2D-PAGE of ovarian cancer: analysis of soluble and insoluble fractions using medium-range immobilized pH gradients

Biochem Biophys Res Commun. 2011 Mar 18;406(3):408-13. doi: 10.1016/j.bbrc.2011.02.056. Epub 2011 Feb 15.

Abstract

Ovarian cancer remains a leading cause of cancer death. A comparative proteomic study was performed on normal ovarian tissue (n=5) and grade 3 ovarian tumours (n=5) to search for differentially expressed proteins. In contrast to other studies, here we extracted proteins in soluble and insoluble protein fractions using commercial kits and also utilised three medium-range IPG strips that encompassed the broad pH range of 3-10 (pH 3-6, 5-8 and 7-10). Protein fractions were compared by 2D-PAGE and MALDI-TOF/TOF-MS. Nineteen differentially expressed proteins were identified: HSP60, Grp78, CK19, EF-Tu, MRLC2, prohibitin, Stress-70 protein, TPI and tubulin α6 were up-regulated in grade 3 tumours whereas annexin A2 and A5, antithrombin-III precursor, CBR1, GSTM2, GSTM3, RALDH1, serum albumin precursor, transthyretin precursor and vimentin were found to be down-regulated in grade 3 ovarian tumours. These proteins are associated with cytoskeleton rearrangement, cell metabolism, tumour suppression function, apoptosis and induction of host response.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chemical Fractionation
  • Cytoskeleton / genetics
  • Cytoskeleton / metabolism
  • Electrophoresis, Gel, Two-Dimensional
  • Endoplasmic Reticulum Chaperone BiP
  • Female
  • Heat-Shock Proteins / analysis
  • Heat-Shock Proteins / metabolism
  • Humans
  • Hydrogen-Ion Concentration
  • Neoplasm Proteins / analysis*
  • Neoplasm Proteins / metabolism
  • Neoplasm Staging
  • Ovarian Neoplasms / chemistry*
  • Ovarian Neoplasms / metabolism
  • Ovarian Neoplasms / pathology*
  • Prohibitins
  • Proteomics*
  • Repressor Proteins / analysis
  • Repressor Proteins / metabolism
  • Solubility

Substances

  • Endoplasmic Reticulum Chaperone BiP
  • HSPA5 protein, human
  • Heat-Shock Proteins
  • Neoplasm Proteins
  • Prohibitins
  • Repressor Proteins