Iberoamerican Journal of Medicine
Iberoamerican Journal of Medicine
Original article

The Correlate between Argyrophilic Nucleolar Organizer Region Stain and Ki-67 Immnochistochemistry in Diagnostic Breast Cancer

Hassan ALgarsh, Hana Abusaida, Fairouz Torjman

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Introduction: This experimental study gauged the value of argyrophilic nucleolar organizer region (AgNOR) staining as a possible technique for the estimation of cell kinetics in conventional histology sections, in benign and malignant breast lesions.
Methods: With a silver staining technique and immunohistochemistry, we associated the numbers of AgNORs and Ki67 scores in 30 breast carcinomas and 10 benign breast lesions.
Results: The mean values of Ag NORs silver stain dots count for normal, benign, grade II and III were 1.28±0.17, 2.83±0.68, 5.23±0.87 and 7.32±0.92, respectively. There was a statistically significant difference (p≤0.001) was noticed between the all individual groups, among the normal and breast lesion as well as among the GII and GIII. Immunohistochemical Results of Ki-67 protein exhibited homogenous golden-brown color in control case and a positive brown granules or diffuse dark brown color in the nuclei of both benign and malignant cases under the 400X magnify examined under the light microscope.
Discussion: AgNOR counts performed on routine formalin-fixed paraffin sections could provide substantial kinetic evidence. Additionally, the difference in AgNOR counts between benign and malignant tumors is such that they may be of diagnostic worth.


Breast cancer; AgNORs; Ki67; Benign; Malignant


1. Cowdry EV. Cancer cells. Philadelphia: Saunders Co; 1955.
2. Thompson CB. Apoptosis in the pathogenesis and treatment of disease. Science. 1995;267(5203):1456-62. doi: 10.1126/science.7878464.
3. Kerr JF, Winterford CM, Harmon BV. Apoptosis. Its significance in cancer and cancer therapy. Cancer. 1994;73(8):2013-26. doi: 10.1002/1097-0142(19940415)73:8<2013::aid-cncr2820730802>3.0.co;2-j.
4. World Health Organization (WHO). Breast cancer: prevention and control. Available from: http://www.who.int/cancer/detection/breastcancer/en/index.html. (accessed Aug 2020)
5. Coleman MP, Quaresma M, Berrino F, Lutz JM, De Angelis R, Capocaccia R, et al. Cancer survival in five continents: a worldwide population-based study (CONCORD). Lancet Oncol. 2008;9(8):730-56. doi: 10.1016/S1470-2045(08)70179-7.
6. Sivridis E, Sims B. Nucleolar organiser regions: new prognostic variable in breast carcinomas. J Clin Pathol. 1990;43(5):390-2. doi: 10.1136/jcp.43.5.390.
7. Smith R, Crocker J. Evaluation of nucleolar organizer region-associated proteins in breast malignancy. Histopathology. 1988;12(2):113-25. doi: 10.1111/j.1365-2559.1988.tb01923.x.
8. Beresford MJ, Wilson GD, Makris A. Measuring proliferation in breast cancer: practicalities and applications. Breast Cancer Res. 2006;8(6):216. doi: 10.1186/bcr1618.
9. Mourad WA, Erkman-Balis B, Livingston S, Shoukri M, Cox CE, Nicosia SV, et al. Argyrophilic nucleolar organizer regions in breast carcinoma. Correlation with DNA flow cytometry, histopathology, and lymph node status. Cancer. 1992;69(7):1739-44. doi: 10.1002/1097-0142(19920401)69:7<1739::aid-cncr2820690715>3.0.co;2-9.
10. Dasgupta A, Ghosh RN, Sarkar R, Laha RN, Ghosh TK, Mukherjee C. Argyrophilic nucleolar organiser regions (AgNORs) in breast lesions. J Indian Med Assoc. 1997;95(9):492-4.
11. Rzymowska J. AgNOR counts and their combination with flow cytometric analyses and clinical parameters as a prognostic indicator in breast carcinoma. Tumori. 1997;83(6):938-42.
12. Ploton D, Menager M, Jeannesson P, Himber G, Pigeon F, Adnet JJ. Improvement in the staining and in the visualization of the argyrophilic proteins of the nucleolar organizer region at the optical level. Histochem J. 1986;18(1):5-14. doi: 10.1007/BF01676192.
13. Fallowfield ME, Dodson AR, Cook MG. Nucleolar organizer regions in melanocytic dysplasia and melanoma. Histopathology. 1988;13(1):95-9. doi: 10.1111/j.1365-2559.1988.tb02007.x.
14. Darnell J, Lodish H, Baltimore D. Molecular cell biology. New York: Scientific American Books: 1986.
15. Liao GS, Apaya MK, Shyur LF. Herbal medicine and acupuncture for breast cancer palliative care and adjuvant therapy. Evid Based Complement Alternat Med. 2013;2013:437948. doi: 10.1155/2013/437948.
16. Kuru B. Prognostic significance of total number of nodes removed, negative nodes removed, and ratio of positive nodes to removed nodes in node positive breast carcinoma. Eur J Surg Oncol. 2006;32(10):1082-8. doi: 10.1016/j.ejso.2006.06.005.
17. Shi WB, Yang LJ, Hu X, Zhou J, Zhang Q, Shao ZM. Clinico-pathological features and prognosis of invasive micropapillary carcinoma compared to invasive ductal carcinoma: a population-based study from China. PLoS One. 2014;9(6):e101390. doi: 10.1371/journal.pone.0101390.
18. Sofi GN, Sofi JN, Nadeem R, Shiekh RY, Khan FA, Sofi AA, et al. Estrogen receptor and progesterone receptor status in breast cancer in relation to age, histological grade, size of lesion and lymph node involvement. Asian Pac J Cancer Prev. 2012;13(10):5047-52. doi: 10.7314/apjcp.2012.13.10.5047.
19. Hüsemann Y, Geigl JB, Schubert F, Musiani P, Meyer M, Burghart E, et al. Systemic spread is an early step in breast cancer. Cancer Cell. 2008;13(1):58-68. doi: 10.1016/j.ccr.2007.12.003.
20. Zhao X, Malhotra GK, Lele SM, Lele MS, West WW, Eudy JD, et al. Telomerase-immortalized human mammary stem/progenitor cells with ability to self-renew and differentiate. Proc Natl Acad Sci U S A. 2010;107(32):14146-51. doi: 10.1073/pnas.1009030107.
21. EL-Fiky SH, Yahia MA, AL Sedfy AS, AL-Qadasi SA. Immunohistochemical Expression of Fn14 in Invasive Ductal Carcinoma (IDC) and its correlation with clinical and histopathological parameters of human breast cancer. Int Res J Medical Sci. 2015;3(2):7-15.
22. Bhatt J, Patel T, Sarvaiya S, Modha D, Gajjar M. Silver stained nucleolar organizer region count (AgNOR count)–very useful tool in breast lesions. Natl J Med Res. 2013;3:280-2.
23. Dowsett M, Nielsen TO, A'Hern R, Bartlett J, Coombes RC, Cuzick J, et al. Assessment of Ki67 in breast cancer: recommendations from the International Ki67 in Breast Cancer working group. J Natl Cancer Inst. 2011;103(22):1656-64. doi: 10.1093/jnci/djr393.
24. Kill IR. Localisation of the Ki-67 antigen within the nucleolus. Evidence for a fibrillarin-deficient region of the dense fibrillar component. J Cell Sci. 1996;109 ( Pt 6):1253-63.
25. Verheijen R, Kuijpers HJ, van Driel R, Beck JL, van Dierendonck JH, Brakenhoff GJ, et al. Ki-67 detects a nuclear matrix-associated proliferation-related antigen. II. Localization in mitotic cells and association with chromosomes. J Cell Sci. 1989;92 (Pt 4):531-40.
26. Gerdes J, Li L, Schlueter C, Duchrow M, Wohlenberg C, Gerlach Cet al. Immunobiochemical and molecular biologic characterization of the cell proliferation-associated nuclear antigen that is defined by monoclonal antibody Ki-67. Am J Pathol. 1991;138(4):867-73.
27. Koike T, Ohtsuki K. Purification and characterization of a 400-kDa nonhistone chromatin protein that serves as an effective phosphate acceptor for casein kinase II from Ehrlich ascites tumor cells. J Biochem. 1988;103(6):928-37. doi: 10.1093/oxfordjournals.jbchem.a122389.
28. Cazzaniga M, Severi G, Casadio C, Chiapparini L, Veronesi U, Decensi A. Atypia and Ki-67 expression from ductal lavage in women at different risk for breast cancer. Cancer Epidemiol Biomarkers Prev. 2006;15(7):1311-5. doi: 10.1158/1055-9965.EPI-05-0810.
29. Straver ME, Glas AM, Hannemann J, Wesseling J, van de Vijver MJ, Rutgers EJ, et al. The 70-gene signature as a response predictor for neoadjuvant chemotherapy in breast cancer. Breast Cancer Res Treat. 2010;119(3):551-8. doi: 10.1007/s10549-009-0333-1.
30. Ansari HA, Mehdi G, Maheshwari V, Siddiqui SA. Evaluation of AgNOR scores in aspiration cytology smears of breast tumors. J Cytol. 2008;25(3):100-4.
31. Pich A, Chiusa L, Margaria E. Prognostic relevance of AgNORs in tumor pathology. Micron. 2000;31(2):133-41. doi: 10.1016/s0968-4328(99)00070-0.
32. Ceccarelli C, Trerè D, Santini D, Taffurelli M, Chieco P, Derenzini M. AgNORs in breast tumours. Micron. 2000;31(2):143-9. doi: 10.1016/s0968-4328(99)00071-2.
33. Howell WM. Selective staining of nucleolus organizer regions (NORs). In: Busch H, Rothblum L, editors. The cell nucleus. New York: Academic Press; 1982:89-143.
34. Derenzini M, Ceccarelli C, Santini D, Taffurelli M, Treré D. The prognostic value of the AgNOR parameter in human breast cancer depends on the pRb and p53 status. J Clin Pathol. 2004;57(7):755-61. doi: 10.1136/jcp.2003.015917.
35. Shimahara H, Hirano T, Ohya K, Matsuta S, Seeram SS, Tate S. Nucleosome structural changes induced by binding of non-histone chromosomal proteins HMGN1 and HMGN2. FEBS Open Bio. 2013 28;3:184-91. doi: 10.1016/j.fob.2013.03.002.
36. Tuominen VJ, Ruotoistenmäki S, Viitanen A, Jumppanen M, Isola J. ImmunoRatio: a publicly available web application for quantitative image analysis of estrogen receptor (ER), progesterone receptor (PR), and Ki-67. Breast Cancer Res. 2010;12(4):R56. doi: 10.1186/bcr2615.
37. Varga Z, Cassoly E, Li Q, Oehlschlegel C, Tapia C, Lehr HA, et al. Standardization for Ki-67 assessment in moderately differentiated breast cancer. A retrospective analysis of the SAKK 28/12 study. PLoS One. 2015;10(4):e0123435. doi: 10.1371/journal.pone.0123435.
38. Rodman TC, Tahiliani S. The Feulgen banded karyotype of the mouse: analysis of the mechanisms of banding. Chromosoma. 1973;42(1):37-56. doi: 10.1007/BF00326329.
39. Biesterfeld S, Beckers S, Del Carmen Villa Cadenas M, Schramm M. Feulgen staining remains the gold standard for precise DNA image cytometry. Anticancer Res. 2011;31(1):53-8.
40. Dawson AE, Norton JA, Weinberg DS. Comparative assessment of proliferation and DNA content in breast carcinoma by image analysis and flow cytometry. Am J Pathol. 1990;136(5):1115-24.

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