Volume 10 Supplement 2

Breast Cancer Research 2008

Open Access

Detection of gene amplification in matched tumour and plasma DNA from breast cancer patients by quantitative PCR

  • K Page1,
  • N Hava2,
  • MJ Slade2,
  • RA Walker1,
  • RC Coombes2 and
  • JA Shaw1
Breast Cancer Research200810(Suppl 2):P70

https://doi.org/10.1186/bcr1954

Published: 13 May 2008

Background

The aim of the present study was to determine whether tumour-specific gene amplification is detectable in circulating cell-free DNA isolated from the plasma of breast cancer patients. Four loci, known to show amplification in breast cancers, were chosen for investigation of paired plasma and tumour DNA: Her-2 and C35 on chromosome 17, and FGFR1 and RAB11f1P1, which map to chromosome 8. Both gene pairs are frequently, but not exclusively, coamplified in breast tumours.

Methods

Lymphocyte, tumour and plasma DNA from 21 unselected breast cancer cases were analysed for amplification by a real-time quantitative PCR assay. Primers and a minor groove binder (MGB) TaqMan probe were targeted to the four genes and to an unamplified reference gene selected from each of the two intervals (CNTNAP1 and UNC5ND, respectively). A normal lymphocyte DNA sample was included in each experiment to examine interassay reproducibility and as an experimental calibrator. A ratio above 2.0 was regarded as positive for gene amplification [1].

Results

Amplification was detected in tumour and plasma DNA at all four loci, with frequencies ranging from 19.1% to 52.3%. Five of 21 cases showed concordant amplification of Her-2 in plasma and tumour DNA, compared with four, one and two of 21 cases for C35, FGFR1 and RAB11f1P1 amplification, respectively. No amplification was seen in the lymphocyte DNA samples.

Conclusion

Taken together, these data confirm that tumour-specific amplification, including Her-2 amplification, is detectable in circulating cell-free DNA isolated from the plasma of breast cancer patients. This suggests the possibility of developing a rapid and simple blood screening tool for identification of gene amplification in breast cancer cases.

Authors’ Affiliations

(1)
Department of Cancer Studies and Molecular Medicine, University of Leicester
(2)
Department of Cancer Medicine, Imperial College, Hammersmith Hospital

References

  1. Kulka J, Tokes AM, Kaposi-Novak P, Udvarhelyi N, Keller A, Schaff Z: Detection of HER-2/neu gene amplification in breast carcinomas using quantitative real-time PCR – a comparison with immunohistochemical and FISH results. Pathol Oncol Res. 2006, 12: 197-204.View ArticlePubMedGoogle Scholar

Copyright

© BioMed Central Ltd 2008

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