title: Combination of Metabolomics with Cellular Assays Reveals New Biomarkers and Mechanistic Insights on Xenoestrogenic Exposures in MCF-7 Cells.
abstract: The disruptive potential of xenoestrogens like bisphenol A (BPA) lies in their 17β-estradiol (E2)-like binding to estrogen receptors (ERs) followed by concomitant modulation of ER target gene expression. Unsurprisingly, most endocrine testing systems focus on the quantification of canonical transcripts or ER-sensitive reporters. However, only little information is available about the corresponding metabolomic changes in vitro. This knowledge gap becomes particularly relevant in the context of potential mixture effects, for example, as a consequence of coexposure to potentially estrogenically active pollutants (e.g., Cd2+). Such effects are often difficult to dissect with molecular tools, especially with regard to potential physiological relevance. Metabolomic biomarkers are well-suited to address this latter aspect as they provide a comprehensive readout of whole-cell physiology. Applying a targeted metabolomics approach (FIA-MS/MS), this study looked for biomarkers indicative of xenoestrogenic exposure in MCF-7 cells. Cells were treated with E2 and BPA in the presence or absence of Cd2+. Statistical analysis revealed a total of 11 amino acids and phospholipids to be related to the compound's estrogenic potency. Co-exposure to Cd2+modulated the estrogenic profile. However, the corresponding changes were found to be moderate with cellular assays such as the E-screen failing to record any Cd2+-specific estrogenic effects. Overall, metabolomics analysis identified proline as the most prominent estrogenic biomarker. Its increase could clearly be related to estrogenic exposure and concomitant ERα-mediated induction of proliferation. Involvement of the latter was confirmed by siRNA-mediated knockdown studies as well as by receptor inhibition. Further, the underlying signaling was also found to involve the oncoprotein MYC. Taken together, this study provides insights into the underlying mechanisms of xenoestrogenic effects and exemplify the strength of the complementary use of metabolomics and cellular and molecular assays.
authors: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR) , Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors: Sarah
authors: S
authors: Potratz
authors: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR) , Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors: Patrick
authors: P
authors: Tarnow
authors: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR) , Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors: Harald
authors: H
authors: Jungnickel
authors: Department of Molecular Systems Biology, UFZ-Helmholtz-Centre for Environmental Research , Permoserstrasse 15, 04318 Leipzig, Germany.|Institute of Pharmacy, Faculty of Biosciences, Pharmacology and Psychology, University of Leipzig , Brüderstrasse 34, 04103 Leipzig, Germany.
authors: Sven
authors: S
authors: Baumann
authors: Department of Molecular Systems Biology, UFZ-Helmholtz-Centre for Environmental Research , Permoserstrasse 15, 04318 Leipzig, Germany.|Institute of Biochemistry, Faculty of Biosciences, Pharmacology and Psychology, University of Leipzig , Brüderstrasse 34, 04103 Leipzig, Germany.|Department of Biotechnology, Chemistry and Environmental Engineering, Aalborg University , Fredrik Bajers Vej 7K, 9220 Aalborg, Denmark.
authors: Martin
authors: M
authors: von Bergen
authors: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR) , Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors: Tewes
authors: T
authors: Tralau
authors: Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR) , Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors: Andreas
authors: A
authors: Luch
mesh_terms: D001559:Benzhydryl Compounds / Q000737:chemistry / Q000633:toxicity; D015415:Biomarkers / Q000378:metabolism*; D002104:Cadmium / Q000737:chemistry; D049109:Cell Proliferation / Q000187:drug effects*; D003124:Colorimetry; D016002:Discriminant Analysis; D052244:Endocrine Disruptors / Q000737:chemistry / Q000633:toxicity*; D004958:Estradiol / Q000737:chemistry / Q000633:toxicity; D047628:Estrogen Receptor alpha / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter; D006801:Humans; D061986:MCF-7 Cells; D055442:Metabolome / Q000187:drug effects; D055432:Metabolomics; D010636:Phenols / Q000737:chemistry / Q000633:toxicity; D011392:Proline / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D000098826:Bisphenol A Compounds
chemical_list: D001559:Benzhydryl Compounds; D015415:Biomarkers; D002104:Cadmium; D052244:Endocrine Disruptors; D004958:Estradiol; D047628:Estrogen Receptor alpha; D010636:Phenols; D011392:Proline; D016271:Proto-Oncogene Proteins c-myc; D034741:RNA, Small Interfering; C506487:ESR1 protein, human; C006780:bisphenol A; D000098826:Bisphenol A Compounds