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Reverse transcriptase/polymerase chain reaction detection of cytokeratin-19 mRNA in bone marrow and blood of breast cancer patients

  • Original Paper
  • Clinical Oncology
  • Published:
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Abstract

A two-step reverse-transcriptase-based polymerase chain reaction (PCR) with nested primer pairs was developed to amplify sensitive and specific cytokeratin-19 (CK-19) mRNA sequences from human breast cancer cells. No CK-19 pseudogene interference was seen. The larger DNA-derived amplification products could be clearly discriminated from mRNA-derived products. The CK-19 message was not amplified from bone marrow or blood of healthy volunteers and patients with haematological malignancies nor from myeloid and lymphoid cell lines. Breast cancer cells were diluted in buffy coat cells up to 10−6 and CK-19 mRNA sought by PCR. The CK-19 message was detected in 14 of 26 blood samples and 14 of 24 marrow samples but in neither of two peripheral blood stem cell samples taken from 35 breast cancer patients. By sequence-analysis control of two of these samples and two cell lines, the amplified DNA fragments were confirmed to be homologous with the CK-19 sequence. The CK-19 message was further sought in matched blood/marrow samples taken from 13 untreated women in the same cohort at the time of diagnosis. In 3 of these, CK-19 RNA was detected in blood and marrow and, in 3 others, only in blood, but never in marrow alone. The results show that CK-19 assay by reverse transcriptase/PCR is a sensitive and specific technique for the detection of cancer cells in bone marrow and blood. It could be helpful in diagnosis and monitoring of metastatic breast cancer and detection of micrometastases. This should be evaluated on larger numbers of patients, with different clinical samples and epithelial malignancies.

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Abbreviations

CK-19 :

cytokeratin 19

RT-PCR :

reverse transcriptase/polymerase chain reaction

PBSC :

peripheral blood stem cell

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Authors and Affiliations

  1. Bone Marrow Transplantation Unit, Department of Oncology/Haematology, University-Hospital Eppendorf, Martinistraße 52, 20246, Hamburg, Germany

    William Krüger, Cordula Krzizanowski, Marcus Stockschläder, Nicolaus Kröger & Axel R. Zander

  2. Department of Gynaecology and Obstetrics, University-Hospital Eppendorf, Hamburg, Germany

    Michael Holweg & Walter Jonat

  3. Department of Clinical Chemistry, University-Hospital Eppendorf, Hamburg, Germany

    Roman Jung

  4. Clinic for Tumorbiology and Medical Oncology, Breisacherstraße 117, 79011, Freiburg i. Br., Germany

    Klaus Mross

Authors
  1. William Krüger
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  2. Cordula Krzizanowski
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  3. Michael Holweg
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  4. Marcus Stockschläder
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  5. Nicolaus Kröger
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  6. Roman Jung
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  7. Klaus Mross
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  8. Walter Jonat
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  9. Axel R. Zander
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Krüger, W., Krzizanowski, C., Holweg, M. et al. Reverse transcriptase/polymerase chain reaction detection of cytokeratin-19 mRNA in bone marrow and blood of breast cancer patients. J Cancer Res Clin Oncol 122, 679–686 (1996). https://doi.org/10.1007/BF01209032

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  • DOI: https://doi.org/10.1007/BF01209032

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