Assessment of xenoestrogenic exposure by a biomarker approach: application of the E-Screen bioassay to determine estrogenic response of serum extractsReportar como inadecuado

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Environmental Health

, 2:12

First Online: 15 October 2003Received: 25 April 2003Accepted: 15 October 2003DOI: 10.1186-1476-069X-2-12

Cite this article as: Rasmussen, T.H., Nielsen, F., Andersen, H.R. et al. Environ Health 2003 2: 12. doi:10.1186-1476-069X-2-12


BackgroundEpidemiological documentation of endocrine disruption is complicated by imprecise exposure assessment, especially when exposures are mixed. Even if the estrogenic activity of all compounds were known, the combined effect of possible additive and-or inhibiting interaction of xenoestrogens in a biological sample may be difficult to predict from chemical analysis of single compounds alone. Thus, analysis of mixtures allows evaluation of combined effects of chemicals each present at low concentrations.

MethodsWe have developed an optimized in vitro E-Screen test to assess the combined functional estrogenic response of human serum. The xenoestrogens in serum were separated from endogenous steroids and pharmaceuticals by solid-phase extraction followed by fractionation by high-performance liquid chromatography. After dissolution of the isolated fraction in ethanol-DMSO, the reconstituted extract was added with estrogen-depleted fetal calf serum to MCF-7 cells, the growth of which is stimulated by estrogen. After a 6-day incubation on a microwell plate, cell proliferation was assessed and compared with the effect of a 17-beta-estradiol standard.

Results and conclusionsTo determine the applicability of this approach, we assessed the estrogenicity of serum samples from 30 pregnant and 60 non-pregnant Danish women thought to be exposed only to low levels of endocrine disruptors. We also studied 211 serum samples from pregnant Faroese women, whose marine diet included whale blubber that contain a high concentration of persistent halogenated pollutants. The estrogenicity of the serum from Danish controls exceeded the background in 22.7 % of the cases, while the same was true for 68.1 % of the Faroese samples. The increased estrogenicity response did not correlate with the lipid-based concentrations of individual suspected endocrine disruptors in the Faroese samples. When added along with the estradiol standard, an indication of an enhanced estrogenic response was found in most cases. Thus, the in vitro estrogenicity response offers a promising and feasible approach for an aggregated exposure assessment for xenoestrogens in serum.

List of abbreviations used4-OH-CBhydroxylated metabolites of polychlorinated biphenyls


CT-FCScharcoal-dextran-treated fetal calf serum

DMEMDulbecco-s modified Eagle-s medium



FCSfetal calf serum

GCgas chromatography


HEPES4-2-hydroxyethyl-1-piperazineethanesulfonic acid

HPLChigh-performance liquid chromatography

ICIICI 182,780

MOECmaximum observed effect concentration

NPCnormal phase chromatography

PCBpolychlorinated biphenyl

PEproliferative effect

RPErelative proliferative effect

RPPrelative proliferative potency

SPEsolid-phase extraction

SRBsulforhodamine B

TCAtrichloroacetic acid

Tris base2-amino-2-hydroxymethyl-1,3-propanediol.

Electronic supplementary materialThe online version of this article doi:10.1186-1476-069X-2-12 contains supplementary material, which is available to authorized users.

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Autor: Thomas Høj Rasmussen - Flemming Nielsen - Helle Raun Andersen - Jesper Bo Nielsen - Pal Weihe - Philippe Grandjean


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