Combined gene expression and proteomic analysis of EGF induced apoptosis in A431 cells suggests multiple pathways trigger apoptosis

Apoptosis. 2013 Nov;18(11):1291-1305. doi: 10.1007/s10495-013-0887-6.

Abstract

A431 cells, derived from epidermoid carcinoma, overexpress the epidermal growth factor receptor (EGFR) and when treated with a high dose of EGF will undergo apoptosis. We exploited microarray and proteomics techniques and network prediction to study the regulatory mechanisms of EGF-induced apoptosis in A431 cells. We observed significant changes in gene expression in 162 genes, approximately evenly split between pro-apoptotic and anti-apoptotic genes and identified 30 proteins from the proteomic data that had either pro or anti-apoptotic annotation. Our correlation analysis of gene expression and proteome modeled a number of distinct sub-networks that are associated with the onset of apoptosis, allowing us to identify specific pathways and components. These include components of the interferon signalling pathway, and down stream components, including cytokines and suppressors of cytokine signalling. A central component of almost all gene expression sub-networks identified was TP53, which is mutated in A431 cells, and was down regulated. This down regulation of TP53 appeared to be correlated with proteomic sub-networks of cytoskeletal or cell adhesion components that might induce apoptosis by triggering cytochrome C release. Of the only three genes also differentially expressed as proteins, only serpinb1 had a known association with apoptosis. We confirmed that up regulation and cleavage of serpinb1 into L-DNAaseII was correlated with the induction of apoptosis. It is unlikely that a single pathway, but more likely a combination of pathways is needed to trigger EGF induced apoptosis in A431cells.

MeSH terms

  • Apoptosis / drug effects*
  • Apoptosis / genetics
  • Cell Adhesion / drug effects
  • Cell Line, Tumor
  • Cytochromes c / metabolism
  • Cytoskeleton / drug effects
  • Cytoskeleton / genetics
  • Cytoskeleton / metabolism
  • Cytoskeleton / ultrastructure
  • Epidermal Growth Factor / pharmacology*
  • Epithelial Cells
  • ErbB Receptors / genetics*
  • ErbB Receptors / metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic / drug effects*
  • Humans
  • Interferons / genetics
  • Interferons / metabolism
  • Oligonucleotide Array Sequence Analysis
  • Proteomics
  • Serpins / genetics
  • Serpins / metabolism
  • Signal Transduction / drug effects*
  • Tumor Suppressor Protein p53 / genetics*
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • Serpins
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • SERPINB1 protein, human
  • Epidermal Growth Factor
  • Cytochromes c
  • Interferons
  • EGFR protein, human
  • ErbB Receptors