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. 1994 Nov;49(11):1152–1156. doi: 10.1136/thx.49.11.1152

Comparison of nucleolar organiser regions and DNA flow cytometry in the evaluation of pleural effusion.

M S Huang 1, M S Tsai 1, J J Hwang 1, T H Wang 1
PMCID: PMC475279  PMID: 7831633

Abstract

BACKGROUND--In conventional cytological diagnosis of pleural effusions the assessment of morphological features plays an important part. However, false negative and false positive results may occur. In this study conventional cytology was compared with flow cytometric DNA analysis and the argyrophil staining technique for nucleolar organiser regions (AgNOR) to characterise benign and malignant effusions. METHODS--Pleural effusions from 71 patients (38 with benign lung disease, 33 with proven adenocarcinoma of lung) were studied by conventional cytology, flow cytometric DNA analysis, and the AgNOR technique. Tumour cell ploidy was determined by flow cytometry. In an attempt to detect the cell proliferative state, flow cytometric S phase fraction and the AgNOR technique were used. The correlations among conventional cytology, flow cytometric DNA ploidy, S phase fraction analysis, and nucleolar organiser regions were investigated. RESULTS--All the 38 benign pleural effusions were diploid. There were 17 (52%) aneuploid and 16 (48%) diploid malignant pleural effusions. Based on these results this type of DNA analysis had a sensitivity of 52% and a specificity of 100%. The mean (SD) numbers of flow cytometric S phase fractions of benign and malignant cases were 5.32 (1.67)% and 12.45 (3.93)% respectively. The mean numbers of S phase fractions of diploid malignant cases were higher than diploid benign cases. In each case the number of AgNORs was counted in 100 cells. The mean number of AgNOR dots per nucleus was 12.57 (3.64) for malignant pleural effusion cells and 3.96 (1.39) for benign pleural effusion cells. The mean number of AgNOR dots was 14.45 (3.36) for aneuploid malignant pleural effusion cells and 10.57 (2.82) for diploid malignant pleural effusion cells. The AgNOR numbers were higher in diploid malignant cells than in diploid benign cells. There was a significant correlation between the S phase fraction determined by flow cytometry and the mean number of AgNORs per nucleus in malignant cases. CONCLUSIONS--Both flow cytometry and the AgNOR methods provide comparable measurements in the diagnosis of pleural effusion. The study also indicates that the AgNOR method, which is rapid and easy to perform, may be a useful adjunct to flow cytometry, S phase fraction analysis and conventional cytology in the routine diagnosis of malignant pleural effusion.

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Selected References

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  1. Ayres J. G., Crocker J. G., Skilbeck N. Q. Differentiation of malignant from normal and reactive mesothelial cells by the argyrophil technique for nucleolar organiser region associated proteins. Thorax. 1988 May;43(5):366–370. doi: 10.1136/thx.43.5.366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barlogie B. Abnormal cellular DNA content as a marker of neoplasia. Eur J Cancer Clin Oncol. 1984 Sep;20(9):1123–1125. doi: 10.1016/0277-5379(84)90119-6. [DOI] [PubMed] [Google Scholar]
  3. Barlogie B., Drewinko B., Schumann J., Göhde W., Dosik G., Latreille J., Johnston D. A., Freireich E. J. Cellular DNA content as a marker of neoplasia in man. Am J Med. 1980 Aug;69(2):195–203. doi: 10.1016/0002-9343(80)90379-4. [DOI] [PubMed] [Google Scholar]
  4. Carbajo S., Orfao A., Vicente-Villardón J. L., Carbajo-Pérez E. Expression of silver-stained nucleolar organizer regions is coupled to cell cycle in rat thymic cells. Cytometry. 1993;14(1):46–52. doi: 10.1002/cyto.990140109. [DOI] [PubMed] [Google Scholar]
  5. Clark G. M., Dressler L. G., Owens M. A., Pounds G., Oldaker T., McGuire W. L. Prediction of relapse or survival in patients with node-negative breast cancer by DNA flow cytometry. N Engl J Med. 1989 Mar 9;320(10):627–633. doi: 10.1056/NEJM198903093201003. [DOI] [PubMed] [Google Scholar]
  6. Crocker J., Ayres J., McGovern J. Nucleolar organiser regions in small cell carcinoma of the bronchus. Thorax. 1987 Dec;42(12):972–975. doi: 10.1136/thx.42.12.972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Crocker J., McGovern J. Nucleolar organiser regions in normal, cirrhotic, and carcinomatous livers. J Clin Pathol. 1988 Oct;41(10):1044–1048. doi: 10.1136/jcp.41.10.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Crocker J., Nar P. Nucleolar organizer regions in lymphomas. J Pathol. 1987 Feb;151(2):111–118. doi: 10.1002/path.1711510203. [DOI] [PubMed] [Google Scholar]
  10. Crocker J., Skilbeck N. Nucleolar organiser region associated proteins in cutaneous melanotic lesions: a quantitative study. J Clin Pathol. 1987 Aug;40(8):885–889. doi: 10.1136/jcp.40.8.885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Evans D. A., Thornthwaite J. T., Ng A. B., Sugarbaker E. V. DNA flow cytometry of pleural effusions. Comparison with pathology for the diagnosis of malignancy. Anal Quant Cytol. 1983 Mar;5(1):19–27. [PubMed] [Google Scholar]
  13. Fallowfield M. E., Dodson A. R., Cook M. G. Nucleolar organizer regions in melanocytic dysplasia and melanoma. Histopathology. 1988 Jul;13(1):95–99. doi: 10.1111/j.1365-2559.1988.tb02007.x. [DOI] [PubMed] [Google Scholar]
  14. Frankfurt O. S., Slocum H. K., Rustum Y. M., Arbuck S. G., Pavelic Z. P., Petrelli N., Huben R. P., Pontes E. J., Greco W. R. Flow cytometric analysis of DNA aneuploidy in primary and metastatic human solid tumors. Cytometry. 1984 Jan;5(1):71–80. doi: 10.1002/cyto.990050111. [DOI] [PubMed] [Google Scholar]
  15. Hall P. A., Crocker J., Watts A., Stansfeld A. G. A comparison of nucleolar organizer region staining and Ki-67 immunostaining in non-Hodgkin's lymphoma. Histopathology. 1988 Apr;12(4):373–381. doi: 10.1111/j.1365-2559.1988.tb01952.x. [DOI] [PubMed] [Google Scholar]
  16. Hanley J. A., McNeil B. J. The meaning and use of the area under a receiver operating characteristic (ROC) curve. Radiology. 1982 Apr;143(1):29–36. doi: 10.1148/radiology.143.1.7063747. [DOI] [PubMed] [Google Scholar]
  17. Hedley D. W., Philips J., Rugg C. A., Taylor I. W. Measurement of cellular DNA content as an adjunct to diagnostic cytology in malignant effusions. Eur J Cancer Clin Oncol. 1984 Jun;20(6):749–752. doi: 10.1016/0277-5379(84)90211-6. [DOI] [PubMed] [Google Scholar]
  18. Henderson A. S., Warburton D., Atwood K. C. Location of ribosomal DNA in the human chromosome complement. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3394–3398. doi: 10.1073/pnas.69.11.3394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Howell W. M., Black D. A. Controlled silver-staining of nucleolus organizer regions with a protective colloidal developer: a 1-step method. Experientia. 1980 Aug 15;36(8):1014–1015. doi: 10.1007/BF01953855. [DOI] [PubMed] [Google Scholar]
  20. Huang M. S., Hwang J. J., Tsai M. S., Wang T. H., Lin M. S., Chong I. W., Tsai S. Y. Application of staining of nucleolar organizer regions in cytological smears of the bronchus. Gaoxiong Yi Xue Ke Xue Za Zhi. 1992 Mar;8(3):136–140. [PubMed] [Google Scholar]
  21. Høstmark J., Vigander T., Skaarland E. Characterization of pleural effusions by flow-cytometric DNA analysis. Eur J Respir Dis. 1985 May;66(5):315–319. [PubMed] [Google Scholar]
  22. Katz R. L., Johnson T. S., Williamson K. D. Comparison of cytologic and acridine-orange flow-cytometric detection of malignant cells in human body cavity fluids. Anal Quant Cytol Histol. 1985 Sep;7(3):227–235. [PubMed] [Google Scholar]
  23. 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]
  24. Nairn E. R., Crocker J., McGovern J. Limited value of AgNOR enumeration in assessment of thyroid neoplasms. J Clin Pathol. 1988 Oct;41(10):1136–1136. doi: 10.1136/jcp.41.10.1136-a. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. O'Hara M. F., Bedrossian C. W., Johnson T. S., Barlogie B. Flow cytometry in cancer diagnosis. Prog Clin Pathol. 1984;9:135–153. [PubMed] [Google Scholar]
  26. Ploton D., Menager M., Adnet J. J. Simultaneous high resolution localization of Ag-NOR proteins and nucleoproteins in interphasic and mitotic nuclei. Histochem J. 1984 Aug;16(8):897–906. doi: 10.1007/BF01002794. [DOI] [PubMed] [Google Scholar]
  27. Schneller J., Eppich E., Greenebaum E., Elequin F., Sherman A., Wersto R., Koss L. G. Flow cytometry and Feulgen cytophotometry in evaluation of effusions. Cancer. 1987 Apr 1;59(7):1307–1313. doi: 10.1002/1097-0142(19870401)59:7<1307::aid-cncr2820590713>3.0.co;2-q. [DOI] [PubMed] [Google Scholar]
  28. Schwarzacher-Robinson T., Kraemer P. M., Cram L. S. Spontaneous in vitro neoplastic evolution of cultured Chinese hamster cells. Nucleolus organizing region activity. Cancer Genet Cytogenet. 1988 Oct 1;35(1):119–128. doi: 10.1016/0165-4608(88)90130-6. [DOI] [PubMed] [Google Scholar]
  29. Smith R., Crocker J. Evaluation of nucleolar organizer region-associated proteins in breast malignancy. Histopathology. 1988 Feb;12(2):113–125. doi: 10.1111/j.1365-2559.1988.tb01923.x. [DOI] [PubMed] [Google Scholar]
  30. Stonesifer K. J., Xiang J. H., Wilkinson E. J., Benson N. A., Braylan R. C. Flow cytometric analysis and cytopathology of body cavity fluids. Acta Cytol. 1987 Mar-Apr;31(2):125–130. [PubMed] [Google Scholar]
  31. Traganos F. Flow cytometry: principles and applications. I. Cancer Invest. 1984;2(2):149–163. doi: 10.3109/07357908409020296. [DOI] [PubMed] [Google Scholar]
  32. Traganos F. Flow cytometry: principles and applications. II. Cancer Invest. 1984;2(3):239–258. doi: 10.3109/07357908409104378. [DOI] [PubMed] [Google Scholar]
  33. Unger K. M., Raber M., Bedrossian C. W., Stein D. A., Barlogie B. Analysis of pleural effusions using automated flow cytometry. Cancer. 1983 Sep 1;52(5):873–877. doi: 10.1002/1097-0142(19830901)52:5<873::aid-cncr2820520522>3.0.co;2-v. [DOI] [PubMed] [Google Scholar]
  34. Weissman G. S., McKinley M. J., Budman D. R., Caccese W. J., Schulman P., Grueneberg D., Bronzo R. L. Flow cytometry. A new technique in the diagnosis of malignant ascites. J Clin Gastroenterol. 1987 Oct;9(5):599–602. doi: 10.1097/00004836-198710000-00025. [DOI] [PubMed] [Google Scholar]

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