Kristin L M Boylan1,2, Somaieh Afiuni-Zadeh1,3, Melissa A Geller4, Peter A Argenta4, Timothy J Griffin5, Amy P N Skubitz6,7,8. 1. Department of Laboratory Medicine & Pathology, University of Minnesota Medical School, MMC 395, 420 Delaware St. SE, Minneapolis, MN, 55455, USA. 2. Ovarian Cancer Early Detection Program, University of Minnesota Medical School, Minneapolis, MN, USA. 3. Lunenfeld-Tanenbaum Research Institute, Mount Sinai Health, Toronto, ON, M5G 1X5, Canada. 4. Department of Obstetrics, Gynecology, & Women's Health, University of Minnesota Medical School, Minneapolis, MN, USA. 5. Department of Biochemistry, Molecular Biology, & Biophysics, University of Minnesota Medical School, Minneapolis, MN, USA. 6. Department of Laboratory Medicine & Pathology, University of Minnesota Medical School, MMC 395, 420 Delaware St. SE, Minneapolis, MN, 55455, USA. skubi002@umn.edu. 7. Department of Obstetrics, Gynecology, & Women's Health, University of Minnesota Medical School, Minneapolis, MN, USA. skubi002@umn.edu. 8. Ovarian Cancer Early Detection Program, University of Minnesota Medical School, Minneapolis, MN, USA. skubi002@umn.edu.
Abstract
BACKGROUND: The purpose of this study was to determine whether the residual fixative from a liquid-based Pap test or a swab of the cervix contained proteins that were also found in the primary tumor of a woman with high grade serous ovarian cancer. This study is the first step in determining the feasibility of using the liquid-based Pap test or a cervical swab for the detection of ovarian cancer protein biomarkers. METHODS: Proteins were concentrated by acetone precipitation from the cell-free supernatant of the liquid-based Pap test fixative or eluted from the cervical swab. Protein was also extracted from the patient's tumor tissue. The protein samples were digested into peptides with trypsin, then the peptides were run on 2D-liquid chromatography mass spectrometry (2D-LCMS). The data was searched against a human protein database for the identification of peptides and proteins in each biospecimen. The proteins that were identified were classified for cellular localization and molecular function by bioinformatics integration. RESULTS: We identified almost 5000 proteins total in the three matched biospecimens. More than 2000 proteins were expressed in each of the three biospecimens, including several known ovarian cancer biomarkers such as CA125, HE4, and mesothelin. By Scaffold analysis of the protein Gene Ontology categories and functional analysis using PANTHER, the proteins were classified by cellular localization and molecular function, demonstrating that the Pap test fluid and cervical swab proteins are similar to each other, and also to the tumor extract. CONCLUSIONS: Our results suggest that Pap test fixatives and cervical swabs are a rich source of tumor-specific biomarkers for ovarian cancer, which could be developed as a test for ovarian cancer detection.
BACKGROUND: The purpose of this study was to determine whether the residual fixative from a liquid-based Pap test or a swab of the cervix contained proteins that were also found in the primary tumor of a woman with high grade serous ovarian cancer. This study is the first step in determining the feasibility of using the liquid-based Pap test or a cervical swab for the detection of ovarian cancer protein biomarkers. METHODS: Proteins were concentrated by acetone precipitation from the cell-free supernatant of the liquid-based Pap test fixative or eluted from the cervical swab. Protein was also extracted from the patient's tumor tissue. The protein samples were digested into peptides with trypsin, then the peptides were run on 2D-liquid chromatography mass spectrometry (2D-LCMS). The data was searched against a human protein database for the identification of peptides and proteins in each biospecimen. The proteins that were identified were classified for cellular localization and molecular function by bioinformatics integration. RESULTS: We identified almost 5000 proteins total in the three matched biospecimens. More than 2000 proteins were expressed in each of the three biospecimens, including several known ovarian cancer biomarkers such as CA125, HE4, and mesothelin. By Scaffold analysis of the protein Gene Ontology categories and functional analysis using PANTHER, the proteins were classified by cellular localization and molecular function, demonstrating that the Pap test fluid and cervical swab proteins are similar to each other, and also to the tumor extract. CONCLUSIONS: Our results suggest that Pap test fixatives and cervical swabs are a rich source of tumor-specific biomarkers for ovarian cancer, which could be developed as a test for ovarian cancer detection.
Entities:
Keywords:
Biomarker; Mass spectrometry based proteomics; Ovarian cancer detection; Pap test
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