Summary
The value of automatic image analysis in the investigation of nucleolus regions (AgNOR) has been examined in tissue sections of 52 malignant and 30 benign breast lesions. Determination of the AgNOR number per cell alone revealed a considerable overlap between benign (range 1.2–3.8) and malignant specimens (range 1.5–16.2). They differed however, highly significantly (P<0.001) in their AgNOR sizes. In benign breast disorders the mean AgNOR area per tumour ranged from 0.22 μm2 to 1.07 μm2 (mean 0.39 μm2), whereas in carcinomas AgNOR sites ranged from 0.05 μm2 to 0.22 μm2 (mean 0.09 μm2). AgNOR counts showed a good correlation with histopathological grade (P<0.05), aneuploidy (P<0.01), proliferation rate as determined by Ki67 immunostaining (P<0.01), as well as oestrogen and progesterone receptor content (P<0.01). Image analysis proved to be advantageous over AgNOR counting alone as it facilitated the standardization of the AgNOR technique itself and thus, significantly improved its diagnostic specifity.
Similar content being viewed by others
Abbreviations
- AgNOR:
-
nucleolar organizer regions demonstrated by silver staining
- Ki67:
-
proliferation marker
References
Barlogie B, Göhde W, Johnston DA (1978) Determination of ploidy of proliferative characteristics of human solid tumours by pulse cytophotometry. Cancer Res 38:3333–3339
Bloom HJG, Richardson WW (1957) Histological grading and prognosis in breast cancer. Br J Cancer 11:359–377
Böcking A, Adler CP, Common HH, Hilgarth M, Granzen B, Auffermann W (1984) Algorithm for a DNA-cytophotometric diagnosis and grading of malignancy. Anal Quant Cytol Histol 6:1–8
Boldy DAR, Crocker J, Ayres JG (1989) Application of the AgNOR method to cell imprints of lymphoid tissue. J Pathol 157:75–79
Crocker J, Nar P (1987) Nucleolar organizer regions in lymphomas. J Pathol 151:111–118
Crocker J, Skilbeck N (1987) Nucleolar organizer region associated proteins in melanotic lesions of the skin. J Clin Pathol 40:885–889
Crocker J, Egan MJ (1988) Correlation between Nor sizes and numbers in non-Hodgkin's-lymphomas. J Pathol 156:233–239
Crocker J, Macartney JC, Smith PJ (1988) Correlation between DNA flow cytometric and nucleolar organizer region data in non-Hodgkin's lymphomas. J Pathol 154:151–156
Crocker J, Boldy DAR, Egan MJ (1989) How should we count AgNORS? Proposals for a standardized approach. J Pathol 158:185–188
Deleener A, Castelain Ph, Preat V, de Gerlache J, Alexandre H, Kirsch-Volders M (1987) Changes in nucleolar transcriptional activity and nuclear DNA content during the first steps of rat hepatocarcinogenesis. Carcinogenesis 8:195–201
Derenzini M, Romagnoli T, Mingazzini P, Marinozzi V (1988) Interphasic nucleolar organizer region distribution as diagnostic parameter to differentiate benign from malignant epithelial tumours of human intestine. Virchows Arch [B] 54:334–340
Derenzini M, Nardi F, Farabegoli F, Ottinetti A, Roncaroli F, Bussolati G (1989a) Distribution of silver-stained interphase nucleolar organizer regions as a parameter to distinguish neoplastic from non-neoplastic reactive cells in human effusions. Acta Cytol 33:491–498
Derenzini M, Pession A, Farabegoli F, Trere D, Badiali M, Dehan P (1989b) Relationship between interphasic nucleolar organizer regions and growth rate in two neuroblastoma cell lines. Am J Pathol 134:925–932
Dressler LG, Seamer LC, Owens MA, Clark GM, McGuire WL (1988) DNA flow cytometry and prognostic factors in 1331 frozen breast cancer specimens. Cancer 61:420–427
Fallenius AG, Auer GU, Carstensen JM (1988) Prognostic significance of DNA measurements in 409 consecutive breast cancer patients. Cancer 62:331–341
Ferraro M, Parantera G (1988) Human NORs show correlation between transcriptional activity, DNase I sensitivity, and hypomethylation. Cytogenet Cell Genet 47:58–61
E.O.R.T.C. (1980) Revision of the standards for the assessment of hormone receptors in human breast cancer: report of the second E.O.R.T.C. workshop 1979. Eur J Cancer 16:1513–1515
Field D, Fitzgerald PH, Sin FYT (1984) Nucleolar silver-staining patterns related to cell cycle phase and cell generation of PHA-stimulated human lymphocytes. Cytobios 41:23–33
Gerdes J, Lelle RJ, Pickartz H et al. (1984) Growth fractions in breast cancers determined in situ with monoclonal antibody Ki67. J Immunol 1222:1710–1717
Giri DD, Nottingham JF, Lawry J, Dundas SAC, Underwood JCE (1989a) Silver-binding nucleolar organizer regions (AgNORs) in benign and malignant breast lesions: correlations with ploidy and growth phase by DNA flow cytometry. J Pathol 157:307–313
Giri DD, Dundas SAC, Sanderson PR, Howat AJ (1989b) Silver binding nucleoli and nucleolar organizer regions in fine needle aspiration cytology of the breast. Acta Cytol 33:173–175
Griffiths AP, Butler CW, Roberts P, Dixon WF, Qiurke P (1989) Silver-stained structures (AgNORs), their dependence on tissue fixation and absence of prognostic relevance in rectal adenocarcinoma. J Pathol 159:121–127
Hall PA, Crocker J, Watts A, Stansfeld AG (1988) A comparison of nucleolar organizer region staining and Ki67 immunostaining in non-Hodgkin's lymphoma. Histopathology 12:373–381
Hedley DW, Friedlander ML, Taylor IW, Rugg CA, Musgrove EA (1983) Method for analysis of cellular DNA content of paraffin-embedded pathological material using flow cytometry. J Histochem Cytochem 31:1333–1335
Hermanek P, Sobin LH (1987) UICC: TNM classification of malignant tumours, 4th edn. Springer, Berlin Heidelberg New York
Howat AJ, Giri DD, Cotton DWK, Slater DN (1989) Nucleolar organizer regions in Spitz nevi and malignant melanomas. Cancer 63:474–478
Jan-Mohamed RM, Armstrong SJ, Crocker J, Leyland MJ, Hulten MA (1989) The relationship between number of interphase NORs and NOR-bearing chromosomes in non-Hodgkin's lymphoma. J Pathol 158:3–7
Jordan G (1987) At the heart of the nucleolus. Nature 329:489–490
Mayhew TM, Cruz L-M (1973) Stereological correction procedures for estimating true volume proportions from biased samples. J Microsc 99:287–299
Meyer JS, Friedman E, McCrate M, Bauer W (1983) Prediction of early course of breast carcinoma by thymidine labeling. Cancer 51:1879–1886
Moot SK, Peters GN, Cheek JH (1987) Tumour hormone receptor status and recurrences in premenopausal node negative breast carcinoma. Cancer 60:382–385
Neumann K, Rüschoff J, Horstmann A, Kalbfleisch, Zwiorek L (1989) Korrelation zwischen immunhistochemisch und biochemisch bestimmten Progesteronrezeptorgehalt sowie Tumourgrading, Tumourausdehnung, Proliferationsrate und Chromatingehalt beim Mammakarzinom. Tumour Diagnostik & Therapie 10:109–114
Ooms ECM, Veldhuizen RW (1989) Argyrophilic proteins of the nucleolar organizer region in bladder-tumours. Virchows Arch[A] 414:365–369
Ploton D, Menager A, Jeannesson P, Himber G, Pigeon F, Adnet JJ (1986) Improvement in the staining and in the visualization of the argyrophilic proteins of the nucleolar organizer region at the optical level. Histochem J 18:5–14
Raymond WA, Leon ASY (1989) Nucleolar organizer regions relate to growth fractions in human breast carcinoma. Hum Pathol 20:741–746
Reeves BR, Casey G, Honeycombe, Smith S (1984) Correlation of differentiation state and silver staining of nucleolar organizers in promyelocytic leukemia cell line HL-60. Cancer Genet Cytogenet 13:159–166
Reiner A, Kolb R, Jakesz R, Schemper M, Spona J (1987) Prognostic significance of steroid hormone receptors and histopathological characterization of human breast cancer. J Cancer Res Clin Oncol 113:285–290
Remmele W, Stegner HE (1987) Vorschlag zur einheitlichen Definierung eines Immunreaktiven Score (IRS) für den immunhistochemischen Östrogenrezeptor Nachweis (ER-ICA) in Mammakarzinomgewebe. Pathologe 8:138–140
Roberts C, Brasch J, Tattersall MH (1987) Ribosomal RNA gene amplification: a selective advantage in tissue culture. Cancer Genet Cytogenet 29:119–127
Rüschoff J, Plate K, Bittinger A, Thomas C (1989) Nucleolar organizer regions (NORs). Basic concepts and practical application in tumour pathology. Pathol Res Pract 185:878–885
Rüschoff J, Plate KH, Contractor H, Kern S, Zimmermann R, Thomas C (1990) Evaluation of nucleolus organizer regions (NORs) by automatic image analysis: a contribution to standardization. J Pathol 161:113–118
Schwarzacher RT, Kraemer PM, Cram LS (1988) Spontaneous in vitro neoplastic evolution of cultured Chinese hamster cells. Nucleolus organizing region activity. Cancer Genet Cytogenet 35:119–128
Smith R, Crocker J (1988) Evaluation of nucleola region-associated proteins in breast malignancy. Histopathology 12:113–125
Smith PJ, Skilbeck N, Harrison A, Crocker J (1988) The effect of a series of fixatives on the AgNOR technique. J Pathol 155:109–112
Stenkvist B, Bengtsson E, Dahlqvist B et al. (1982) Predicting breast cancer recurrence. Cancer 50:2884–2893
World Health Organization (1981) International tumour classification. Breast tumours, 2nd edn. WHO, Geneva
Zatsepina O, Hozak P, Babadjanyan D, Chentsov Y (1988) Quantitative ultrastructural study of nucleolus-organizing regions at some stages of the cell cycle (G0 period, G2 period, mitosis). Biol Cell 62:211–218
Author information
Authors and Affiliations
Additional information
Dedicated to Professor Dr. D. Schmähl on the occasion of his 65th birthday
Supported in part by a grant from the Bundesminister für Forschung und Technologie and by the P. E. Kempes Foundation
Rights and permissions
About this article
Cite this article
Rüschoff, J., Neumann, K., Contractor, H. et al. Assessment of nucleolar organizer regions by automatic image analysis in breast cancer: correlation with DNA content, proliferation rate, receptor status and histopathological grading. J Cancer Res Clin Oncol 116, 480–485 (1990). https://doi.org/10.1007/BF01612998
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01612998