gms | German Medical Science

27. Deutscher Krebskongress

Deutsche Krebsgesellschaft e. V.

22. - 26.03.2006, Berlin

Immunomagnetic enrichment and molecular detectionof tumor cells in peripheral blood of patients with primary breast cancer

Meeting Abstract

  • corresponding author presenting/speaker Nikos Fersis - Universitäts-Frauenklinik Heidelberg, Deutschland
  • Verena Deckwart - Universitäts-Frauenklinik Heidelberg
  • Alexandra Leitz - Universitäts-Frauenklinik Heidelberg
  • Marion Weber - Universitäts-Frauenklinik Heidelberg
  • Joachim Rom - Universitäts-Frauenklinik Heidelberg
  • Veit Zieglschmid - Adnagen AG, Langenhagen
  • Oliver Böcher - Adnagen AG, Langenhagen
  • Gunther Bastert - Klinik Bad Trissl
  • Christoph Sohn - Universitäts-Frauenklinik Heidelberg
  • Sepp Kaul - Universitäts-Frauenklinik Heidelberg

27. Deutscher Krebskongress. Berlin, 22.-26.03.2006. Düsseldorf, Köln: German Medical Science; 2006. DocOP419

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Veröffentlicht: 20. März 2006

© 2006 Fersis et al.
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Background: The purpose of this study was detection and analysis of disseminated tumor cells in blood of breast cancer patients by expression profiling using disseminated tumor cell (DTC) detection assay.

Material and Methods: Peripheral EDTA-blood (5ml) of patients with primary breast cancer (n=178) was used for DTC analysis. The assay consists of immunomagnetic tumor cell selection using the cell membrane glycoproteins EpCAM and MUC-1 as targets. The immunobead selected cells were used for mRNA isolation and c-DNA synthesis. The breast carcinoma-associated transcripts EpCAM (EP), MUC-1 (MU) and HER-2 (HE) were analysed by multiplex PCR. Claudin7 (C7) was determined by single-round RT-PCR, while cytokeratin 19 (CK), mammaglobin 1 (MG) prostate-specific ets factor (PSE) and survivin (SU) were determined by nested RT-PCR. PCR products were analysed by capillary electrophoresis with the Agilent Bioanalyzer 2100. Specificity of the RT-PCR was confirmed by examination of blood of healthy donors. Sensitivity for every single transcript was adjusted to 2 tumor cells per 5 ml blood.

Results: Tumor-associated transcripts were detected in 34 of 178 (19,7%) patients with primary breast cancer. Tumor-associated transcripts were heterogenously expressed in positive samples. The marker with the highest incidence of 76% was MUC-1. More than one positive marker was found in 13% of the samples. DTC were identified in 17 from 96 (18%) lymph node negative and 17 from 77 (22%) lymph node positive patients.The expression profile is summarized in Figure 1 [Fig. 1].

Discussion: Using a combination of preanalytical immunomagnetic tumor cell enrichment followed by a multigen RT-PCR we describe a sensitive detection system for breast carcinoma cells. In this study a panel of 8 genes overexpressed at high levels in metastatic breast cancer was selected for the identification of disseminated tumor cells in peripheral blood of patients before primary operation. HER-2, Survivin as a unique member of the inhibitor of apotosis protein family, as well as PSE identified in circulating breast cancer cells may serve as a prognostic indicators of tumor progression and could represent valid targets for new individualized therapeutic interventions.