Abstract 4800: Quantitative detection of discriminatory proteins in tissue and serum of ovarian cancer patients by LC-MSE
Authors: Wegdam Wouter , Meijer Danielle , Kramer Gertjan , Langridge James , Moerland Perry D. , de Jong Shreyas M. , Vissers Johannes P. , Kenter Gemma G. , Buist Marrije R. , Aerts Hans M.F.G. 
Published: August 2012

Purpose: Using label-free UPLC-MSE quantification methods to identifying proteins as well as determining their abundance in serum and microdissected ovarian cancer tissue. Experimental procedures: Serum of six patients with a serous adenocarcinoma of the ovary (stage IIIB or higher) was collected before treatment. Matched control patients with a serous cystadenoma of the ovary were chosen as a control group. In addition, homogeneous regions of cells that exhibited uniform histology were isolated from cancer tissue by laser capture microdissection (LCM), normal ovarian epithelial cell samples and stroma samples were taken from matched controls. We subsequently employed label-free ultra-high pressure liquid chromatography tandem mass spectrometry (UPLC-MSE) to identify proteins and determine their absolute concentrations in serum and tissue lysates. Moderated t-tests were used to identify differentially expressed proteins between patient- and control-group. p-values were adjusted for multiple-testing using Benjamini-Hochberg false discovery rates and considered differentially expressed when &lt;0.05. Summary of the data: In the serum specimens, 13 differentially expressed proteins were identified by LC-MSE profiling. Proteins with different concentrations in patients versus control sera include abundant serum proteins such as apolipoprotein AI and transferrin, both exhibited a lower concentration in serum of cancer patients. Differential expression was also observed for apolipoproteins (APOA IV, APOA II and APOC III) and other proteins that have not been associated with ovarian cancer previously such as C9 and Afamin. In the tissue-lysates we identified 535 Proteins of which 332 could be reliably quantified. Of these 56 were differentially expressed proteins of which 12 had non-overlapping standard deviations. Literature searches confirmed a link between ovarian cancer and 6 of these proteins, suggesting a possible role in disease progression. Of special interest was the identification of prohibitin (PHB) and cofilin-1 (CFL1). Immunohistochemical detection of CFL1 in formalin-fixed paraffin-embedded tissues of our patient group showed heterogeneous expression in 50% of the serous ovarian carcinomas, whereas all the benign lesions were negative. These results again demonstrate the validity of the protein quantitation results obtained by MSE. Interestingly, no overlap was found between the discriminatory proteins in serum specimens and corresponding tissue lysates, illustrating the challenge to identify a single biomarker in the circulation reflecting ovary cancer. Conclusions: Our study revealed several protein changes in serum and tissue of patients suffering from serous adenocarcinoma of the ovary. Further investigation of these proteins is warranted to establish whether they could provide new insights into disease etiology and act as potential new markers.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4800. doi:1538-7445.AM2012-4800