Molecular Diagnosis & Therapy

, Volume 10, Issue 1, pp 41–47 | Cite as

Detection of Circulating Tumor Cells in Peripheral Blood of Breast Cancer Patients During or After Therapy Using a Multigene Real-Time RT-PCR Assay

  • Barbara K. Zehentner
  • Heather Secrist
  • Dawn C. Hayes
  • Xinqun Zhang
  • Richard C. Ostenson
  • Steven Loop
  • Gary Goodman
  • Raymond L. Houghton
  • David H. Persing
Original Research Article


Aim: To evaluate the utility of a multigene real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay to detect circulating tumor cells in peripheral blood specimens of breast cancer patients during or after treatment.

Method: Using this assay, peripheral blood samples were analyzed for expression levels of mammaglobin and three complementary transcribed breast cancer-specific genes: B305D, γ-aminobutyrate type A receptor π subunit (GABA π; GABRP), and B726P. We examined 172 blood specimens from 82 breast cancer patients during or after therapy for the presence of circulating tumor cells using the multigene real-time RT-PCR assay.

Results: In 63.4% of the blood samples, a positive signal for mammaglobin and/or three breast cancer-associated gene transcripts was detected. Of breast cancer patients, 75.6% had at least one positive blood sample. Blood specimens from 51 of 53 healthy female volunteers tested negative in the assay whereas two samples had a low expression signal. In addition, three patients were monitored for more than a year during their adjuvant therapy treatment.

Conclusion: This assay could be a valuable tool for monitoring breast cancer patients during and after therapy.


  1. 1.
    Zehentner BK. Detection of disseminated tumor cells: strategies and diagnostic implications. Expert Rev Mol Diagn 2002; 2(1): 41–8PubMedCrossRefGoogle Scholar
  2. 2.
    Ghossein RA, Rosai J. Polymerase chain reaction in the detection of micrometastases and circulating tumor cells. Cancer 1996; 78(1): 10–6PubMedCrossRefGoogle Scholar
  3. 3.
    Gerhard M, Juhl H, Kalthoff H, et al. Specific detection of carcinoembryonic antigen-expressing tumor cells in bone marrow aspirates by polymerase chain reaction. J Clin Oncol 1994; 12(4): 725–9PubMedGoogle Scholar
  4. 4.
    Slade MJ, Smith BM, Sinnett HD, et al. Quantitative polymerase chain reaction for the detection of micrometastases in patients with breast cancer. J Clin Oncol 1999; 17(3): 870–9PubMedGoogle Scholar
  5. 5.
    de Cremoux P, Extra JM, Deni MG, et al. Detection of MUC1-expressing mammary carcinoma cells in the peripheral blood of breast cancer patients by real-time polymerase chain reaction. Clin Cancer Res 2000; 6(8): 3117–22PubMedGoogle Scholar
  6. 6.
    Zach O, Kasparu H, Krieger O, et al. Detection of circulating mammary carcinoma cells in the peripheral blood of breast cancer patients via a nested reverse transcriptase polymerase chain reaction for mammaglobin mRNA. J Clin Oncol 1999; 17(7): 2015–9PubMedGoogle Scholar
  7. 7.
    Corradini P, Voena C, Astolfi M, et al. Maspin and mammaglobin genes are specific markers for RT-PCR detection of minimal residual disease in patients with breast cancer. Ann Oncol 2001; 12(12): 1693–8PubMedCrossRefGoogle Scholar
  8. 8.
    Watson MA, Fleming TP. Mammaglobin, a mammary-specific member of the uteroglobin gene family, is overexpressed in human breast cancer. Cancer Res 1996; 56(4): 860–5PubMedGoogle Scholar
  9. 9.
    Klug J, Beier HM, Bernard A, et al. Uteroglobin/Clara cell 10-kDa family of proteins: nomenclature committee report. Ann N Y Acad Sci 2000; 923: 348–54PubMedCrossRefGoogle Scholar
  10. 10.
    Ni J, Kalff-Suske M, Gentz R, et al. All human genes of the uteroglobin family are localized on chromosome 1 1q12.2 and form a dense cluster. Ann N Y Acad Sci 2000; 923: 25–42PubMedCrossRefGoogle Scholar
  11. 11.
    Min CJ, Tafra L, Verbanac KM. Identification of superior markers for polymerase chain reaction detection of breast cancer metastases in sentinel lymph nodes. Cancer Res 1998; 58(20): 4581–4PubMedGoogle Scholar
  12. 12.
    Colpitts TL, Billing-Medel P, Friedman P, et al. Mammaglobin is found in breast tissue as a complex with BU101. Biochemistry 2001; 40(37): 11048–59PubMedCrossRefGoogle Scholar
  13. 13.
    Watson MA, Dintzis S, Darrow CM, et al. Mammaglobin expression in primary, metastatic, and occult breast cancer. Cancer Res 1999; 59: 3028–31PubMedGoogle Scholar
  14. 14.
    Fleming TP, Watson MA. Mammaglobin, a breast-specific gene, and its utility as a marker for breast cancer. Ann N Y Acad Sci 2000; 923: 78–89PubMedCrossRefGoogle Scholar
  15. 15.
    Leygue E, Snell L, Dotzlaw H, et al. Mammaglobin, a potential marker of breast cancer nodal metastasis. J Pathol 1999; 189: 28–33PubMedCrossRefGoogle Scholar
  16. 16.
    Houghton RL, Dillon DC, Molesh DA, et al. Transcriptional complementarity in breast cancer: application to detection of circulating tumor cells. Mol Diagn 2001; 6(2): 79–91PubMedCrossRefGoogle Scholar
  17. 17.
    Jiang Y, Harlocker SL, Molesh DA, et al. Discovery of differentially expressed genes in human breast cancer using subtracted cDNA libraries and cDNA microarrays. Oncogene 2002; 21(14): 2270–82PubMedCrossRefGoogle Scholar
  18. 18.
    Hedblom E, Kirkness EF. A novel class of GABAA receptor subunit in tissues of the reproductive system. J Biol Chem 1997; 272: 15346–50PubMedCrossRefGoogle Scholar
  19. 19.
    Jager D, Stocken E, Gure AO, et al. Identification of a tissue-specific putative transcription factor in breast tissue by serological screening of a breast cancer library. Cancer Res 2001; 61: 2055–61PubMedGoogle Scholar
  20. 20.
    Zehentner BK, Dillon DC, Jiang Y, et al. Application of a multigene reverse transcription-PCR assay for detection of mammaglobin and complementary transcribed genes in breast cancer lymph nodes. Clin Chem 2002; 48(8): 1225–31PubMedGoogle Scholar
  21. 21.
    Zehentner BK, Persing DH, Deme A, et al. Mammaglobin as a novel breast cancer biomarker: multigene reverse transcription-PCR assay and sandwich ELISA. Clin Chem 2004; 50(11): 2069–76PubMedCrossRefGoogle Scholar
  22. 22.
    Weigelt B, Bosma AJ, Hart AA, et al. Marker genes for circulating tumour cells predict survival in metastasized breast cancer patients. Br J Cancer 2003; 88(7): 1091–4PubMedCrossRefGoogle Scholar
  23. 23.
    Cristofanilli M, Budd GT, Ellis MJ, et al. Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med 2004; 351(8): 781–91PubMedCrossRefGoogle Scholar

Copyright information

© Adis Data Information BV 2006

Authors and Affiliations

  • Barbara K. Zehentner
    • 1
  • Heather Secrist
    • 2
  • Dawn C. Hayes
    • 2
  • Xinqun Zhang
    • 2
  • Richard C. Ostenson
    • 3
  • Steven Loop
    • 3
  • Gary Goodman
    • 4
  • Raymond L. Houghton
    • 5
  • David H. Persing
    • 2
  1. 1.HematoLogics Inc.SeattleUSA
  2. 2.Corixa CorporationSeattleUSA
  3. 3.Good Samaritan Cancer CenterPuyallupUSA
  4. 4.Swedish Medical CenterSeattleUSA
  5. 5.InBios International Inc.SeattleUSA
  6. 6.GSK-Bio-Seattle (formerly Corixa Corporation)SeattleUSA

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