DIGE-based protein expression analysis of B[a]P-exposed hepatoma cells reveals a complex stress response including alterations in oxidative stress, cell cycle control, and cytoskeleton motility at toxic and subacute concentrations

J Proteome Res. 2011 Feb 4;10(2):379-93. doi: 10.1021/pr100723d. Epub 2010 Dec 21.


Although the effects of high concentrations of the carcinogen benzo[a]pyrene (B[a]P) have been studied extensively, little is known about its effects at subacute toxic concentrations, which are typical for environmental pollutants. We exposed murine Hepa1c1c7 cells to a toxic concentration (5 μM) and a subacute concentration (50 nM) of B[a]P over a period of 2-24 h to differentiate between acute and pseudochronic effects and conducted a time-course analysis of B[a]P-influenced protein expression by DIGE. In total, a set of 120 spots were found to be significantly altered due to B[a]P exposure of which 112 were subsequently identified by mass spectrometry. Clustering and principal component analysis were conducted to identify sets of proteins responding in a concerted manner to the exposure. Our results indicate an immediate response to the contaminant at the protein level and demonstrate that B[a]P exposure alters the cellular response by disturbing proteins involved in oxidative stress, cell cycle regulation, apoptosis, and cytoskeleton organization. Furthermore, network analysis of protein-protein interactions revealed a complex network of interacting, B[a]P-regulated proteins mostly belonging to the cytoskeleton organization and several signal transduction pathways.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Benzo(a)pyrene / toxicity*
  • Carcinoma, Hepatocellular / metabolism*
  • Cell Cycle / drug effects*
  • Cell Line, Tumor
  • Cytoskeleton / metabolism
  • Databases, Protein
  • Dose-Response Relationship, Drug
  • Electrophoresis, Gel, Two-Dimensional
  • Flow Cytometry
  • Humans
  • Liver Neoplasms / metabolism
  • Mice
  • Oxidative Stress / drug effects*
  • Protein Interaction Mapping
  • Proteome / metabolism*
  • Signal Transduction / drug effects
  • Toxicity Tests


  • Proteome
  • Benzo(a)pyrene