Proteomic analysis identifies differentially expressed proteins after HDAC vorinostat and EGFR inhibitor gefitinib treatments in Hep-2 cancer cells

Laura Bianchi, Francesca Bruzzese, Alessandra Leone, Assunta Gagliardi, Michele Puglia, Elena Di Gennaro, Monia Rocco, Anna Gimigliano, Biagio Pucci, Alessandro Armini, Luca Bini, Alfredo Budillon

Research output: Contribution to journalArticle

Abstract

Several solid tumors are characterized by poor prognosis and few effective treatment options, other than palliative chemotherapy in the recurrent/metastatic setting. Epidermal growth factor receptor (EGFR) has been considered an important anticancer target because it is involved in the development and progression of several solid tumors; however, only a subset of patients show a clinically meaningful response to EGFR inhibition, particularly to EGFR tyrosine kinase inhibitors such as gefitinib. We have recently demonstrated synergistic antitumor effect of the histone deacetylase inhibitor vorinostat combined with gefitinib. To further characterize the interaction between these two agents, cellular extracts from Hep-2 cancer cells that were untreated or treated for 24h with either vorinostat or gefitinib alone or with a vorinostat/gefitinib combination were analyzed using 2-D DIGE. Software analysis using DeCyder was performed, and numerous differentially expressed protein spots were visualized between the four examined settings. Using MALDI-TOF MS and ESI-Ion trap MS/MS, several differentially expressed proteins were identified; some of these were validated by Western blotting. Finally, a pathway analysis of experimental data performed using MetaCore highlighted a relevant relationship between the identified proteins and additional potential effectors. In conclusion, we performed a comprehensive analysis of proteins regulated by vorinostat and gefitinib, alone and in combination, providing a useful insight into their mechanisms of action as well as their synergistic interaction.

Original languageEnglish
Pages (from-to)3725-3742
Number of pages18
JournalProteomics
Volume11
Issue number18
DOIs
Publication statusPublished - Sep 2011

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Keywords

  • 2-D DIGE
  • Cell biology
  • Gefitinib
  • Histone deacetylase
  • MS
  • Vorinostat

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

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