Characterization of breast cancer interstitial fluids by TmT labeling, ltq-orbitrap velos mass spectrometry, and pathway analysis

Cinzia Raso, Carlo Cosentino, Marco Gaspari, Natalia Malara, Xuemei Han, Daniel McClatchy, Sung Kyu Park, Maria Renne, Nuria Vadalà, Ubaldo Prati, Giovanni Cuda, Vincenzo Mollace, Francesco Amato, John R. Yates

Research output: Contribution to journalArticlepeer-review


Cancer is currently considered as the end point of numerous genomic and epigenomic mutations and as the result of the interaction of transformed cells within the stromal microenvironment. The present work focuses on breast cancer, one of the most common malignancies affecting the female population in industrialized countries. In this study, we perform a proteomic analysis of bioptic samples from human breast cancer, namely, interstitial fluids and primary cells, normal vs disease tissues, using tandem mass tags (TmT) quantitative mass spectrometry combined with the MudPIT technique. To the best of our knowledge, this work, with over 1700 proteins identified, represents the most comprehensive characterization of the breast cancer interstitial fluid proteome to date. Network analysis was used to identify functionally active networks in the breast cancer associated samples. From the list of differentially expressed genes, we have retrieved the associated functional interaction networks. Many different signaling pathways were found activated, strongly linked to invasion, metastasis development, proliferation, and with a significant cross-talking rate. This pilot study presents evidence that the proposed quantitative proteomic approach can be applied to discriminate between normal and tumoral samples and for the discovery of yet unknown carcinogenesis mechanisms and therapeutic strategies.

Original languageEnglish
Pages (from-to)3199-3210
Number of pages12
JournalJournal of Proteome Research
Issue number6
Publication statusPublished - Jun 1 2012


  • breast cancer
  • Cytoscape
  • interstitial fluid
  • LC-MS/MS
  • MudPIT
  • pathway analysis
  • tandem mass tags

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)
  • Medicine(all)


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