title: Cross-talk between non-genomic and genomic signalling pathways--distinct effect profiles of environmental estrogens.
abstract: Estrogen receptor (ER) transcriptional cross-talk after activation by 17beta-estradiol (E2) has been studied in considerable detail, but comparatively little is known about the ways in which synthetic estrogen-like chemicals, so-called xenoestrogens, interfere with these signalling pathways. E2 can stimulate rapid, non-genomic signalling events, such as activation of the Src/Ras/Erk signalling pathway. We investigated how activation of this pathway by E2, the estrogenic environmental contaminants o,p'-DDT, beta-HCH and p,p'-DDE, and epidermal growth factor (EGF) influences the expression of ER target genes, such as TFF1, ER, PR, BRCA1 and CCND1, and the proliferation of breast cancer cells. Despite commonalities in their estrogenicity as judged by cell proliferation assays, the environmental contaminants exhibited striking differences in their non-genomic and genomic signalling. The gene expression profiles of o,p'-DDT and beta-HCH resembled the effects observed with E2. In the case of beta-HCH this is surprising, considering its reported lack of affinity to the "classical" ER. The expression profiles seen with p,p'-DDE showed some similarities with E2, but overall, p,p'-DDE was a fairly weak transcriptional inducer of TFF1, ER, PR, BRCA1 and CCND1. We observed distinct differences in the non-genomic signalling of the tested compounds. p,p'-DDE was unable to stimulate Src and Erk1/Erk2 activations. The effects of E2 on Src and Erk1/Erk2 phosphorylation were transient and weak when compared to EGF, but beta-HCH induced strong and sustained activation of all tested kinases. Transcription of TFF1, ER, PR and BRCA1 by E2, o,p'-DDT and beta-HCH could be suppressed partially by inhibiting the Src/Ras/Erk pathway with PD 98059. However, this was not seen with p,p'-DDE. Our investigations show that the cellular activities of estrogens and xenoestrogens are the result of a combination of extranuclear (non-genomic) and nuclear (genomic) events and highlight the need to take non-genomic effects and signalling cross-talk into consideration, when screening for environmental estrogens. Otherwise, chemicals devoid of ER affinity, such as beta-HCH, but with an effect profile otherwise similar to estrogens might be overlooked in safety testing.
authors.affiliation: Centre for Toxicology, School of Pharmacy, University of London, 29-39 Brunswick Square, London, WC1N 1AX, UK. elisabete.silva@pharmacy.ac.uk
authors.forename: Elisabete
authors.initials: E
authors.lastname: Silva
authors.forename: Alena
authors.initials: A
authors.lastname: Kabil
authors.forename: Andreas
authors.initials: A
authors.lastname: Kortenkamp
mesh_terms: D019313:BRCA1 Protein / Q000096:biosynthesis / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019938:Cyclin D1 / Q000096:biosynthesis / Q000235:genetics; D003634:DDT / Q000633:toxicity; D003633:Dichlorodiphenyl Dichloroethylene / Q000494:pharmacology; D004785:Environmental Pollutants / Q000633:toxicity*; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000502:physiology; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology; D004965:Estrogen Antagonists / Q000494:pharmacology; D004967:Estrogens / Q000633:toxicity*; D005260:Female; D000077267:Fulvestrant; D020869:Gene Expression Profiling; D001556:Hexachlorocyclohexane / Q000494:pharmacology; D006801:Humans; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D011960:Receptors, Estrogen / Q000096:biosynthesis / Q000235:genetics; D015398:Signal Transduction; D000071163:Trefoil Factor-1; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis / Q000235:genetics; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D019313:BRCA1 Protein; C530104:CCND1 protein, human; D004785:Environmental Pollutants; D004965:Estrogen Antagonists; D004967:Estrogens; D011960:Receptors, Estrogen; C051838:TFF1 protein, human; D000071163:Trefoil Factor-1; D025521:Tumor Suppressor Proteins; D019938:Cyclin D1; D000077267:Fulvestrant; D003633:Dichlorodiphenyl Dichloroethylene; D004958:Estradiol; D001556:Hexachlorocyclohexane; D004815:Epidermal Growth Factor; D003634:DDT; C016340:o,p'-DDT; D019061:src-Family Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; C023888:beta-hexachlorocyclohexane