title: Additive estrogenic effects of mixtures of frequently used UV filters on pS2-gene transcription in MCF-7 cells.
abstract: In order to protect consumers from ultraviolet (UV) radiation and enhance light stability of the product, three to eight UV filters are usually added to consumer sunscreen products. High lipophilicity of the UV filters has been shown to cause bioaccumulation in fish and humans, leading to environmental levels of UV filters that are similar to those of PCBs and DDT. In this paper, estrogen-regulated pS2 gene transcription in the human mammary tumor cell line MCF-7 was used as a measure of estrogenicity of four individual UV filters. Since humans are exposed to more than one UV filter at a time, an equipotent binary mixture of 2-hydroxy-4-methoxy-benzophenone (BP-3) and its metabolite 2,4-dihydroxy benzophenone (BP-1), as well as an equipotent multi-component mixture of BP-1, BP-3, octyl methoxy cinnamate (OMC) and 3-(4-methylbenzylidene) camphor (4-MBC), were also evaluated for their ability to induce pS2 gene transcription in order to examine additivity. An estrogen receptor-mediated mechanism of action was expected for all UV filters. Therefore, our null-hypothesis was that combined estrogenic responses, measured as increased pS2 gene transcription in MCF-7 cells after exposure to mixtures of UV filters, are additive, according to a concentration-addition model. Not all UV filters produced a full concentration-response curve within the concentration range tested (100 nM-1 microM). Therefore, instead of using EC50 values for comparison, the concentration at which each compound caused a 50% increase of basal pS2 gene transcription was defined as the C50 value for that compound and used to calculate relative potencies. For comparison, the EC50 value of a compound is the concentration at which the compound elicits an effect that is 50% of its maximal effect. Individual UV filters increased pS2 gene transcription concentration-dependently with C50 values of 0.12 microM, 0.5 microM, 1.9 microM, and 1.0 microM for BP-1, BP-3, 4-MBC and OMC, respectively. Estradiol (E2) had a C50 value of 4.8 pM. Experiments with equipotent mixtures all supported our null hypothesis that mixtures of UV filters act additively to activate the estrogen receptor (ER). In view of our results and observed plasma levels it cannot be excluded that daily exposure to sunscreen formulations may have estrogenic effects in humans.
authors.affiliation: Institute for Risk Assessment Sciences, Utrecht University, PO Box 80176, 3508 TD Utrecht, The Netherlands. M.Heneweer@iras.uu.nl
authors.forename: Marjoke
authors.initials: M
authors.lastname: Heneweer
authors.forename: Martine
authors.initials: M
authors.lastname: Muusse
authors.forename: Martin
authors.initials: M
authors.lastname: van den Berg
authors.forename: J Thomas
authors.initials: JT
authors.lastname: Sanderson
mesh_terms: D000465:Algorithms; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D004338:Drug Combinations; D004347:Drug Interactions; D004958:Estradiol / Q000494:pharmacology; D045166:Estradiol Congeners; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D053766:Presenilin-2; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013473:Sunscreening Agents / Q000633:toxicity*; D014466:Ultraviolet Rays
chemical_list: D004338:Drug Combinations; D045166:Estradiol Congeners; D008565:Membrane Proteins; C508861:PSEN2 protein, human; D053766:Presenilin-2; D013473:Sunscreening Agents; D004958:Estradiol; D012313:RNA