Comparative proteomic analysis of paclitaxel sensitive A2780 epithelial ovarian cancer cell line and its resistant counterpart A2780TC1 by 2D-DIGE: The role of ERp57

Lucia Cicchihitti, Michela Di Michele, Andrea Urbani, Cristiano Ferlini, Maria Benedetta Donati, Giovanni Scambia, Domenico Rotilio

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

Epithelial ovarian cancer is the leading cause of gynecological cancer mortality. Despite good response to surgery and initial chemotherapy, chemoresistance occurrence represents a major obstacle to a successful therapy. To better understand biological mechanisms at the basis of paclitaxel resistance, a comparative proteomic approach based on DIGE coupled with mass spectrometry (MALDI-TOF and LC-MS/MS) was applied to the human epithelial ovarian cancer cell lines A2780 and its paclitaxel resistant counterpart A2780TC1. Most of the differentially expressed proteins between the two cell lines belong to the class of stress response (29%), metabolism (21%), and cell cycle and apoptosis (17%). We focused on proteins which were most strongly modulated by paclitaxel resistance and in particular on the disulphide isomerase ERp57, which may represent a chemoresistance biomarker. ERp57 was found to interact with class III β-tubulin (TUBB3), involved in paclitaxel resistance in ovarian and other cancers. Moreover, we demonstrated a novel localization of this protein in cytoskeleton and described that ERp57/TUBB3 interaction occurs also in the nuclear compartment and in association with a multimeric complex formed by nucleolin, nucleophosmin, hnRNPK, and mortalin. Our data suggest that ERp57 plays an important role in chemoresistance mechanisms in ovarian cancer by modulating the attachment of microtubules to chromosomes following paclitaxel treatment through its interaction with TUBB3.

Original languageEnglish
Pages (from-to)1902-1912
Number of pages11
JournalJournal of Proteome Research
Volume8
Issue number4
DOIs
Publication statusPublished - Apr 3 2009

Fingerprint

Two-Dimensional Difference Gel Electrophoresis
Paclitaxel
Proteomics
Cells
Cell Line
Ovarian Neoplasms
Protein Disulfide-Isomerases
Proteins
Chemotherapy
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Biomarkers
Tubulin
Chromosomes
Cytoskeleton
Metabolism
Microtubules
Surgery
Mass spectrometry
Mass Spectrometry
Cell Cycle

Keywords

  • 2D-DIGE
  • Biomarker
  • Chemoresistance
  • ERp57
  • Mass spectrometry
  • Ovarian cancer
  • Paclitaxel
  • Proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Comparative proteomic analysis of paclitaxel sensitive A2780 epithelial ovarian cancer cell line and its resistant counterpart A2780TC1 by 2D-DIGE : The role of ERp57. / Cicchihitti, Lucia; Michele, Michela Di; Urbani, Andrea; Ferlini, Cristiano; Donati, Maria Benedetta; Scambia, Giovanni; Rotilio, Domenico.

In: Journal of Proteome Research, Vol. 8, No. 4, 03.04.2009, p. 1902-1912.

Research output: Contribution to journalArticle

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abstract = "Epithelial ovarian cancer is the leading cause of gynecological cancer mortality. Despite good response to surgery and initial chemotherapy, chemoresistance occurrence represents a major obstacle to a successful therapy. To better understand biological mechanisms at the basis of paclitaxel resistance, a comparative proteomic approach based on DIGE coupled with mass spectrometry (MALDI-TOF and LC-MS/MS) was applied to the human epithelial ovarian cancer cell lines A2780 and its paclitaxel resistant counterpart A2780TC1. Most of the differentially expressed proteins between the two cell lines belong to the class of stress response (29{\%}), metabolism (21{\%}), and cell cycle and apoptosis (17{\%}). We focused on proteins which were most strongly modulated by paclitaxel resistance and in particular on the disulphide isomerase ERp57, which may represent a chemoresistance biomarker. ERp57 was found to interact with class III β-tubulin (TUBB3), involved in paclitaxel resistance in ovarian and other cancers. Moreover, we demonstrated a novel localization of this protein in cytoskeleton and described that ERp57/TUBB3 interaction occurs also in the nuclear compartment and in association with a multimeric complex formed by nucleolin, nucleophosmin, hnRNPK, and mortalin. Our data suggest that ERp57 plays an important role in chemoresistance mechanisms in ovarian cancer by modulating the attachment of microtubules to chromosomes following paclitaxel treatment through its interaction with TUBB3.",
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