title: Protein profiles of cardiomyocyte differentiation in murine embryonic stem cells exposed to perfluorooctane sulfonate.
abstract: Perfluorooctane sulfonate (PFOS) is a persistent organic contaminant that may affect diverse systems in animals and humans, including the cardiovascular system. However, little is known about the mechanism by which it affects the biological systems. Herein, we used embryonic stem cell test procedure as a tool to assess the developmental cardiotoxicity of PFOS. The differentially expressed proteins were identified by quantitative proteomics that combines the stable isotope labeling of amino acids with high-performance liquid chromatography-electrospray ionization tandem mass spectrometry. Results of the embryonic stem cell test procedure suggested that PFOS was a weak embryotoxic chemical. Nevertheless, a few marker proteins related to cardiovascular development (Brachyury, GATA4, MEF2C, α-actinin) were significantly reduced by exposure to PFOS. In total, 176 differential proteins were identified by proteomics analysis, of which 67 were upregulated and 109 were downregulated. Gene ontology annotation classified these proteins into 13 groups by molecular functions, 12 groups by cellular locations and 10 groups by biological processes. Most proteins were mainly relevant to either catalytic activity (25.6%), nucleus localization (28.9%) or to cellular component organization (19.8%). Pathway analysis revealed that 32 signaling pathways were affected, particularly these involved in metabolism. Changes in five proteins, including L-threonine dehydrogenase, X-ray repair cross-complementing 5, superoxide dismutase 2, and DNA methyltransferase 3b and 3a were confirmed by Western blotting, suggesting the reliability of the technique. These results revealed potential new targets of PFOS on the developmental cardiovascular system.
authors.affiliation: Institute of Pharmacology, Toxicology, and Biochemical Pharmaceutics, Zhejiang University, Hangzhou, China.
authors.forename: Ying-Ying
authors.initials: YY
authors.lastname: Zhang
authors.affiliation: Institute of Pharmacology, Toxicology, and Biochemical Pharmaceutics, Zhejiang University, Hangzhou, China.
authors.forename: Lei-Lei
authors.initials: LL
authors.lastname: Tang
authors.affiliation: Institute of Pharmacology, Toxicology, and Biochemical Pharmaceutics, Zhejiang University, Hangzhou, China.
authors.forename: Bei
authors.initials: B
authors.lastname: Zheng
authors.affiliation: Institute of Reproductive Biomedicine and the 2nd Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
authors.forename: Ren-Shan
authors.initials: RS
authors.lastname: Ge
authors.affiliation: Institute of Pharmacology, Toxicology, and Biochemical Pharmaceutics, Zhejiang University, Hangzhou, China.
authors.forename: Dan-Yan
authors.initials: DY
authors.lastname: Zhu
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D017738:Alkanesulfonic Acids / Q000633:toxicity*; D000818:Animals; D041702:BALB 3T3 Cells; D002454:Cell Differentiation / Q000187:drug effects*; D019295:Computational Biology; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics / Q000378:metabolism; D000090205:DNA Methyltransferase 3A; D015536:Down-Regulation; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects*; D005466:Fluorocarbons / Q000633:toxicity*; D063990:Gene Ontology; D005819:Genetic Markers; D000072200:Ku Autoantigen / Q000235:genetics / Q000378:metabolism; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects*; D040901:Proteomics; D015203:Reproducibility of Results; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D059467:Transcriptome; D015854:Up-Regulation; D000097584:DNA Methyltransferase 3B; D000099156:Superoxide Dismutase 2
chemical_list: D000429:Alcohol Oxidoreductases; D017738:Alkanesulfonic Acids; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; D005466:Fluorocarbons; D005819:Genetic Markers; D000072200:Ku Autoantigen; D013482:Superoxide Dismutase; D000097584:DNA Methyltransferase 3B; D000099156:Superoxide Dismutase 2; C000717967:DNMT3A protein, human; C076994:perfluorooctane sulfonic acid; C023159:L-threonine 3-dehydrogenase
keywords: cardiomyocyte; embryonic stem cell test (EST); embryotoxicity; perfluorooctane sulfonate (PFOS); quantitative proteomics