Breast Cancer Research and Treatment

, Volume 146, Issue 1, pp 163–174 | Cite as

Plasma DNA integrity as a biomarker for primary and metastatic breast cancer and potential marker for early diagnosis

  • Dharanija Madhavan
  • Markus Wallwiener
  • Karin Bents
  • Manuela Zucknick
  • Juliane Nees
  • Sarah Schott
  • Katarina Cuk
  • Sabine Riethdorf
  • Andreas Trumpp
  • Klaus Pantel
  • Christof Sohn
  • Andreas Schneeweiss
  • Harald SurowyEmail author
  • Barbara Burwinkel


Circulating or cell-free DNA (cfDNA) has been evaluated as a biomarker in many cancers including breast cancer. In particular, integrity of cfDNA has been shown to be altered in cancers. We have estimated the biomarker potential of cfDNA in primary (PBC) and metastatic breast cancer (MBC). cfDNA integrity (cfDI) and concentration were determined in plasma of 383 individuals, including 82 PBC and 201 MBC cases, as well as 100 healthy controls, by measuring ALU and LINE1 repetitive DNA elements using quantitative PCR. The MBC patient group was further sub-divided into patients with detectable circulating tumour cells (CTCpos-MBC, n = 100) and those without (CTCneg-MBC, n = 101). A hierarchical decrease in cfDI and increase in cfDNA concentration from healthy controls to PBC and further onto MBC patients were observed. Investigation of cfDNA in media of cell lines was in concordance with these results. Combination of cfDI and cfDNA concentration could differentiate PBC cases from controls (area under the curve, AUC = 0.75), MBC cases from controls (AUC = 0.81 for CTCneg-MBC, AUC = 0.93 for CTCpos-MBC), and CTCneg-MBC from CTCpos-MBC cases (AUC = 0.83). cfDI additionally demonstrated a positive correlation to progression-free (HR of 0.46 for ALU, P = 0.0025) and overall survival (HR of 0.15 for ALU and 0.20 for LINE1, P < 0.0001) in MBC, and had lower prediction error than CTC status. Our findings show that reduced cfDI and increased cfDNA concentration can serve as diagnostic markers for PBC and MBC, and cfDI as a prognostic marker for MBC, thereby making them attractive candidates for blood-based multi-marker assays.


Breast cancer Circulating DNA DNA integrity Diagnostic marker Prognostic marker 



Area under the curve


Circulating or cell-free DNA


Cell-free DNA integrity


Circulating tumour cells


Circulating tumour cells positive metastatic breast cancer


Circulating tumour cells negative metastatic breast cancer


Hazard ratio


Metastatic breast cancer


Primary breast cancer


Progression-free survival


Overall survival



We thank the study participants and all our colleagues who helped us with patient recruitment, blood collection and processing. This study was funded and supported by the Dietmar-Hopp Foundation, the University Hospital of Heidelberg, the Helmholtz Society, and the German Cancer Research Center (DKFZ), Heidelberg, Germany.

Supplementary material

10549_2014_2946_MOESM1_ESM.pdf (343 kb)
Supplementary Material (PDF 342 kb)


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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Dharanija Madhavan
    • 1
    • 2
  • Markus Wallwiener
    • 3
    • 4
  • Karin Bents
    • 2
  • Manuela Zucknick
    • 5
  • Juliane Nees
    • 3
    • 4
  • Sarah Schott
    • 3
  • Katarina Cuk
    • 1
    • 2
  • Sabine Riethdorf
    • 6
  • Andreas Trumpp
    • 7
    • 8
  • Klaus Pantel
    • 6
  • Christof Sohn
    • 3
  • Andreas Schneeweiss
    • 3
    • 4
  • Harald Surowy
    • 1
    • 2
    Email author
  • Barbara Burwinkel
    • 1
    • 2
  1. 1.Molecular Epidemiology, C080German Cancer Research Center (DKFZ)HeidelbergGermany
  2. 2.Molecular Biology of Breast Cancer, Department of Gynecology and ObstetricsUniversity of HeidelbergHeidelbergGermany
  3. 3.Department of Gynecology and ObstetricsUniversity of HeidelbergHeidelbergGermany
  4. 4.National Center for Tumor DiseasesUniversity of HeidelbergHeidelbergGermany
  5. 5.Division of BiostatisticsGerman Cancer Research Center (DKFZ)HeidelbergGermany
  6. 6.Department of Tumor BiologyUniversity Hospital Hamburg-EppendorfHamburgGermany
  7. 7.Hi-STEM-Heidelberg Institute for Stem Cell Technology and Experimental Medicine, GmbHHeidelbergGermany
  8. 8.Division of Stem Cells and CancerGerman Cancer Research Center (DKFZ)HeidelbergGermany

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