Dithiothreitol-based protein equalization technology to unravel biomarkers for bladder cancer

José E. Araújo, H. López-Fernández, M. S. Diniz, Pedro M. Baltazar, Luís Campos Pinheiro, Fernando Calais da Silva, Mylène Carrascal, Paula Videira, H. M. Santos, J. L. Capelo

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Abstract

This study aimed to assess the benefits of dithiothreitol (DTT)-based sample treatment for protein equalization to assess potential biomarkers for bladder cancer. The proteome of plasma samples of patients with bladder carcinoma, patients with lower urinary tract symptoms (LUTS) and healthy volunteers, was equalized with dithiothreitol (DTT) and compared. The equalized proteomes were interrogated using two-dimensional gel electrophoresis and matrix assisted laser desorption ionization time of flight mass spectrometry. Six proteins, namely serum albumin, gelsolin, fibrinogen gamma chain, Ig alpha-1 chain C region, Ig alpha-2 chain C region and haptoglobin, were found dysregulated in at least 70% of bladder cancer patients when compared with a pool of healthy individuals. One protein, serum albumin, was found overexpressed in 70% of the patients when the equalized proteome of the healthy pool was compared with the equalized proteome of the LUTS patients. The pathways modified by the proteins differentially expressed were analyzed using Cytoscape. The method here presented is fast, cheap, of easy application and it matches the analytical minimalism rules as outlined by Halls. Orthogonal validation was done using western-blot. Overall, DTT-based protein equalization is a promising methodology in bladder cancer research.

Original languageEnglish
Pages (from-to)36-46
Number of pages11
JournalTalanta
Volume180
DOIs
Publication statusPublished - 1 Apr 2018

Keywords

  • 2D-SDS-PAGE
  • Bladder cancer
  • LUTS
  • MALDI-TOF-MS
  • Protein equalization
  • S2P

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