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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Editorial Board Member: Gillery, Philippe / Kazmierczak, Steven / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Schlattmann, Peter / Whitfield, John B.

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High-performance metabolic marker assessment in breast cancer tissue by mass spectrometry

Maria Chiara Mimmi1, 2, a / Paola Picotti1, 2, a, b / Alessandra Corazza1, 2 / Elena Betto1, 2 / Carlo E. Pucillo1, 2 / Laura Cesaratto1 / Carla Cedolini3 / Viviana Londero3 / Chiara Zuiani3, 4 / Massimo Bazzocchi3, 4 / 1, 2

1Dipartimento di Scienze e Tecnologie Biomediche, Università di Udine, Udine, Italy

2MATI Centre of Excellence, Università di Udine, Udine, Italy

3Azienda Ospedaliera Universitaria di Udine (AOUD), Udine, Italy

4Dipartimento di Ricerche Mediche e Morfologiche, Università di Udine, Udine, Italy

aM.C. Mimmi and P. Picotti contributed equally.

bP.P. Present address: Institute of Molecular Systems Biology, ETH Zurich, Zurich 8093, Switzerland.

Corresponding author: Gennaro Esposito, Dipartimento Scienze e Tecnologie Biomediche, Università di Udine, P.le Kolbe, 4, 33100 Udine, Italy Phone: +39 0432494321, Fax: +39 0432494301

Citation Information: Clinical Chemistry and Laboratory Medicine. Volume 49, Issue 2, Pages 317–324, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: 10.1515/CCLM.2011.060, December 2010

Publication History

Received:
2010-02-12
Accepted:
2010-08-17
Published Online:
2010-12-14

Abstract

Background: The identification of reliable markers for diagnosis of breast cancer has been thoroughly addressed by metabolic profiling using nuclear magnetic resonance (NMR) spectroscopy or imaging. Several clear diagnostic indicators have emerged using either in vitro analysis of tissue extracts, ex vivo analysis of biopsies or in vivo direct spectral observations. Most of the breast cancer characteristic metabolites could be assayed by mass spectrometry (MS) to exploit the superior sensitivity of this technique and therefore reduce the traumatic impact of current biopsy procedures.

Methods: Following extraction, aqueous metabolite mixtures were obtained that were submitted to liquid-chromatography, electrospray-ionization, mass spectrometry (LC/ESI-MS) analysis to estimate the content of choline (Cho) and its phosphorylated derivatives, phosphocholine (PCho) and glycerophosphocholine (GPCho). The determinations were performed using 10 samples from breast tissue biopsies, surgical specimens and one single sample of a hepatic metastasis. In addition, some measurements were also repeated using high-resolution 1H NMR spectroscopy to complement the mass spectrometry results.

Results: The contents of Cho, PCho and GPCho in breast tissue extracts were estimated by LC/ESI-MS based on standard compound calibration curves. Sharply increased ratios of phosphorylated-to-unphosphorylated metabolites, PCho/ Cho and (PCho+GPCho)/Cho, were observed in all tumor samples, although without discrimination between benign and malignant lesions, contrary to samples from healthy individuals and from those with fibrocystic disease.

Conclusions: The assessment of breast cancer markers by LC/ESI-MS is feasible and diagnostically valuable. In addition to high sensitivity, the approach also shows a resolution advantage for assaying choline derivatives compared to NMR, and could complement the latter.

Keywords: breast cancer; breast tissue metabonomics; choline metabolites; mass spectrometry metabonomics; tumor marker detection by nuclear magnetic resonance and mass spectrometry

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