgNOR) staining, and histological grading in gastrointestinal stromal tumours. J Pathol. 1992 Feb;166(2):147–152. doi: 10.1002/path.1711660210. [DOI] [PubMed] [Google Scholar]
  40. Yu C. C., Hall P. A., Fletcher C. D., Camplejohn R. S., Waseem N. H., Lane D. P., Levison D. A. Haemangiopericytomas: the prognostic value of immunohistochemical staining with a monoclonal antibody to proliferating cell nuclear antigen (PCNA). Histopathology. 1991 Jul;19(1):29–33. doi: 10.1111/j.1365-2559.1991.tb00891.x. [DOI] [PubMed] [Google Scholar]

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