Increased WD-repeat containing protein 1 in interstitial fluid from ovarian carcinomas shown by comparative proteomic analysis of malignant and healthy gynecological tissue

Abstract

We aimed to identify differentially expressed proteins in interstitial fluid from ovarian cancer employing multiple fractioning and high resolution mass spectrometry-based proteomic analysis, and asked whether specific proteins that may serve as biomarker candidates or therapeutic targets could be identified. High throughput proteomics was conducted on immunodepleted and fractioned interstitial fluid from pooled samples of ovarian carcinomas, using endometrial carcinomas and healthy ovarian tissue as controls. Differential analysis revealed the up-regulation of extracellular proteasomes in tumor interstitial fluid compared to the healthy control. Moreover, a number of differentially expressed proteins in interstitial fluid from ovarian carcinomas compared with control tissues were identified. Detection of proteasome 20S related proteins in TIF compared to IF from healthy tissue indicates that the 20S proteasome can have a role in the tumor microenvironment. Six selected proteins, CEACAM5, FREM2, MUC5AC, TFF3, PYCARD and WDR1, were independently validated in individual tumor lysates from ovarian carcinomas by multiple reaction monitoring initiated detection and sequence analysis, Western blot and/or selected reaction monitoring. Quantification of specific proteins revealed substantial heterogeneity between individual samples. Nevertheless, WD repeat-containing protein 1 was confirmed as being significantly overexpressed in interstitial fluid from ovarian carcinomas compared to healthy ovarian tissue by Orbitrap analysis of individual native interstitial fluid from ovarian and endometrial carcinomas and healthy ovarian tissue. We suggest that this protein should be explored as a therapeutic target in ovarian carcinomas. This article is part of a Special Issue entitled: An Updated Secretome.

Keywords: - carcinoembryonic antigen-related adhesion molecule 5 (P06731); Biomarker; CEACAM5; EC; Endometrial cancer; FRAS1-related extracellular matrix protein 2 (Q5SZK8); FREM2; IF; Interstitial fluid; KEGG; Kyoto Encyclopedia of Genes and Genomes; MIDAS; MIF; MRM/SRM; MUC5AC; Microenvironment; OC; OH; Ovarian cancer; PYCARD; Proteomics; SIS; TBS; TFF3; TIF; WD repeat-containing protein 1 (O75083); WDR1; apoptosis-associated speck-like protein containing a CARD (Q9ULZ3); endometrial carcinoma (endometrial tumor tissue); interstitial fluid; mRP; macrophage migration inhibitor factor (P14174); macroporous reversed-phase chromatography; mucin 5AC (P98088); multiple reaction monitoring initiated detection and sequence analysis; multiple reaction monitoring/selected reaction monitoring; ovarian carcinoma (ovarian tumor tissue); ovarian healthy (healthy ovarian tissue); stable isotope-labeled internal standard; trefoil factor 3 (Q07654); tris-buffered saline; tumor interstitial fluid.