Breast Cancer Research and Treatment

, Volume 38, Issue 3, pp 265–275 | Cite as

S-phase fraction identifies high-risk subgroups among DNA-diploid breast cancers

  • H. Romero
  • J. Schneider
  • J. Burgos
  • J. Bilbao
  • F. J. Rodriguez-Escudero


The prognostic value of DNA content measured by means of flow cytometry was analyzed in formalin-fixed, paraffin-embedded samples from 231 breast cancer patients treated between 1984 and 1988, with a mean follow-up period of 55 months. We followed the guidelines of a Consensus Meeting held on this issue in Maine, USA, in 1992. DNA-diploid and -aneuploid tumors were evaluated separately for the fraction of cells in S-phase (SPF) contained in them, this being divided into three groups (‘high’, ‘intermediate’, and ‘low’), defined by the 25th and 75th centile of the SPF-distribution corresponding to either DNA-diploid or DNA-aneuploid tumors.

Unequivocally readable histograms were obtained from 174 samples (75.3%).

A high SPF in diploid tumors was significantly associated with a higher recurrence rate (p = 0.015), a shorter disease-free survival (p = 0.014), advanced (IIIB) clinical stage (p = 0.034), and almost significantly with total survival (p = 0.055).

In a multivariate Cox regression analysis, a high SPF in diploid tumors retained its independent prognostic power, being significantly associated with a shorter disease-free survival (p = 0.00049) and total survival (p = 0.0077). It also allowed to identify a subgroup with an ominous prognosis among patients ≤ 50 years of age with early stage tumors.

Our results fully validate the recommendations of the 1992 Maine Consensus Meeting.

Key words

cancer breast prognosis DNA content flow cytometry 


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  1. 1.
    Fisher B, Ravdin RG, Ausman RK, Slack NH, Moore GE, Noer RJ: Surgical adjuvant chemotherapy in cancer of the breast: results of a decade of cooperative investigation. Ann Surg 168: 337–356, 1968PubMedGoogle Scholar
  2. 2.
    Frierson HF Jr: Ploidy analysis and S-phase fraction determination by flow cytometry of invasive adenocarcinomas of the breast. Am J Surg Pathol 15: 358–367, 1991PubMedGoogle Scholar
  3. 3.
    Page DL: Prognosis and breast cancer. Recognition of lethal and favorable prognostic types. Am J Surg Pathol 15: 334–349, 1991PubMedGoogle Scholar
  4. 4.
    Shankey TV, Ravinovitch PS, Bruce B, Bauer K, Cox C, Duque RE, Hedley DW, Mayall BH, Wheeless L: Guidelines for the implementation of clinical DNA cytometry. Breast Cancer Res Treat 28: 61–68, 1993Google Scholar
  5. 5.
    Hedley DW, Clark GM, Cornelisse C, Killander D, Kute T, Merkel D: Consensus review of the clinical utility of DNA cytometry in carcinoma of the breast. Breast Cancer Res Treat 28: 55–59, 1993PubMedGoogle Scholar
  6. 6.
    Hedley DW, Friedlander MI, Taylor IW, Rugg CA, Musgrove EA: Method for analysis of cellular DNA content of paraffin-embedded pathological material using flow cytometry. J Histochem Cytochem 31: 1333–1335, 1983PubMedGoogle Scholar
  7. 7.
    Vindelov LL, Christensen IJ, Nissen NI: A detergent-trypsin method for the preparation of nuclei for flow cytometric DNA analysis. Cytometry 3: 323–327, 1983PubMedGoogle Scholar
  8. 8.
    Kaplan EL, Meier P: Nonparametric estimation from incomplete observations. J Am Stat Assoc 53: 457–481, 1958Google Scholar
  9. 9.
    Mantel N: Evaluation of survival data and two new rank order statistics arising in its consideration. Cancer Chemother Rep 50: 163–170, 1966PubMedGoogle Scholar
  10. 10.
    Cox DR: Regression models and life tables. J R Stat Soc (B) 34: 187–220, 1972Google Scholar
  11. 11.
    UICC International Union Against Cancer. T N M classification of malignant tumors, Third Edition, Geneva, 1988Google Scholar
  12. 12.
    Kallioniemi O-P: Comparison of fresh and paraffin-embedded tissue as starting material for DNA flow cytometry and evaluation of intratumor heterogeneity. Cytometry 9: 164–169, 1988PubMedGoogle Scholar
  13. 13.
    Koss LG, Bogdan C, Herz F, Wersto RP: Flow cytometric measurements of DNA and other cell components in human tumors: a critical appraisal. Human Pathol 20: 528–548, 1989Google Scholar
  14. 14.
    Frierson HF Jr: The need for improvement in flow cytometric analysis of ploidy and S-phase fraction. Am J Clin Pathol 95: 439–441, 1991PubMedGoogle Scholar
  15. 15.
    Fisher B, Gunduz N, Costantino J, Fisher ER, Redmond C, Mamounas EP, Siderits R: DNA flow cytometric analysis of primary operable breast cancer. Cancer 68: 465–475, 1991Google Scholar
  16. 16.
    Clark GM, Dressler LG, Owens MA, Pounds G, Oldaker T, McGuire WL: Prediction of relapse or survival in patients with node-negative breast cancer by DNA flow cytometry. N Engl J Med 320: 627–633, 1989PubMedGoogle Scholar
  17. 17.
    Sigurdsson H, Baldetorp B, Borg A, Dalberg M, Ferno M, Killander D, Olsson H: Indicators of prognosis in node-negative breast cancer. N Engl J Med 322: 1045–1053, 1990PubMedGoogle Scholar
  18. 18.
    Weaver DL, Bagwell CB, Hitchcox SA, Whetstone SD, Baker DR, Herbert DJ, Jones MA: Improved flow cytometric determination of proliferative activity (S-phase fraction) from paraffin-embedded tissue. Am J Clin Pathol 94: 576–584, 1990PubMedGoogle Scholar
  19. 19.
    Dowle CS, Owainati A, Robins A, Burns K, Ellis IO, Elston CW, Blamey RW: Prognostic significance of the DNA content of human breast cancer. Br J Surg 74: 133–136, 1987PubMedGoogle Scholar
  20. 20.
    Dressler LG, Seamer LC, Owens MA, Clark GM, McGuire WL: DNA flow cytometry and prognostic factors in 1331 frozen breast cancer specimens. Cancer 61: 420–427, 1988PubMedGoogle Scholar
  21. 21.
    Furh JE, Frye A, Kattine AA, Van Metter S: Flow cytometric determination of breast tumor heterogeneity. Cancer 67: 1401–1405, 1991PubMedGoogle Scholar
  22. 22.
    Joensuu H, Kallioniemi O-P: Different opinions on classification of DNA histograms produced from paraffin-embedded tissue. Cytometry 10: 711–717, 1989PubMedGoogle Scholar
  23. 23.
    Kallioniemi O-P, Joensuu H, Klemi P, Koivula T: Inter-laboratory comparison of DNA flow cytometric results from paraffin-embedded breast carcinomas. Breast Cancer Res Treat 17: 59–61, 1990PubMedGoogle Scholar
  24. 24.
    Koss LG, Wersto RP, Simmons DA, Deitch D, Hertz F, Freed S: Predictive value of DNA measurements in bladder washings. Comparison of flow cytometry, image cytophotometry, and cytology in patients with a past history of urothelial tumors. Cancer 64: 916–924, 1989PubMedGoogle Scholar
  25. 25.
    Baldetorp B, Ferno M, Fallenius A, Fallenius-Vecchi G, Idvall I, Olsson H, Sigurdsson H, Akerman M, Killander D: Image cytometric DNA analysis in human breast cancer analysis may add prognostic information in diploid cases with low S-phase fraction by flow cytometry. Cytometry 13: 577–585, 1992PubMedGoogle Scholar
  26. 26.
    Hedley DW, Rugg CA, Gelber RD: Association of DNA index and S-phase fraction with prognosis of node positive early breast cancer. Cancer Res 47: 4729–4735, 1987PubMedGoogle Scholar
  27. 27.
    Lewis WE: Prognostic significance of flow cytometric DNA analysis in node-negative breast cancer patients. Cancer 65: 2315–2320, 1990PubMedGoogle Scholar
  28. 28.
    Thorud E, Fossa SD, Vaage S, Kaalhus O, Knudsen OS, Bormer O, Shoaib MC: Primary breast cancer. Flow cytometric DNA pattern in relation to clinical and histopathologic characteristics. Cancer 57: 808–811, 1986PubMedGoogle Scholar
  29. 29.
    Beerman H, Kluin M, Van de Velde CJH, Hermans J, Cornelisse CJ: DNA flow cytometry in the prognosis of node-negative breast cancer. N Engl J Med 321: 473–474, 1989PubMedGoogle Scholar
  30. 30.
    Keyhani-Rofagha S, O'Toole RV, Farrar WB, Sickle-Santanello B, DeCenzo J, Young D: Is DNA ploidy an independent prognostic indicator in infiltrative node-negative breast adenocarcinoma? Cancer 65: 1577–1582, 1990PubMedGoogle Scholar
  31. 31.
    Muss HB, Kute TE, Case LD, Smith LR, Booher C, Long R, Kammire L, Gregory B, Brockschmidt JK: The relation of flow cytometry to clinical and biologic characteristics in women with node negative primary breast cancer. Cancer 64: 1894–1900, 1989PubMedGoogle Scholar
  32. 32.
    Noguchi M, Taniya T, Ohta N, Koyasaki N, Miyasaki I, Mizukami Y: Lymph node metastases versus DNA ploidy as prognostic factors for invasive ductal carcinoma of the breast. Breast Cancer Res Treat 19: 23–31, 1991PubMedGoogle Scholar
  33. 33.
    Sharma S, Mishra MC, Kapur BML, Verma K, Nath I: The prognostic significance of ploidy analysis in operable breast cancer. Cancer 68: 2612–2616, 1991PubMedGoogle Scholar
  34. 34.
    Hedley DW: Developments in the use of flow cytometry as a guide to the prognosis of cancer. Diagn Oncol 1: 2–4, 1991Google Scholar
  35. 35.
    O'Reilly SM, Camplejohn RS, Barnes DM, Millis RR, Rubens RD, Richards MA: Node-negative breast cancer: Prognostic subgroups defined by tumor size and flow cytometry. J Clin Oncol 8: 2040–2046, 1990PubMedGoogle Scholar
  36. 36.
    Clark GM, Mathieu M-C, Owens MA, Dressler LG, Eudey L, Tormey DC, Osborne CK, Gilchrist KW, Mansour EG, Abeloff MD, McGuire WL: Prognostic significance of Sphase fraction in good risk, node-negative breast cancer patients. J Clin Oncol 10: 428–432, 1992PubMedGoogle Scholar
  37. 37.
    Ewers S-B, Attewell R, Baldetorp B, Borg A, Langstrom E, Killander D: Prognostic potential of flow cytometric S-phase and ploidy prospectively determined in primary breast carcinomas. Breast Cancer Res Treat 20: 93–108, 1991Google Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • H. Romero
    • 1
    • 2
  • J. Schneider
    • 1
  • J. Burgos
    • 3
  • J. Bilbao
    • 4
  • F. J. Rodriguez-Escudero
    • 1
  1. 1.Department of Obstetrics and Gynecology, Hospital de CrucesUniversidad del País VascoBaracaldo (Bilbao)Spain
  2. 2.Department of SurgeryUniversidad del CaucaPopayanColombia
  3. 3.Department of Pathology, Hospital de CrucesUniversidad del País VascoBilbaoSpain
  4. 4.Department of Epidemiology and BiostatisticsUniversidad del País VascoBilbaoSpain

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