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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    • "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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