TY - JOUR
T1 - Atrazine and PCB 153 and their effects on the proteome of subcellular fractions of human MCF-7 cells
AU - Lasserre, Jean Paul
AU - Fack, Fred
AU - Serchi, Tommaso
AU - Revets, Dominique
AU - Planchon, Sébastien
AU - Renaut, Jenny
AU - Hoffmann, Lucien
AU - Gutleb, Arno C.
AU - Muller, Claude P.
AU - Bohn, Torsten
N1 - Funding Information:
We are grateful to Dr. Hilda Witters who participated in the early phase of the project during the transfer of the MCF7/BUS cells to LNS II and gave guidelines for setting up of the initial culture and exposure methods. This work was supported by the FNR (Fonds National de la Recherche) project ENDIF ( FNR/03/07/05 ) within the “FOOD-SAFETY (SECAL)” program.
PY - 2012/6
Y1 - 2012/6
N2 - 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 (200 ppb), PCB 153 (500 ppb), 17-β estradiol (positive control, 10 nM) and DMSO (0.1%, negative control) for t = 36 h (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.
AB - 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 (200 ppb), PCB 153 (500 ppb), 17-β estradiol (positive control, 10 nM) and DMSO (0.1%, negative control) for t = 36 h (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.
KW - 2D-DiGE
KW - Atrazine
KW - MCF-7 human cell
KW - PCB 153
KW - Subcellular fraction
UR - http://www.scopus.com/inward/record.url?scp=84860333670&partnerID=8YFLogxK
U2 - 10.1016/j.bbapap.2012.03.014
DO - 10.1016/j.bbapap.2012.03.014
M3 - Article
C2 - 22516319
AN - SCOPUS:84860333670
SN - 1570-9639
VL - 1824
SP - 833
EP - 841
JO - Biochimica et Biophysica Acta - Proteins and Proteomics
JF - Biochimica et Biophysica Acta - Proteins and Proteomics
IS - 6
ER -