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Proteomic-based identification of haptoglobin-1 precursor as a novel circulating biomarker of ovarian cancer

Screening for specific biomarkers of early-stage detection of ovarian cancer is a major health priority due to the asymptomatic nature and poor survival characteristic of the disease. We utilised two-dimensional gel electrophoresis (2DE) to identify differentially expressed proteins in the serum of...

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Autores principales: Ahmed, N, Barker, G, Oliva, K T, Hoffmann, P, Riley, C, Reeve, S, Smith, A I, Kemp, B E, Quinn, M A, Rice, G E
Formato: Texto
Lenguaje:English
Publicado: Nature Publishing Group 2004
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2364749/
https://www.ncbi.nlm.nih.gov/pubmed/15199385
http://dx.doi.org/10.1038/sj.bjc.6601882
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author Ahmed, N
Barker, G
Oliva, K T
Hoffmann, P
Riley, C
Reeve, S
Smith, A I
Kemp, B E
Quinn, M A
Rice, G E
author_facet Ahmed, N
Barker, G
Oliva, K T
Hoffmann, P
Riley, C
Reeve, S
Smith, A I
Kemp, B E
Quinn, M A
Rice, G E
author_sort Ahmed, N
collection PubMed
description Screening for specific biomarkers of early-stage detection of ovarian cancer is a major health priority due to the asymptomatic nature and poor survival characteristic of the disease. We utilised two-dimensional gel electrophoresis (2DE) to identify differentially expressed proteins in the serum of ovarian cancer patients that may be useful as biomarkers of this disease. In this study, 38 ovarian cancer patients at different pathological grades (grade 1 (n=6), grade 2 (n=8) and grade 3 (n=24)) were compared to a control group of eight healthy women. Serum samples were treated with a mixture of Affigel-Blue and protein A (5 : 1) for 1 h to remove high abundance protein (e.g. immunoglobulin and albumin) and were displayed using 11 cm, pH 4–7 isoelectric focusing strips for the first dimension and 10% acrylamide gel electrophoresis for the second dimension. Protein spots were visualised by SYPRO-Ruby staining, imaged by FX-imager and compared and analysed by PDQuest software. A total of 24 serum proteins were differentially expressed in grade 1 (P<0.05), 31 in grade 2 (P<0.05) and 25 in grade 3 (P<0.05) ovarian cancer patients. Six of the protein spots that were significantly upregulated in all groups of ovarian cancer patients were identified by nano-electrospray quadrupole quadrupole time-of-flight mass spectrometry (n-ESIQ(q)TOFMS) and matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOFMS) as isoforms of haptoglobin-1 precursor (HAP1), a liver glycoprotein present in human serum. Further identification of the spots at different pathological grades was confirmed by Western blotting using monoclonal antibody against a haptoglobin epitope contained within HAP1. Immunohistochemical localisation of HAP1-like activity was present in malignant ovarian epithelium and stroma but strong immunostaining was present in blood vessels, areas with myxomatous stroma and vascular spaces. No tissue localisation of HAP1-like immunoreactivity was observed in normal ovarian surface epithelium. These data highlight the need to assess circulating concentration of HAP1 in the serum of ovarian cancer patients and evaluate its potential as a biomarker in the early diagnosis of ovarian cancer.
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spelling pubmed-23647492009-09-10 Proteomic-based identification of haptoglobin-1 precursor as a novel circulating biomarker of ovarian cancer Ahmed, N Barker, G Oliva, K T Hoffmann, P Riley, C Reeve, S Smith, A I Kemp, B E Quinn, M A Rice, G E Br J Cancer Molecular and Cellular Pathology Screening for specific biomarkers of early-stage detection of ovarian cancer is a major health priority due to the asymptomatic nature and poor survival characteristic of the disease. We utilised two-dimensional gel electrophoresis (2DE) to identify differentially expressed proteins in the serum of ovarian cancer patients that may be useful as biomarkers of this disease. In this study, 38 ovarian cancer patients at different pathological grades (grade 1 (n=6), grade 2 (n=8) and grade 3 (n=24)) were compared to a control group of eight healthy women. Serum samples were treated with a mixture of Affigel-Blue and protein A (5 : 1) for 1 h to remove high abundance protein (e.g. immunoglobulin and albumin) and were displayed using 11 cm, pH 4–7 isoelectric focusing strips for the first dimension and 10% acrylamide gel electrophoresis for the second dimension. Protein spots were visualised by SYPRO-Ruby staining, imaged by FX-imager and compared and analysed by PDQuest software. A total of 24 serum proteins were differentially expressed in grade 1 (P<0.05), 31 in grade 2 (P<0.05) and 25 in grade 3 (P<0.05) ovarian cancer patients. Six of the protein spots that were significantly upregulated in all groups of ovarian cancer patients were identified by nano-electrospray quadrupole quadrupole time-of-flight mass spectrometry (n-ESIQ(q)TOFMS) and matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOFMS) as isoforms of haptoglobin-1 precursor (HAP1), a liver glycoprotein present in human serum. Further identification of the spots at different pathological grades was confirmed by Western blotting using monoclonal antibody against a haptoglobin epitope contained within HAP1. Immunohistochemical localisation of HAP1-like activity was present in malignant ovarian epithelium and stroma but strong immunostaining was present in blood vessels, areas with myxomatous stroma and vascular spaces. No tissue localisation of HAP1-like immunoreactivity was observed in normal ovarian surface epithelium. These data highlight the need to assess circulating concentration of HAP1 in the serum of ovarian cancer patients and evaluate its potential as a biomarker in the early diagnosis of ovarian cancer. Nature Publishing Group 2004-07-05 2004-06-15 /pmc/articles/PMC2364749/ /pubmed/15199385 http://dx.doi.org/10.1038/sj.bjc.6601882 Text en Copyright © 2004 Cancer Research UK https://creativecommons.org/licenses/by/4.0/This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material.If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.
spellingShingle Molecular and Cellular Pathology
Ahmed, N
Barker, G
Oliva, K T
Hoffmann, P
Riley, C
Reeve, S
Smith, A I
Kemp, B E
Quinn, M A
Rice, G E
Proteomic-based identification of haptoglobin-1 precursor as a novel circulating biomarker of ovarian cancer
title Proteomic-based identification of haptoglobin-1 precursor as a novel circulating biomarker of ovarian cancer
title_full Proteomic-based identification of haptoglobin-1 precursor as a novel circulating biomarker of ovarian cancer
title_fullStr Proteomic-based identification of haptoglobin-1 precursor as a novel circulating biomarker of ovarian cancer
title_full_unstemmed Proteomic-based identification of haptoglobin-1 precursor as a novel circulating biomarker of ovarian cancer
title_short Proteomic-based identification of haptoglobin-1 precursor as a novel circulating biomarker of ovarian cancer
title_sort proteomic-based identification of haptoglobin-1 precursor as a novel circulating biomarker of ovarian cancer
topic Molecular and Cellular Pathology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2364749/
https://www.ncbi.nlm.nih.gov/pubmed/15199385
http://dx.doi.org/10.1038/sj.bjc.6601882
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