Affiliations: [a] Fundamental and Computational Sciences, Pacific Northwest National Laboratory, Richland, WA, USA | [b] Division of Radiological Sciences, Washington University School of Medicine, St. Louis, MO, USA | [c] Statistical and Mathematical Sciences, Pacific Northwest National Laboratory, Richland, WA, USA | [d] Department of Surgery, Duke University Medical Center, Durham, NC, USA | [x] Department of Breast medical Oncology, M.D. Anderson Cancer Center, Houston, TX, USA | [y] Department of Gynecologic Oncology, M.D. Anderson Cancer Center, Houston, TX, USA
Correspondence:
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Corresponding author: Richard C. Zangar, Fundamental {and} Computational Sciences, Pacific Northwest National Laboratory, Richland, P.O. Box 999, WA, USA. E-mail: [email protected].
Abstract: Background:Post-translational protein modifications (PTMs) are increased in breast tumors. Objective:We explored whether PTMs on proteins secreted by the breast could be detected in plasma and potentially used for the early detection of breast cancer. Methods:We used a custom ELISA microarray platform to measure 4-hydroxynonenal (HNE), glutathione (GSH), nitrotyrosine and halotyrosine adducts in 27 secreted proteins, for a total of 108 candidate biomarkers. Two independent sets of human plasma samples were measured, for a total of 160 samples. The results were analyzed for consistent cancer-associated changes across the two sample sets. Plasma samples for both cases and benign controls were collected at the time of tissue diagnosis after referral from a positive screen (such as mammography). The results from both studies were evaluated using ANOVA and t-tests or receiver operator curves (ROC). Results:Levels of GSH-modified ceruloplasmin and HNE-modified PDGF were significantly altered in plasma samples from cancer patients relative to benign controls. Healthy controls, which were only included in the first set of samples, were similar to the benign controls for both of these markers. A combination of three glutathionylated proteins produced the best area under the ROC curve, with a value of 76%. Conclusions:Specific PTMs in individual proteins may be useful for distinguishing between women with breast cancer and those with benign breast disease. These oxidative changes in plasma proteins may reflect redox changes in breast cancer. Additional studies on oxidative modifications in individual proteins are warranted.
Keywords: Breast cancer, biomarkers, reactive oxygen species, protein adducts, hydroxynonenal, glutathione, plasma
