2D-DIGE Analysis of Liver Disease in Mice

Abstract

Hepatocellular carcinoma (HCC) is the major type of primary liver cancer. In this chapter, we describe our routine two-dimensional difference gel electrophoresis (2D-DIGE) workflow for analysis of mouse liver tissue in physiological conditions, as well as of mouse HCC. 2D-DIGE still constitutes a valuable comparative proteomics technique, not only providing information on global protein expression in a sample but also on potential posttranslational protein modifications, occurrence of protein degradation fragments, and the existence of protein isoforms. Thus, 2D-DIGE analysis provides highly complementary data to non-gel-based shotgun mass spectrometry (MS) methods (e.g., liquid chromatography (LC)-MS/MS)-allowing, for example, identification of novel protein biomarkers for HCC or increasing insights into the molecular mechanisms underlying hepatocarcinogenesis.

Keywords: Comparative proteomics; Hepatocellular carcinoma (HCC); Liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS); Liver cancer; Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS/MS); Mouse tissue; Two-dimensional difference gel electrophoresis (2D-DIGE).