Two-Dimensional Differential Gel Electrophoresis to Identify Protein Biomarkers in Amniotic Fluid of Edwards Syndrome (Trisomy 18) Pregnancies |
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Authors: | Te-Yao Hsu Hao Lin Hsuan-Ning Hung Kuender D Yang Chia-Yu Ou Ching-Chang Tsai Hsin-Hsin Cheng Su-Hai Chung Bi-Hua Cheng Yi-Hsun Wong An Kuo Chou Chang-Chun Hsiao |
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Institution: | 1. Department of Obstetrics and Gynecology, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan;2. Department of Pediatrics, Chang Bing Show Chwan Memorial Hospital, Chang Hwa, Taiwan;3. Department of Anesthesia, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan;4. Genomic Medicine Research Core Laboratory, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan;5. Chang Gung University College of Medicine, Kaohsiung, Taiwan;Universidad Nacional de la Plata, ARGENTINA |
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Abstract: | BackgroundEdwards syndrome (ES) is a severe chromosomal abnormality with a prevalence of about 0.8 in 10,000 infants born alive. The aims of this study were to identify candidate proteins associated with ES pregnancies from amniotic fluid supernatant (AFS) using proteomics, and to explore the role of biological networks in the pathophysiology of ES.MethodsAFS from six second trimester pregnancies with ES fetuses and six normal cases were included in this study. Fluorescence-based two-dimensional difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) were used for comparative proteomic analysis. The identified proteins were further validated by Western blotting and the role of biological networks was analyzed.ResultsTwelve protein spots were differentially expressed by more than 1.5-fold in the AFS of the ES pregnancies. MALDI-TOF/MS identified one up-regulated protein: apolipoprotein A1 (ApoA1), and four under-regulated proteins: vitamin D binding protein (VDBP), alpha-1-antitrypsin (A1AT), insulin-like growth factor-binding protein 1 (IGFBP-1), and transthyretin (TTR). Western blot and densitometric analysis of ApoA1, A1AT, IGFBP-1, and TTR confirmed the alteration of these proteins in the amniotic fluid samples. Biological network analysis revealed that the proteins of the ES AFS were involved mainly in lipid and hormone metabolism, immune response, and cardiovascular disease.ConclusionsThese five proteins may be involved in the pathogenesis of ES. Further studies are needed to explore. |
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