title: Proteomic Analysis of Matched Formalin-Fixed, Paraffin-Embedded Specimens in Patients with Advanced Serous Ovarian Carcinoma.
abstract: OBJECTIVE
  The biology of high grade serous ovarian carcinoma (HGSOC) is poorly understood. Little has been reported on intratumoral homogeneity or heterogeneity of primary HGSOC tumors and their metastases. We evaluated the global protein expression profiles of paired primary and metastatic HGSOC from formalin-fixed, paraffin-embedded (FFPE) tissue samples.
  
  
  METHODS
  After IRB approval, six patients with advanced HGSOC were identified with tumor in both ovaries at initial surgery. Laser capture microdissection (LCM) was used to extract tumor for protein digestion. Peptides were extracted and analyzed by reversed-phase liquid chromatography coupled to a linear ion trap mass spectrometer. Tandem mass spectra were searched against the UniProt human protein database. Differences in protein abundance between samples were assessed and analyzed by Ingenuity Pathway Analysis software. Immunohistochemistry (IHC) for select proteins from the original and an additional validation set of five patients was performed.
  
  
  RESULTS
  Unsupervised clustering of the abundance profiles placed the paired specimens adjacent to each other. IHC H-score analysis of the validation set revealed a strong correlation between paired samples for all proteins. For the similarly expressed proteins, the estimated correlation coefficients in two of three experimental samples and all validation samples were statistically significant (p< 0.05). The estimated correlation coefficients in the experimental sample proteins classified as differentially expressed were not statistically significant.
  
  
  CONCLUSIONS
  A global proteomic screen of primary HGSOC tumors and their metastatic lesions identifies tumoral homogeneity and heterogeneity and provides preliminary insight into these protein profiles and the cellular pathways they constitute.
authors.affiliation: Division of Gynecologic Oncology, Magee-Womens Hospital of UPMC, Pittsburgh, PA 15213, USA. alsmith1@geisinger.edu.
authors.forename: Ashlee L
authors.initials: AL
authors.lastname: Smith
authors.affiliation: Biomedical Mass Spectrometry Center for the Health Sciences, University of Pittsburgh, Pittsburgh, PA 15213, USA. sunm3@upmc.edu.
authors.forename: Mai
authors.initials: M
authors.lastname: Sun
authors.affiliation: Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA. rbhargava@mail.magee.edu.
authors.forename: Rohit
authors.initials: R
authors.lastname: Bhargava
authors.affiliation: Biomedical Mass Spectrometry Center for the Health Sciences, University of Pittsburgh, Pittsburgh, PA 15213, USA. nas96@pitt.edu.
authors.forename: Nicolas A
authors.initials: NA
authors.lastname: Stewart
authors.affiliation: Departments of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA. cooperk3@upmc.edu.|Women's Cancer Research Center, Pittsburgh, PA 15213, USA. cooperk3@upmc.edu.|University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA. cooperk3@upmc.edu.
authors.forename: Melanie S
authors.initials: MS
authors.lastname: Flint
authors.affiliation: Women's Cancer Research Center, Pittsburgh, PA 15213, USA. bigbeewl@upmc.edu.
authors.forename: William L
authors.initials: WL
authors.lastname: Bigbee
authors.affiliation: Division of Gynecologic Oncology, Magee-Womens Hospital of UPMC, Pittsburgh, PA 15213, USA. tomkrivak@verizon.net.|Women's Cancer Research Center, Pittsburgh, PA 15213, USA. tomkrivak@verizon.net.|Magee-Womens Research Institute, Pittsburgh, PA 15213, USA. tomkrivak@verizon.net.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Krivak
authors.affiliation: Women's Cancer Research Center, Pittsburgh, PA 15213, USA. mstrange@mwri.magee.edu.
authors.forename: Mary A
authors.initials: MA
authors.lastname: Strange
authors.affiliation: University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA. cooperk3@upmc.edu.
authors.forename: Kristine L
authors.initials: KL
authors.lastname: Cooper
authors.affiliation: Division of Gynecologic Oncology, Magee-Womens Hospital of UPMC, Pittsburgh, PA 15213, USA. kkzorn@aol.com.|Women's Cancer Research Center, Pittsburgh, PA 15213, USA. kkzorn@aol.com.|Magee-Womens Research Institute, Pittsburgh, PA 15213, USA. kkzorn@aol.com.
authors.forename: Kristin K
authors.initials: KK
authors.lastname: Zorn
keywords: laser capture microdissection; proteomics; serous ovarian carcinoma