title: 2D-PAGE of ovarian cancer: analysis of soluble and insoluble fractions using medium-range immobilized pH gradients.
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.
authors.affiliation: Department of Obstetrics and Gynaecology, University of Melbourne, Victoria, Australia. ratanal@unimelb.edu.au
authors.forename: Ratana
authors.initials: R
authors.lastname: Lim
authors.forename: Martha
authors.initials: M
authors.lastname: Lappas
authors.forename: Nuzhat
authors.initials: N
authors.lastname: Ahmed
authors.forename: Michael
authors.initials: M
authors.lastname: Permezel
authors.forename: Michael A
authors.initials: MA
authors.lastname: Quinn
authors.forename: Gregory E
authors.initials: GE
authors.lastname: Rice
mesh_terms: D005591:Chemical Fractionation; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D000091342:Endoplasmic Reticulum Chaperone BiP; D005260:Female; D006360:Heat-Shock Proteins / Q000032:analysis / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009363:Neoplasm Proteins / Q000032:analysis* / Q000378:metabolism; D009367:Neoplasm Staging; D010051:Ovarian Neoplasms / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology*; D000091402:Prohibitins; D040901:Proteomics; D012097:Repressor Proteins / Q000032:analysis / Q000378:metabolism; D012995:Solubility
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; D009363:Neoplasm Proteins; D000091402:Prohibitins; D012097:Repressor Proteins