Abstract

Objective: The biology of high-grade serous ovarian carcinoma (HGSOC) is poorly understood. Access to fresh-frozen primary tumors and matched metastatic sites is limited, but formalin-fixed, paraffin-embedded (FFPE) tissues are more readily available. Proteomic analysis of HGSOC thus far has focused on the identification of serum biomarkers, while little data have been reported regarding heterogeneity of primary ovarian tumors and their metastases. In this pilot study, our goal was to evaluate the protein expression profiles of paired primary and metastatic tumor from FFPE ovarian samples.

Highlights

  • Epithelial ovarian cancer (EOC) is the most lethal gynecologic malignancy [1]

  • Scans performed in the linear ion trap (LIT) where the seven most abundant peptide molecular ions were selected for collision-induced dissociation (CID), using a normalized collision energy of 35%

  • E-HGSOC: experimental high grade serous ovarian carcinoma samples; V-HGSOC: validation high grade serous ovarian carcinoma samples; The H-score is given as the sum of the percent staining multiplied by an ordinal value corresponding to the intensity level (0 = none, 1 = weak, 2 = moderate, 3 = strong)

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Summary

Introduction

The majority of patients with EOC present with metastatic lesions identified at primary surgery and experience recurrence This type of clinical behavior affords researchers the opportunity to evaluate multiple tumor samples throughout a patient’s clinical course. Attempts to identify serum biomarkers, investigations of the histologic types, and genomic studies of metastatic lesions have been completed utilizing fresh-frozen samples [5,6,7,8,9]. Proteomic studies utilized HGSOC tissue sources ranging from fresh-frozen specimens to ovarian cancer cell lines [13,22]. Our goal was to evaluate the protein expression profiles via a global proteomic screening of HGSOC from FFPE samples of both ovaries, with the presumption that one side represents the primary tumor and the other side a metastatic site, eliminating heterogeneity among different tissue types. Following identification, selected proteins involved with migration, cell adhesion, and metastasis were investigated using IHC to assess the correlation of findings

Experimental
Liquid Chromatography Tandem Mass Spectrometry
Peptide Identification
Spectral Count Analysis
Ingenuity Pathways Analysis
Immunohistochemical Staining
Results
Discussion
Conclusions
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