Top

Breast Cancer Research and Treatment

Published in:

01-03-2008 | Clinical Trial

Bone marrow cytokeratin 19 mRNA level is an independent predictor of relapse-free survival in operable breast cancer patients

Authors: Ragne K. Farmen, Oddmund Nordgård, Bjørnar Gilje, Fuad V. Shammas, Jan T. Kvaløy, Satu Oltedal, Reino Heikkilä

Published in: Breast Cancer Research and Treatment | Issue 2/2008

Abstract

Purpose

To study the prognostic significance of elevated cytokeratin 19 (CK19) mRNA levels in the bone marrow (BM) of operable breast cancer patients.

Patients and Methods

From 1998 to 2000, BM was collected from 195 consecutive breast cancer patients immediately prior to surgery and from 34 healthy volunteers. The patients received surgical and adjuvant treatment according to national guidelines at the time. We analyzed the level of CK19 mRNA in the BM samples from patients and normal controls using a real-time RT-PCR assay. The associations with known prognostic factors and the impact of pathological CK19 mRNA levels on patients’ prognosis were investigated.

Results

Using the 99 percentile of the normal control group as a cut-off, 24 (12%) of the 195 patients and 1 (3%) of the 34 volunteers were diagnosed as CK19 mRNA positive. There was no correlation between CK19 BM status and the clinicopathological factors tested. During a median follow-up of 72 months, 7 (29%) of the 24 CK19 mRNA BM positive patients experienced systemic relapse compared to 20 (12%) of the 171 in the CK19 mRNA negative group. The patients with CK19 mRNA-positive BM had significantly shorter systemic recurrence-free survival (P = 0.01) and overall recurrence-free survival (P = 0.005). Multivariate Cox regression showed CK19 mRNA BM status to be an independent predictor of relapse.

Conclusion

Detection of CK19 mRNA in the BM of breast cancer patients by real-time RT-PCR is an independent predictor of relapse-free survival in operable breast cancer patients.

Heimann R, Hellman S (2000) Clinical progression of breast cancer malignant behavior: what to expect and when to expect it. J Clin Oncol 18(3):591–599PubMed

Slade MJ, Singh A, Smith BM, Tripuraneni G, Hall E, Peckitt C et al (2005) Persistence of bone marrow micrometastases in patients receiving adjuvant therapy for breast cancer: results at 4 years. Int J Cancer 114(1):94–100PubMedCrossRef

Harbeck N, Untch M, Pache L, Eiermann W (1994) Tumour cell detection in the bone marrow of breast cancer patients at primary therapy: results of a 3-year median follow-up. Br J Cancer 69(3):566–571PubMed

Diel IJ, Kaufmann M, Costa SD, Holle R, von Minckwitz G, Solomayer EF et al (1996) Micrometastatic breast cancer cells in bone marrow at primary surgery: prognostic value in comparison with nodal status. J Natl Cancer Inst 88(22):1652–1658PubMedCrossRef

Mansi JL, Gogas H, Bliss JM, Gazet JC, Berger U, Coombes RC (1999) Outcome of primary-breast-cancer patients with micrometastases: a long-term follow-up study. Lancet 354(9174):197–202PubMedCrossRef

Braun S, Pantel K, Muller P, Janni W, Hepp F, Kentenich CR et al (2000) Cytokeratin-positive cells in the bone marrow and survival of patients with stage I, II, or III breast cancer. N Engl J Med 342(8):525–533PubMedCrossRef

Solomayer EF, Diel IJ, Salanti G, Hahn M, Gollan C, Schutz F et al (2001) Time independence of the prognostic impact of tumor cell detection in the bone marrow of primary breast cancer patients. Clin Cancer Res 7(12):4102–4108PubMed

Gebauer G, Fehm T, Merkle E, Beck EP, Lang N, Jager W (2001) Epithelial cells in bone marrow of breast cancer patients at time of primary surgery: clinical outcome during long-term follow-up. J Clin Oncol 19(16):3669–3674PubMed

Wiedswang G, Borgen E, Karesen R, Kvalheim G, Nesland JM, Qvist H et al (2003) Detection of isolated tumor cells in bone marrow is an independent prognostic factor in breast cancer. J Clin Oncol 21(18):3469–3478PubMedCrossRef

10.

Pierga JY, Bonneton C, Vincent-Salomon A, de Cremoux P, Nos C, Blin N et al (2004) Clinical significance of immunocytochemical detection of tumor cells using digital microscopy in peripheral blood and bone marrow of breast cancer patients. Clin Cancer Res 10(4):1392–1400PubMedCrossRef

11.

Funke I, Schraut W (1998) Meta-analyses of studies on bone marrow micrometastases: an independent prognostic impact remains to be substantiated. J Clin Oncol 16(2):557–566PubMed

12.

Braun S, Vogl FD, Naume B, Janni W, Osborne MP, Coombes RC et al (2005) A pooled analysis of bone marrow micrometastasis in breast cancer. N Engl J Med 353(8):793–802PubMedCrossRef

13.

Schoenfeld A, Kruger KH, Gomm J, Sinnett HD, Gazet JC, Sacks N et al (1997) The detection of micrometastases in the peripheral blood and bone marrow of patients with breast cancer using immunohistochemistry and reverse transcriptase polymerase chain reaction for keratin 19. Eur J Cancer 33(6):854–861PubMedCrossRef

14.

Ballestrero A, Coviello DA, Garuti A, Nencioni A, Fama A, Rocco I et al (2001) Reverse-transcriptase polymerase chain reaction of the maspin gene in the detection of bone marrow breast carcinoma cell contamination. Cancer 92(8):2030–2035PubMedCrossRef

15.

Fields KK, Elfenbein GJ, Trudeau WL, Perkins JB, Janssen WE, Moscinski LC (1996) Clinical significance of bone marrow metastases as detected using the polymerase chain reaction in patients with breast cancer undergoing high-dose chemotherapy and autologous bone marrow transplantation. J Clin Oncol 14(6):1868–1876PubMed

16.

Slade MJ, Smith BM, Sinnett HD, Cross NC, Coombes RC (1999) Quantitative polymerase chain reaction for the detection of micrometastases in patients with breast cancer. J Clin Oncol 17(3):870–879PubMed

17.

Stathopoulou A, Vlachonikolis I, Mavroudis D, Perraki M, Kouroussis Ch, Apostolaki S et al (2002) Molecular detection of cytokeratin-19-positive cells in the peripheral blood of patients with operable breast cancer: evaluation of their prognostic significance. J Clin Oncol 20(16):3404–3412PubMedCrossRef

18.

Benoy IH, Elst H, Van der Auwera I, Van Laere S, van Dam P, Van Marck E et al (2004) Real-time RT-PCR correlates with immunocytochemistry for the detection of disseminated epithelial cells in bone marrow aspirates of patients with breast cancer. Br J Cancer 91(10):1813–1820PubMedCrossRef

19.

Benoy IH, Elst H, Philips M, Wuyts H, Van Dam P, Scharpe S et al (2006) Real-time RT-PCR detection of disseminated tumour cells in bone marrow has superior prognostic significance in comparison with circulating tumour cells in patients with breast cancer. Br J Cancer 94(5):672–680PubMed

20.

Benoy IH, Elst H, Philips M, Wuyts H, Van Dam P, Scharpe S et al (2006) Prognostic significance of disseminated tumor cells as detected by quantitative real-time reverse-transcriptase polymerase chain reaction in patients with breast cancer. Clin Breast Cancer 7(2):146–152PubMedCrossRef

21.

Shammas FV, Van Eekelen JA, Wee L, Heikkila R, Osland A (1999) Sensitive and quantitative one-step polymerase chain reaction using capillary electrophoresis and fluorescence detection for measuring cytokeratin 19 expression. Scand J Clin Lab Invest 59(8):635–642PubMedCrossRef

22.

Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A et al (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3(7):research0034.1–0034.11

23.

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25(4):402–408PubMedCrossRef

24.

Nordgard O, Kvaloy JT, Farmen RK, Heikkila R (2006) Error propagation in relative real-time reverse transcription polymerase chain reaction quantification models: the balance between accuracy and precision. Anal Biochem 356(2):182–193PubMedCrossRef

25.

Pfaffl MW, Horgan GW, Dempfle L (2002) Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30(9):e36PubMedCrossRef

26.

Grambsch P, Therneau T (1994) Proportional hazards tests and diagnostics based on weighted residuals. Biometrika 81:515–526CrossRef

27.

Cristofanilli M, Budd GT, Ellis MJ, Stopeck A, Matera J, Miller MC et al (2004) Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med 351(8):781–791PubMedCrossRef

28.

Cristofanilli M, Hayes DF, Budd GT, Ellis MJ, Stopeck A, Reuben JM et al (2005) Circulating tumor cells: a novel prognostic factor for newly diagnosed metastatic breast cancer. J Clin Oncol 23(7):1420–1430PubMedCrossRef

29.

Wiedswang G, Borgen E, Schirmer C, Karesen R, Kvalheim G, Nesland JM et al (2006) Comparison of the clinical significance of occult tumor cells in blood and bone marrow in breast cancer. Int J Cancer 118(8):2013–2019PubMedCrossRef

30.

Xenidis N, Perraki M, Kafousi M, Apostolaki S, Bolonaki I, Stathopoulou A et al (2006) Predictive and prognostic value of peripheral blood cytokeratin-19 mRNA-positive cells detected by real-time polymerase chain reaction in node-negative breast cancer patients. J Clin Oncol 24(23):3756–3762PubMedCrossRef

31.

Schlimok G, Funke I, Holzmann B, Gottlinger G, Schmidt G, Hauser H et al (1987) Micrometastatic cancer cells in bone marrow: in vitro detection with anti-cytokeratin and in vivo labeling with anti-17-1A monoclonal antibodies. Proc Natl Acad Sci U S A 84(23):8672–8676PubMedCrossRef

32.

Abd El-Rehim DM, Pinder SE, Paish CE, Bell J, Blamey RW, Robertson JF et al (2004) Expression of luminal and basal cytokeratins in human breast carcinoma. J Pathol 203(2):661–671PubMedCrossRef

33.

Woelfle U, Cloos J, Sauter G, Riethdorf L, Janicke F, van Diest P et al (2003) Molecular signature associated with bone marrow micrometastasis in human breast cancer. Cancer Res 63(18):5679–5684PubMed

34.

Sorlie T, Perou CM, Tibshirani R, Aas T, Geisler S, Johnsen H et al (2001) Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications. Proc Natl Acad Sci U S A 98(19):10869–10874PubMedCrossRef

35.

Chambers AF, Naumov GN, Vantyghem SA, Tuck AB (2000) Molecular biology of breast cancer metastasis. Clinical implications of experimental studies on metastatic inefficiency. Breast Cancer Res 2(6):400–407PubMedCrossRef

36.

Mehes G, Witt A, Kubista E, Ambros PF (2001) Circulating breast cancer cells are frequently apoptotic. Am J Pathol 159(1):17–20PubMed

37.

Mehlen P, Puisieux A (2006) Metastasis: a question of life or death. Nat Rev Cancer 6(6):449–458PubMedCrossRef

Title: Bone marrow cytokeratin 19 mRNA level is an independent predictor of relapse-free survival in operable breast cancer patients
Authors: Ragne K. Farmen
Oddmund Nordgård
Bjørnar Gilje
Fuad V. Shammas
Jan T. Kvaløy
Satu Oltedal
Reino Heikkilä
Publication date: 01-03-2008
Publisher: Springer US
Published in: Breast Cancer Research and Treatment / Issue 2/2008
Print ISSN: 0167-6806
Electronic ISSN: 1573-7217
DOI: https://doi.org/10.1007/s10549-007-9592-x

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Patients and Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 2/2008

Response to neo-adjuvant chemotherapy in women with BRCA1-positive breast cancers

Epidemiology and economic burden of brain metastases among patients with primary breast cancer: results from a US claims data analysis

Breast cancer risk in women with radial scars in benign breast biopsies

Multicenter phase II trial of Genexol-PM, a Cremophor-free, polymeric micelle formulation of paclitaxel, in patients with metastatic breast cancer

Selective ETAR antagonist atrasentan inhibits hypoxia-induced breast cancer cell invasion

HER2 expression and efficacy of preoperative paclitaxel/FAC chemotherapy in breast cancer

Keynote webinar | Spotlight on antibody–drug conjugates in cancer