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
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
19053527
DOI
10.1021/pr800622b
PII: 10.1021/pr800622b
Knihovny.cz E-resources
- MeSH
- Biopsy MeSH
- Early Detection of Cancer MeSH
- Chromatography, Ion Exchange methods MeSH
- Chromatography, Liquid methods MeSH
- Cations MeSH
- Humans MeSH
- Lymphatic Metastasis MeSH
- Neoplasm Metastasis MeSH
- Molecular Weight MeSH
- Biomarkers, Tumor metabolism MeSH
- Breast Neoplasms metabolism MeSH
- Proteome MeSH
- Proteomics methods MeSH
- Salts chemistry MeSH
- Tandem Mass Spectrometry methods MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Cations MeSH
- Biomarkers, Tumor MeSH
- Proteome MeSH
- Salts MeSH
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 < or = 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.
References provided by Crossref.org
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