The University of Southampton
University of Southampton Institutional Repository

Biomarker discovery in low-grade breast cancer using isobaric stable isotope tags and two-dimensional liquid chromatography-tandem mass spectrometry (iTRAQ-2DLC-MS/MS) based quantitative proteomic analysis

Biomarker discovery in low-grade breast cancer using isobaric stable isotope tags and two-dimensional liquid chromatography-tandem mass spectrometry (iTRAQ-2DLC-MS/MS) based quantitative proteomic analysis
Biomarker discovery in low-grade breast cancer using isobaric stable isotope tags and two-dimensional liquid chromatography-tandem mass spectrometry (iTRAQ-2DLC-MS/MS) based quantitative proteomic analysis
The present pilot study constitutes a proof-of-principle in the use of a quantitative LC-MS/MS based proteomic method for the comparative analysis of representative low-grade breast primary tumor tissues with and without metastases and metastasis in lymph node relative to the nonmetastatic tumor type. The study method incorporated iTRAQ stable isotope labeling, two-dimensional liquid chromatography, nanoelectrospray ionization and high resolution tandem mass spectrometry using the hybrid QqTOF platform (iTRAQ-2DLC-MS/MS). The principal aims of this study were (1) to define the protein spectrum obtainable using this approach, and (2) to highlight potential candidates for verification and validation studies focused on biomarkers involved in metastatic processes in breast cancer. The study resulted in the reproducible identification of 605 nonredundant proteins (p ? 0.05). A quantitative comparison revealed 3/3 proteins with significantly increased/decreased level in metastatic primary tumor and 13/6 proteins with increased/decreased level in lymph node metastasis compared to nonmetastatic primary tumor (p < 0.01). Changes in selected differentially expressed proteins were verified with qRT-PCR. Although our pilot scale study does not warrant general biological conclusions, the synergic regulation of some proteins with related function (e.g., heme binding proteins, proteins of energetic metabolism, interferon induced proteins, proteins with adhesive function) determined in our sample set reflects the ability of our method in providing biologically meaningful data. The main conclusion from this pilot study was that our quantitative proteomic method constitutes a novel way of analyzing cancerous breast tissue biopsy samples that can be extended as part of a larger scale biomarker discovery program.

1535-3893
362-373
Bouchal, Pavel
49baebee-1916-457d-983d-7f93a348c601
Roumeliotis, Theodoros
f1284c98-b5eb-483e-9416-594c678e62fd
Hrstka, Roman
b08b0f63-7d4c-4c7d-a90f-454db4dd57ed
Nenutil, Rudolf
21fcec7b-87ec-44a8-bf1c-1e673cb1bfed
Vojtesek, Borivoj
3a9074e5-0f44-4604-bfba-ca0426cb926b
Garbis, Spiros D.
7067fd19-50c9-4d42-9611-f370289470bd
Bouchal, Pavel
49baebee-1916-457d-983d-7f93a348c601
Roumeliotis, Theodoros
f1284c98-b5eb-483e-9416-594c678e62fd
Hrstka, Roman
b08b0f63-7d4c-4c7d-a90f-454db4dd57ed
Nenutil, Rudolf
21fcec7b-87ec-44a8-bf1c-1e673cb1bfed
Vojtesek, Borivoj
3a9074e5-0f44-4604-bfba-ca0426cb926b
Garbis, Spiros D.
7067fd19-50c9-4d42-9611-f370289470bd

Bouchal, Pavel, Roumeliotis, Theodoros, Hrstka, Roman, Nenutil, Rudolf, Vojtesek, Borivoj and Garbis, Spiros D. (2009) Biomarker discovery in low-grade breast cancer using isobaric stable isotope tags and two-dimensional liquid chromatography-tandem mass spectrometry (iTRAQ-2DLC-MS/MS) based quantitative proteomic analysis. Journal of Proteome Research, 8 (1), 362-373. (doi:10.1021/pr800622b).

Record type: Article

Abstract

The present pilot study constitutes a proof-of-principle in the use of a quantitative LC-MS/MS based proteomic method for the comparative analysis of representative low-grade breast primary tumor tissues with and without metastases and metastasis in lymph node relative to the nonmetastatic tumor type. The study method incorporated iTRAQ stable isotope labeling, two-dimensional liquid chromatography, nanoelectrospray ionization and high resolution tandem mass spectrometry using the hybrid QqTOF platform (iTRAQ-2DLC-MS/MS). The principal aims of this study were (1) to define the protein spectrum obtainable using this approach, and (2) to highlight potential candidates for verification and validation studies focused on biomarkers involved in metastatic processes in breast cancer. The study resulted in the reproducible identification of 605 nonredundant proteins (p ? 0.05). A quantitative comparison revealed 3/3 proteins with significantly increased/decreased level in metastatic primary tumor and 13/6 proteins with increased/decreased level in lymph node metastasis compared to nonmetastatic primary tumor (p < 0.01). Changes in selected differentially expressed proteins were verified with qRT-PCR. Although our pilot scale study does not warrant general biological conclusions, the synergic regulation of some proteins with related function (e.g., heme binding proteins, proteins of energetic metabolism, interferon induced proteins, proteins with adhesive function) determined in our sample set reflects the ability of our method in providing biologically meaningful data. The main conclusion from this pilot study was that our quantitative proteomic method constitutes a novel way of analyzing cancerous breast tissue biopsy samples that can be extended as part of a larger scale biomarker discovery program.

Full text not available from this repository.

More information

e-pub ahead of print date: 20 November 2008
Published date: January 2009
Organisations: Cancer Sciences

Identifiers

Local EPrints ID: 341680
URI: https://eprints.soton.ac.uk/id/eprint/341680
ISSN: 1535-3893
PURE UUID: 57576bb4-f046-4e1b-a926-509ba8cd4849

Catalogue record

Date deposited: 02 Aug 2012 15:14
Last modified: 18 Jul 2017 05:33

Export record

Altmetrics

Contributors

Author: Pavel Bouchal
Author: Theodoros Roumeliotis
Author: Roman Hrstka
Author: Rudolf Nenutil
Author: Borivoj Vojtesek

University divisions

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of https://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×