Breast Cancer

, 10:74 | Cite as

Detection of MUC1 and keratin 19 mRNAs in the bone marrow by quantitative RT-PCR predicts the risk of distant metastasis in breast cancer patients

  • Hiroko Nogi
  • Hiroshi Takeyama
  • Ken Uchida
  • Toshihiko Agata
  • Junko Horiguchi-Yamada
  • Hisashi Yamada
Original Article

Abstract

Background

Early detection of micrometastasis in bone marrow is critical for the prognosis of breast cancer patients. Quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) has been used to detect cancer cells in bone marrow, but its utility as a prognostic factor still remains obscure.

Materials and Methods

Bone marrow samples were aspirated from the anterosuperior iliac spine of 34 patients, immediately after their surgical procedures had been completed. Control samples were also obtained from 10 healthy adult volunteers. The total RNA was extracted from the mononuclear cells, and the expression levels of β-actin, MUC1 and keratin 19 mRNAs were studied by quantitative RT-PCR. Each mRNA level was scored according to the expression level. The sum of these expression scores was defined as the composite expression score, which was employed as the basis of the evaluation.

Results

The mean follow-up period was 45 months. Nine patients developed distant metastases, and one developed local recurrence. The 4-year disease relapse rates were 75% (RR= 19.38; 95% CI: 1.94-193.20), 28% (RR=3.64; 95% CI: 0.43-31.18), and 8.3% for patients with composite expression scores of 5/6, 3/4 and 2, respectively. The difference among the three groups was statistically significant (log-rank test:p = 0.0029), and multivariate analysis also found the composite expression score to be an independent prognostic factor.

Conclusions

Breast cancer patients who show a high composite expression score in bone marrow have a significantly higher risk of recurrence.

Key words

Bone marrow Micrometastases Quantitative RT-PCR MUC1 Keratin 19 

Abbreviations

RT-PCR

Reverse transcriptase-polymerase chain reaction

ER

Estrogen receptor

PR

Progesterone receptor

References

  1. 1).
    Mansi JL, Berger U, Easton D, McDonnell T, Redding WH, Gazet J, Mckinna A, Powles T, Coombes RC: Micrometastases in bone marrow in patients with primary breast cancer: evaluation as an early predictor of bone metastases.Br Med J 295: 1093–1097, 1987.CrossRefGoogle Scholar
  2. 2).
    Mansi JL, Easton D, Berger U, Gazet JC, Ford HT, Dearnaley D, Coombes RC: Bone marrow micrometastases in primary breast cancer: prognostic significance after 6 years’ follow-up.Eur J Cancer 27: 1552–1555, 1991.PubMedGoogle Scholar
  3. 3).
    Cote RJ, Rosen PP, Lesser ML, Old LJ, Osborne MP: Prediction of early relapse in patients with operable breast cancer by detection of occult bone marrow micrometastases.Clin Oncol 9: 1794–1756, 1991.Google Scholar
  4. 4).
    Dearnaley DP, Ormerod MG, Sloane JP: Micrometastases in breast cancer: long-term follow-up of the first patient cohort.Eur J Cancer 27: 236–239, 1991.PubMedGoogle Scholar
  5. 5).
    Harbeck N, Untch M, Pache L, Eiermann W: Tumor 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: 566–571, 1994.PubMedGoogle Scholar
  6. 6).
    Diel IJ, Kaufmann M, Costa SD, Holle R, Minckwitz G: Micrometastatic breast cancer cells in bone marrow at primary surgery: prognostic value in comparison with nodal status.J Natl Cancer Inst 88: 1652–1664, 1996.PubMedCrossRefGoogle Scholar
  7. 7).
    Mansi JL, Gogas H, Bliss JM, Gazet JC, Berger U, Coombes RC: Outcome of primary-breast-cancer patients with micrometastases: a long-term follow-up study.Lancet 354: 197–202, 1999.PubMedCrossRefGoogle Scholar
  8. 8).
    Braun S, Pantel K, Muller P, Janni W, Hepp F, Kentenich CRM, Gastoph S, Wischnik A, Dimpfl T, Kindermann G, Roethmuller G, Schlimok G: Cytokeratin-positive cells in the bone marrow and survival of patients with stage I, II, or III breast cancer.N Engl J Med 342: 525–533, 2000.PubMedCrossRefGoogle Scholar
  9. 9).
    Gerber B, Krause A, Muller H, Richter D, Reimer T, Makovitzky J, Herrning C, Jeschke U, Kundt G, Friese K: Simultaneous immunohistochemical detection of tumor cells in lymph nodes and bone marrow aspirates in breast cancer and its correlation with other prognostic factors.Clin Oncol 19: 960–971, 2001.Google Scholar
  10. 10).
    Roth MS, Antin JH, Bingham El, Ginsberg D: Detection of Philadelphia chromosome positive cells detected by the polymerase chain reaction following bone marrow transplant for chronic myelogenous leukemia.Blood 74: 882–885, 1989.PubMedGoogle Scholar
  11. 11).
    Moss TJ, Sanders DJ: Detection of neuroblastoma cells in blood.Clin Oncol 8: 736–740, 1990.Google Scholar
  12. 12).
    Lindemann F, Schlimok G, Dirschedl P, Witte J, Riethmuller G: Prognostic significance of micrometastatic tumour cells in bone marrow of colorectal cancer patients.Lancet 340: 685–689, 1992.PubMedCrossRefGoogle Scholar
  13. 13).
    Wood DP, Banks ER, Humphreys S, McRoberts JW, Rangnekar VM: Identification of bone marrow micrometastases in patients with prostate cancer.Cancer 74: 2533–2540, 1994.PubMedCrossRefGoogle Scholar
  14. 14).
    Jauch KW, Heiss MM, Gruetzner U, Funke I, Pantel K, Babic R, Eissner HJ, Roethmuller G, Schilberg FW: Prognostic significance of bone marrow micrometastases in patients with gastric cancer.Clin Oncol 14: 1810–1817, 1996.Google Scholar
  15. 15).
    Pantel K, Izbicki J, Passlick B, Angstwurn M, Haussinger K, Thetter O, Riethmuller G: Frequency and prognostic significance of isolated tumour cells in bone marrow of patients with non-small-cell lung cancer without overt metastases.Lancet 347: 649–653, 1996.PubMedCrossRefGoogle Scholar
  16. 16).
    Osborne MP, Asina S, Wong GY, Old LJ, Cote RJ: Immunofluorescent monoclonal antibody detection of breast cancer in bone marrow: sensitivity in a model system.Cancer Res 49: 2510–2513, 1989.PubMedGoogle Scholar
  17. 17).
    Eliss G, Ferguson M, Yamanaka E, Livingston RB, Gown AM: Monoclonal antibodies for detection of occult carcinoma cells in bone marrow of breast cancer patients.Cancer 63: 2509–2514, 1989.CrossRefGoogle Scholar
  18. 18).
    Datta YH, Adams PT, Drobyski WR, Either SP, Terry VH, Roth MS: Sensitive detection of occult breast cancer by the reverse-transcriptase polymerase chain reaction.Clin Oncol 12: 475–482, 1994.Google Scholar
  19. 19).
    Schoenfeld A, Kruger KH, Gomm J, Gazet JC, Sacks N, Bender HG, Luqmani Y, Coombes RC: 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: 854–861, 1997.PubMedCrossRefGoogle Scholar
  20. 20).
    Zippelius A, Kufer P, Honold G, Kollermann MW, Oberneder R, Schlimok G, Reithmuller G, Pantel K: Limitations of reverse-transcriptase polymerase chain reaction analyses for detection of micrometastatic epithelial cancer cells in bone marrow.Clin Oncol 15: 2701–2708, 1997.Google Scholar
  21. 21).
    Memeto T, Vana J, Bedwana RN: Management and survival of female breast cancer.Cancer 45: 2917–2924, 1980.CrossRefGoogle Scholar
  22. 22).
    Rosen PP, Saigo PE, Braun DW, Weathers E, DePalo A: Prediction of recurrence in stage I (T1N0M0) breast carcinoma.Cancer 193: 15–25, 1981.Google Scholar
  23. 23).
    Mcguire W, Abeloff MD, Fisher B: Adjuvant therapy in node negative breast cancer.Breast Cancer Res Treat 13: 97–115, 1989.PubMedCrossRefGoogle Scholar
  24. 24).
    Miura S: The prognostic significance of axillary node metastasis in breast cancer.Jpn J Breast Cancer 10: 452–459, 1995.Google Scholar
  25. 25).
    Redding WH, Coombes RC, Monaghan P, Clink HM, Imrie SF, Dearnaley DP, Ormerod MG, Sloane JP, Gazet JC, Powles TJ, Neville AM: Detection of micrometastases in patients with primary breast cancer.Lancet 2: 1271–1274, 1983.PubMedGoogle Scholar
  26. 26).
    Heid CA, Stevens J, Livak KJ, Williams PM: Real time quantitative PCR.Genome Res 6: 986–994, 1996.PubMedCrossRefGoogle Scholar
  27. 27).
    Gibson UE, Heid CA, Williams PM: A novel method for real time quantitative RT-PCR.Genome Res 6: 995–1001, 1996.PubMedCrossRefGoogle Scholar
  28. 28).
    Miyake Y, Fujiwara Y, Ohue M, Yamamoto H, Sugita Y, Tomita N, Sekimoto M, Shiozaki H, Monden M: Quantification of micrometastases in lymph nodes of colorectal cancer using real-time fluorescence polymerase chain reaction.Int J Oncol 16: 289–293, 2000.PubMedGoogle Scholar
  29. 29).
    Abe M, Kufe D: Characterization of cis-acting elements regulating transcription of the human DF3 breast carcinoma-associated antigen (MUC1) gene.Proc Natl Acad Sci USA 90: 282–286, 1993.PubMedCrossRefGoogle Scholar
  30. 30).
    Brugger W, Buhring HJ, Grunebach F, Vogel W, Muller R, Brummendorf TH, Ziegler BL, Brossat P, Scheding S, Kanz L: Expression of MUC-1 epitopes on normal bone marrow: implications for the detection of micrometastatic tumor cells.Clin Oncol 17: 1535–1544, 1999.Google Scholar
  31. 31).
    Traweek ST, Liu J, Rattifora H: Keratin gene expression in non-epithelial tissues: detection with polymerase chain reaction.Am J Pathol 142: 1111–1118, 1993.PubMedGoogle Scholar

Copyright information

© The Japanese Breast Cancer Society 2003

Authors and Affiliations

  • Hiroko Nogi
    • 1
  • Hiroshi Takeyama
    • 1
  • Ken Uchida
    • 1
  • Toshihiko Agata
    • 2
  • Junko Horiguchi-Yamada
    • 3
  • Hisashi Yamada
    • 4
  1. 1.Departments of Surgery and Environmental MedicineJikei University School of MedicineJapan
  2. 2.Departments of Public Health and Environmental MedicineJikei University School of MedicineJapan
  3. 3.Departments of Oncology, Institute of DNA MedicineJikei University School of MedicineJapan
  4. 4.Departments of Molecular Genetics, Institute of DNA MedicineJikei University School of MedicineJapan

Personalised recommendations