Atrazine and PCB 153 and their effects on the proteome of subcellular fractions of human MCF-7 cells

Centre de Recherche Public - Gabriel Lippmann, Department of Environment and Agro-Biotechnologies, 41, rue du Brill, L-4422 Belvaux, Luxembourg.
Biochimica et Biophysica Acta (Impact Factor: 4.66). 04/2012; 1824(6):833-41. DOI: 10.1016/j.bbapap.2012.03.014
Source: PubMed


Several man-made organic pollutants including polychlorinated biphenyls (PCBs) and several pesticides may exhibit endocrine disrupting (ED) properties. These ED molecules can be comparatively persistent in the environment, and have shown to perturb hormonal activity and several physiological functions. The objective of this investigation was to study the impact of PCB 153 and atrazine on human MCF-7 cells, and to search for marker proteins of their exposure. Cells were exposed to environmentally high but relevant concentrations of atrazine (200ppb), PCB 153 (500ppb), 17-β estradiol (positive control, 10nM) and DMSO (0.1%, negative control) for t=36h (n=3 replicates/exposure group). Proteins from cell membrane and cytosol were isolated, and studied by 2D-DiGE. Differentially regulated proteins were trypsin-digested and identified by MALDI-ToF-ToF and NCBInr database. A total of 36 differentially regulated proteins (>|1.5| fold change, P<0.05) were identified in the membrane fraction and 22 in the cytosol, and were mainly involved in cell structure and in stress response, but also in xenobiotic metabolism. 67% (membrane) and 50% (cytosol) of differentially regulated proteins were more abundant following atrazine exposure whereas nearly 100% (membrane) and 45% (cytosol) were less abundant following PCB 153 exposure. Western blots of selected proteins (HSBP1, FKBP4, STMN1) confirmed 2D-DiGE results. This study emphasizes the numerous potential effects that ED compounds could have on exposed humans.

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Available from: Torsten Bohn, Oct 07, 2015
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    • "Atrazine (2-chloro-4-[ethylamino]-6-[isopropylamino]-s-triaz- ine) herbicide belongs to the triazine family, and has been widely used to control weeds in crops. Atrazine is highly mobile toxic to aquatic organisms, plants, and human beings (Lasserre et al., 2012). Techniques for removing triazine from aqueous solutions 0960-8524/Ó 2015 Elsevier Ltd. "
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    • "). Prefractionation of the original sample is an appropriate means to arrive at subproteomes, for example, of subcellular organelles (Bruno et al., 2002; Lasserre et al., 2012). In addition, chromatographic steps (1) based on different separation criteria and (2) taking advantage of specific protein properties, such as isoelectric point, hydrophobicity , and interaction with affinity matrices, may reduce sample complexity and reveal otherwise hidden proteins (Miller, 2011; Capriotti et al., 2012). "
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    ABSTRACT: Polychlorinated biphenyls (PCB), synthetic, persistent organic pollutants (POP), are detected ubiquitously, in water, soil, air, and sediments, as well as in animals and humans. PCB are associated with range of adverse health effects, such as interference with the immune system and nervous system, reproductive abnormalities, fetotoxicity, carcinogenicity, and endocrine disruption. Our objective was to determine the effects of three structurally different PCB congeners, PCB118, PCB 126, and PCB 153, each at two concentrations, on the steroidogenic capacity and proteome of human adrenocortical carcinoma cell line cultures (H295R) . After 48 h of exposure, cell viability was monitored and estradiol, testosterone, cortisol and progesterone secretion measured to quantify steroidogenic capacity of the cells. Two-dimensional (2D) gel-based proteomics was used to screen for proteome alterations in H295R cells in response to the PCB. Exposure to PCB 118 increased estradiol and cortisol secretion, while exposure to PCB 153 elevated estradiol secretion. PCB 126 was the most potent congener, increasing estradiol, cortisol, and progesterone secretion in exposed H295R cells. Seventy-three of the 711 spots analyzed showed a significant difference in normalized spot volumes between controls (vehicle only) and at least one exposure group. Fourteen of these protein spots were identified by liquid chromatography with mass spectroscopy (LC-MS/MS). Exposure to three PCB congeners with different chemical structure perturbed steroidogenesis and protein expression in the H295R in vitro model. This study represents an initial analysis of the effects on proteins and hormones in the H295R cell model, and additional studies are required in order to obtain a more complete understanding of the pathways disturbed by PCB congeners in H295R cells. Overall, alterations in protein regulation and steroid hormone synthesis suggest that exposure to PCB disturbs several cellular processes, including protein synthesis, stress response, and apoptosis.
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