Journal of Cancer Research and Clinical Oncology

, Volume 131, Issue 8, pp 539–546 | Cite as

Isolated tumor cells in the bone marrow (ITC-BM) of breast cancer patients before and after anthracyclin based therapy: influenced by the HER2- and Topoisomerase IIα-status of the primary tumor?

  • C. Schindlbeck
  • W. Janni
  • N. Shabani
  • A. Kornmeier
  • B. Rack
  • D. Rjosk
  • B. Gerber
  • S. Braun
  • H. Sommer
  • K. Friese
Original Paper

Abstract

Purpose: The presence of isolated tumor cells in the bone marrow (ITC-BM) is an independent prognostic factor in all stages of breast cancer. Both the expression/amplification of human epithelial growth factor receptor 2 (HER2) and Topoisomerase IIα (TOP IIa), a key enzyme of DNA replication and main target of anthracyclins, in breast cancer tissue seem to have predictive value regarding the effectiveness of systemic therapies. Methods: To investigate the correlation between these factors and their influence on clinical outcome, tumor tissue of 54 patients who were screened for ITC-BM before and after anthracyclin-based chemotherapy (abCTX) was examined for HER2 and TOP IIa by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Results: By IHC, 31% of the tumors showed positive for HER2 (2+/3+), 14.6% were amplified in FISH. TOP IIa expression (>50%) was found in 13/53 patients (25%), FISH was positive in 5/47 cases (11%). TOP IIa amplification was not seen in cases without HER2 amplification, five of the seven HER2 amplified cases also were amplified for TOP IIa (71% co-amplification). Forty-three patients had adjuvant, seven neo-adjuvant, four palliative abCTX. ITC-BM were present in 24% of patients before and 31% after CTX. Patients with HER2 (IHC, P=0.29) and TOP IIa (FISH, P=0.16) positive tumors tended to stay or become negative in BM status after abCTX and vice versa. After a median follow-up of 44 months (6–127), none of the factors reached significance for overall survival. Yet, patients with HER2 (P=0.16) and TOP IIa (P=0.09) positive tumors showed a trend towards prolonged disease-free survival. Remarkably, none of the TOP IIa FISH-positive patients developed distant metastases (P=0.099) or died (P=0.19) after CTX so far. Conclusions: HER2- and TOP IIa positivity seem to improve the effect of abCTX. The combination of the prognostic value of ITC-BM and the predictive capacity of HER2 and TOP IIa could help to stratify patients for certain therapies. The direct examination of those factors on ITC-BM is the focus of ongoing studies.

Keywords

Breast cancer Isolated tumor cells Bone marrow Minimal residual disease Her2 Topoisomerase Anthracyclin Chemotherapy Prognosis 

Abbreviations

abCTX

Anthracyclin-based chemotherapy

APAAP

Alkaline phosphatase anti-alkaline phosphatase technique

BM

Bone marrow

CEP 17

Centromer enumeration probe chromosome 17

CK

Cytokeratin

CMF

Cyclophosphamide/methotrexat/5-Fluorouracil

DAPI

4′-6′-Diamidino-2′-phenylindole

DDFS

Distant disease-free survival

DFS

Disease-free Survival

DNA

Deoxyribonucleic acid

FISH

Fluorescence in situ hybridization

HER2

Human epithelial growth factor receptor 2

IHC

Immunohistochemistry

ITC

Isolated tumor cells

OS

Overall survival

Top IIa

Topoisomerase IIα

Notes

Acknowledgements

We thank Mrs. S. Schulze for excellent technical assistance. This study contains materials analyzed by Mrs. Anke Kornmeier in preparation of her thesis to achieve the degree of MD at Ludwig-Maximilians University, Medical Faculty, in Munich, Germany.

References

  1. Bartlett J, Mallon E, Cooke T (2003) The clinical evaluation of HER-2 status: which test to use? J Pathol 199:411–417PubMedGoogle Scholar
  2. Braun S, Kentenich C, Janni W, Hepp F, de Waal J, Willgeroth F, Sommer H, Pantel K, (2000a) Lack of effect of adjuvant chemotherapy on the elimination of single dormant tumor cells in bone marrow of high-risk breast cancer patients. J Clin Oncol 18:80–86PubMedGoogle Scholar
  3. Braun S, Pantel K, Muller P, Janni W, Hepp F, Kentenich CR, Gastroph S, Wischnik A, Dimpfl T, Kindermann G, Riethmuller G, Schlimok G (2000b) Cytokeratin-positive cells in the bone marrow and survival of patients with stage I, II, or III breast cancer. N Engl J Med 342:525–533PubMedGoogle Scholar
  4. Braun S, Vogl FD, Schlimok G, Diel IJ, Janni W, Gerber B, Gebauer G, Coombes RC, Pierga J-Y, Naume B, Pantel K (2003) Pooled analysis of prognostic impact of bone marrow micrometastasis: 10-year survival of 4199 breast cancer patients. Presentation, 26th Annual San Antonio Breast Cancer SymposiumGoogle Scholar
  5. Chearskul S, Onreabroi S, Churintrapun M, Semprasert N, Bhothisuwan K (2001) Immunohistochemical study of c-erbB-2 expression in primary breast cancer. Asian Pac J Allergy Immunol 19:197–205PubMedGoogle Scholar
  6. Coleman RE (2003) Current and future status of adjuvant therapy for breast cancer. Cancer 97:880–886PubMedGoogle Scholar
  7. Coon JS, Marcus E, Gupta-Burt S, Seelig S, Jacobson K, Chen S, Renta V, Fronda G, Preisler HD (2002) Amplification and overexpression of topoisomerase IIalpha predict response to anthracycline-based therapy in locally advanced breast cancer. Clin Cancer Res 8:1061–1067PubMedGoogle Scholar
  8. Cortes F, Pastor N, Mateos S, Dominguez I (2003) Roles of DNA topoisomerases in chromosome segregation and mitosis. Mutat Res 543:59–66PubMedGoogle Scholar
  9. Depowski PL, Rosenthal SI, Brien TP, Stylos S, Johnson RL, Ross JS (2000) Topoisomerase IIalpha expression in breast cancer: correlation with outcome variables. Mod Pathol 13:542–547PubMedGoogle Scholar
  10. Di Leo A, Gancberg D, Larsimont D, Tanner M, Jarvinen T, Rouas G, Dolci S, Leroy JY, Paesmans M, Isola J, Piccart MJ (2002) HER-2 amplification and topoisomerase IIalpha gene aberrations as predictive markers in node-positive breast cancer patients randomly treated either with an anthracycline-based therapy or with cyclophosphamide, methotrexate, and 5-fluorouracil. Clin Cancer Res 8:1107–1116PubMedGoogle Scholar
  11. Durbecq V, Di Leo A, Cardoso F, Rouas G, Leroy JY, Piccart M, Larsimont D (2003) Comparison of topoisomerase-IIalpha gene status between primary breast cancer and corresponding distant metastatic sites. Breast Cancer Res Treat 77:199–204PubMedGoogle Scholar
  12. Hellemans P, van Dam PA, Geyskens M, van Oosterom AT, Buytaert P, Van Marck E (1995) Immunohistochemical study of topoisomerase II-alpha expression in primary ductal carcinoma of the breast. J Clin Pathol 48:147–150PubMedGoogle Scholar
  13. Janni W, Hepp F, Rjosk D, Kentenich C, Strobl B, Schindlbeck C, Hantschmann P, Sommer H, Pantel K, Braun S (2001) The fate and prognostic value of occult metastatic cells in the bone marrow of patients with breast carcinoma between primary treatment and recurrence. Cancer 92:46–53PubMedGoogle Scholar
  14. Jarvinen TA, Liu ET (2003) HER-2/neu and topoisomerase IIalpha in breast cancer. Breast Cancer Res Treat 78:299–311PubMedGoogle Scholar
  15. Jarvinen TA, Kononen J, Pelto-Huikko M, Isola J (1996) Expression of topoisomerase IIalpha is associated with rapid cell proliferation, aneuploidy, and c-erbB2 overexpression in breast cancer. Am J Pathol 148:2073–2082PubMedGoogle Scholar
  16. Jarvinen TA, Tanner M, Barlund M, Borg A, Isola J (1999) Characterization of topoisomerase II alpha gene amplification and deletion in breast cancer. Genes Chromosomes Cancer 26:142–150PubMedGoogle Scholar
  17. Jarvinen TA, Tanner M, Rantanen V, Barlund M, Borg A, Grenman S, Isola J (2000) Amplification and deletion of topoisomerase IIalpha associate with ErbB-2 amplification and affect sensitivity to topoisomerase II inhibitor doxorubicin in breast cancer. Am J Pathol 156:839–847PubMedGoogle Scholar
  18. Kakar S, Puangsuvan N, Stevens JM, Serenas R, Mangan G, Sahai S, Mihalov ML (2000) HER-2/neu assessment in breast cancer by immunohistochemistry and fluorescence in situ hybridization: comparison of results and correlation with survival. Mol Diagn 5:199–207PubMedGoogle Scholar
  19. Kaufmann M, von Minckwitz G, Smith R, Valero V, Gianni L, Eiermann W, Howell A, Costa SD, Beuzeboc P, Untch M, Blohmer JU, Sinn HP, Sittek R, Souchon R, Tulusan AH, Volm T, Senn HJ (2003) International expert panel on the use of primary (preoperative) systemic treatment of operable breast cancer: review and recommendations. J Clin Oncol 21:2600–2608PubMedGoogle Scholar
  20. Lebeau A, Deimling D, Kaltz C, Sendelhofert A, Iff A, Luthardt B, Untch M, Lohrs U (2001) Her-2/neu analysis in archival tissue samples of human breast cancer: comparison of immunohistochemistry and fluorescence in situ hybridization. J Clin Oncol 19:354–363PubMedGoogle Scholar
  21. MacGrogan G, Rudolph P, Mascarel II, Mauriac L, Durand M, Avril A, Dilhuydy JM, Robert J, Mathoulin-Pelissier S, Picot V, Floquet A, Sierankowski G, Coindre JM (2003) DNA topoisomerase IIalpha expression and the response toprimary chemotherapy in breast cancer. Br J Cancer 89:666–671PubMedGoogle Scholar
  22. Pantel K, Schlimok G, Braun S, Kutter D, Lindemann F, Schaller G, Funke I, Izbicki JR, Riethmuller G (1993) Differential expression of proliferation-associated molecules in individual micrometastatic carcinoma cells. J Natl Cancer Inst 85:1419–1424PubMedGoogle Scholar
  23. Park K, Kim J, Lim S, Han S (2003) Topoisomerase II-alpha (topoII) and HER2 amplification in breast cancers and response to preoperative doxorubicin chemotherapy. Eur J Cancer 39:631–634PubMedGoogle Scholar
  24. Pauletti G, Dandekar S, Rong H, Ramos L, Peng H, Seshadri R, Slamon DJ (2000) Assessment of methods for tissue-based detection of the HER-2/neu alteration in human breast cancer: a direct comparison of fluorescence in situ hybridization and immunohistochemistry. J Clin Oncol 18:3651–3664PubMedGoogle Scholar
  25. Riou G, Mathieu MC, Barrois M, Le Bihan ML, Ahomadegbe JC, Benard J, Le MG (2001) c-erbB-2 (HER-2/neu) gene amplification is a better indicator of poor prognosis than protein over-expression in operable breast-cancer patients. Int J Cancer 95:266–270PubMedGoogle Scholar
  26. Ross JS, Fletcher JA (1998) The HER-2/neu oncogene in breast cancer: prognostic factor, predictive factor, and target for therapy. Stem Cells 16:413–428PubMedGoogle Scholar
  27. Rudolph P, MacGrogan G, Bonichon F, Frahm SO, de Mascarel I, Trojani M, Durand M, Avril A, Coindre JM, Parwaresch R (1999) Prognostic significance of Ki-67 and topoisomerase IIalpha expression in infiltrating ductal carcinoma of the breast. A multivariate analysis of 863 cases. Breast Cancer Res Treat 55:61–71PubMedGoogle Scholar
  28. Schindlbeck C, Janni W, Shabani N, Rack B, Gerber B, Schmitt M, Harbeck N, Sommer H, Braun S, Friese K (2004) Comparative analysis between the HER2 status in primary breast cancer tissue and the detection of isolated tumor cells in the bone marrow. Breast Cancer Res Treat 87:65–74PubMedGoogle Scholar
  29. Vogel CL, Cobleigh MA, Tripathy D, Gutheil JC, Harris LN, Fehrenbacher L, Slamon DJ, Murphy M, Novotny WF, Burchmore M, Shak S, Stewart SJ, Press M (2002) Efficacy and safety of trastuzumab as a single agent in first-line treatment of HER2-overexpressing metastatic breast cancer. J Clin Oncol 20:719–726PubMedGoogle Scholar
  30. Wang JC (2002) Cellular roles of DNA topoisomerases: a molecular perspective. Nat Rev Mol Cell Biol 3:430–440PubMedGoogle Scholar
  31. Wiedswang G, Borgen E, Karesen R, Qvist H, Janbu J, Kvalheim G, Nesland JM, Naume B (2004) Isolated tumor cells in bone marrow three years after diagnosis in disease-free breast cancer patients predict unfavorable clinical outcome. Clin Cancer Res 10:5342–5348PubMedGoogle Scholar
  32. Yarden Y (2001) Biology of HER2 and its importance in breast cancer. Oncology 61[Suppl 2]:1CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • C. Schindlbeck
    • 1
  • W. Janni
    • 1
  • N. Shabani
    • 1
  • A. Kornmeier
    • 1
  • B. Rack
    • 1
  • D. Rjosk
    • 1
  • B. Gerber
    • 2
  • S. Braun
    • 3
  • H. Sommer
    • 1
  • K. Friese
    • 1
  1. 1.I. FrauenklinikKlinikum der Ludwig-Maximilians-UniversitätMunichGermany
  2. 2.FrauenklinikUniversität RostockRostockGermany
  3. 3.Universitätsklinik für FrauenheilkundeLeopold-Franzens-UniversitätInnsbruckAustria

Personalised recommendations