Cargando…
Absolute Quantification of Choline-Related Biomarkers in Breast Cancer Biopsies by Liquid Chromatography Electrospray Ionization Mass Spectrometry
It has been repeatedly demonstrated that choline metabolism is altered in a wide variety of cancers. In breast tumours, the choline metabolite profile is characterized by an elevation of phosphocholine and total choline-compounds. This pattern is increasingly being exploited as biomarker in cancer d...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
IOS Press
2013
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4605703/ https://www.ncbi.nlm.nih.gov/pubmed/24036926 http://dx.doi.org/10.3233/ACP-130082 |
_version_ | 1782395244617138176 |
---|---|
author | Mimmi, Maria Chiara Finato, Nicoletta Pizzolato, Gloria Beltrami, Carlo Alberto Fogolari, Federico Corazza, Alessandra Esposito, Gennaro |
author_facet | Mimmi, Maria Chiara Finato, Nicoletta Pizzolato, Gloria Beltrami, Carlo Alberto Fogolari, Federico Corazza, Alessandra Esposito, Gennaro |
author_sort | Mimmi, Maria Chiara |
collection | PubMed |
description | It has been repeatedly demonstrated that choline metabolism is altered in a wide variety of cancers. In breast tumours, the choline metabolite profile is characterized by an elevation of phosphocholine and total choline-compounds. This pattern is increasingly being exploited as biomarker in cancer diagnosis. The majority of in vitro metabolomics studies, for biomarkers quantification in cell cultures or tissues, entail proton NMR spectroscopy. Although many “targeted” approaches have been proposed to quantify metabolites from standard one-dimensional (1D) NMR experiments, the task is often made difficult by the high degree of overlap characterizing (1)H NMR spectra of biological samples. Here we present an optimized protocol for tissue extraction and absolute quantification of choline, phosphocholine and glycerophosphocholine by means of liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS). The selected chromatographic separation system with a HILIC (hydrophilic interaction chromatography) amide column effectively separates free choline and its phopshorylated derivatives, contrary to failure observed using standard reversed-phase chromatography. The metabolite absolute quantification is based on external calibration with commercial standards, and is validated by a parallel 1D proton NMR analysis. The LC-MS/NMR analysis is applied to three breast carcinoma specimens obtained by surgical excision, each one accompanied by a control tissue sample taken outside the tumor margin. The metabolite concentrations measured are in good agreement with previous results on metabolic profile changes of breast cancer. Each of the three cancerous biopsies, when compared with the control tissue, exhibit a highly increased levels phosphocholine, total choline and phosphocholine/glycerophosphocholine ratio. |
format | Online Article Text |
id | pubmed-4605703 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | IOS Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-46057032015-12-13 Absolute Quantification of Choline-Related Biomarkers in Breast Cancer Biopsies by Liquid Chromatography Electrospray Ionization Mass Spectrometry Mimmi, Maria Chiara Finato, Nicoletta Pizzolato, Gloria Beltrami, Carlo Alberto Fogolari, Federico Corazza, Alessandra Esposito, Gennaro Anal Cell Pathol (Amst) Other It has been repeatedly demonstrated that choline metabolism is altered in a wide variety of cancers. In breast tumours, the choline metabolite profile is characterized by an elevation of phosphocholine and total choline-compounds. This pattern is increasingly being exploited as biomarker in cancer diagnosis. The majority of in vitro metabolomics studies, for biomarkers quantification in cell cultures or tissues, entail proton NMR spectroscopy. Although many “targeted” approaches have been proposed to quantify metabolites from standard one-dimensional (1D) NMR experiments, the task is often made difficult by the high degree of overlap characterizing (1)H NMR spectra of biological samples. Here we present an optimized protocol for tissue extraction and absolute quantification of choline, phosphocholine and glycerophosphocholine by means of liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS). The selected chromatographic separation system with a HILIC (hydrophilic interaction chromatography) amide column effectively separates free choline and its phopshorylated derivatives, contrary to failure observed using standard reversed-phase chromatography. The metabolite absolute quantification is based on external calibration with commercial standards, and is validated by a parallel 1D proton NMR analysis. The LC-MS/NMR analysis is applied to three breast carcinoma specimens obtained by surgical excision, each one accompanied by a control tissue sample taken outside the tumor margin. The metabolite concentrations measured are in good agreement with previous results on metabolic profile changes of breast cancer. Each of the three cancerous biopsies, when compared with the control tissue, exhibit a highly increased levels phosphocholine, total choline and phosphocholine/glycerophosphocholine ratio. IOS Press 2013 2013-09-13 /pmc/articles/PMC4605703/ /pubmed/24036926 http://dx.doi.org/10.3233/ACP-130082 Text en Copyright © 2013 Hindawi Publishing Corporation and the authors. |
spellingShingle | Other Mimmi, Maria Chiara Finato, Nicoletta Pizzolato, Gloria Beltrami, Carlo Alberto Fogolari, Federico Corazza, Alessandra Esposito, Gennaro Absolute Quantification of Choline-Related Biomarkers in Breast Cancer Biopsies by Liquid Chromatography Electrospray Ionization Mass Spectrometry |
title | Absolute Quantification of Choline-Related Biomarkers in Breast Cancer Biopsies by Liquid Chromatography Electrospray Ionization Mass Spectrometry |
title_full | Absolute Quantification of Choline-Related Biomarkers in Breast Cancer Biopsies by Liquid Chromatography Electrospray Ionization Mass Spectrometry |
title_fullStr | Absolute Quantification of Choline-Related Biomarkers in Breast Cancer Biopsies by Liquid Chromatography Electrospray Ionization Mass Spectrometry |
title_full_unstemmed | Absolute Quantification of Choline-Related Biomarkers in Breast Cancer Biopsies by Liquid Chromatography Electrospray Ionization Mass Spectrometry |
title_short | Absolute Quantification of Choline-Related Biomarkers in Breast Cancer Biopsies by Liquid Chromatography Electrospray Ionization Mass Spectrometry |
title_sort | absolute quantification of choline-related biomarkers in breast cancer biopsies by liquid chromatography electrospray ionization mass spectrometry |
topic | Other |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4605703/ https://www.ncbi.nlm.nih.gov/pubmed/24036926 http://dx.doi.org/10.3233/ACP-130082 |
work_keys_str_mv | AT mimmimariachiara absolutequantificationofcholinerelatedbiomarkersinbreastcancerbiopsiesbyliquidchromatographyelectrosprayionizationmassspectrometry AT finatonicoletta absolutequantificationofcholinerelatedbiomarkersinbreastcancerbiopsiesbyliquidchromatographyelectrosprayionizationmassspectrometry AT pizzolatogloria absolutequantificationofcholinerelatedbiomarkersinbreastcancerbiopsiesbyliquidchromatographyelectrosprayionizationmassspectrometry AT beltramicarloalberto absolutequantificationofcholinerelatedbiomarkersinbreastcancerbiopsiesbyliquidchromatographyelectrosprayionizationmassspectrometry AT fogolarifederico absolutequantificationofcholinerelatedbiomarkersinbreastcancerbiopsiesbyliquidchromatographyelectrosprayionizationmassspectrometry AT corazzaalessandra absolutequantificationofcholinerelatedbiomarkersinbreastcancerbiopsiesbyliquidchromatographyelectrosprayionizationmassspectrometry AT espositogennaro absolutequantificationofcholinerelatedbiomarkersinbreastcancerbiopsiesbyliquidchromatographyelectrosprayionizationmassspectrometry |