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

• Authors: BOUCHAL Pavel; ROUMELIOTIS Theodoros; HRSTKA Roman; NENUTIL Rudolf; VOJTĚŠEK Bořivoj; GARBIS Spiros D.
• Year of publication: 2008
• Type: Conference abstract
• MU Faculty or unit: Faculty of Medicine
• Description: The present pilot study constitutes a proof-of-principle in the use of proteomic method incorporating 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) for the comparative analysis of two low-grade breast primary tumor tissues with and without metastases and metastasis in lymph node relative to the nonmetastatic tumor type. 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 low-grade 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 with good intra-assay correlation.

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