Development of a stably transfected estrogen receptor-mediated luciferase reporter gene assay in the human T47D breast cancer cell line.

Development of an estrogen receptor-mediated, chemical-activated luciferase reporter gene-expression (ER-CALUX) assay was attempted by stable transfection of luciferase reporter genes in a number of cell lines. Stable transfection of the chimeric Gal4 estrogen receptor and luciferase gene constructs in MCF-7 breast cancer and Hepa.1c1c7 mouse hepatoma cell lines, as well as transfection of a newly constructed luciferase reporter gene pEREtata-Luc in the ECC-1 human endometrial cell line, resulted in constitutive, non-estradiol-inducible clones. Stable transfection of pEREtata-Luc in the T47D breast cancer cell line, however, resulted in an extremely sensitive, highly responsive cell line. Following a 24-h exposure to estradiol (E2), stably transfected T47D.Luc cells demonstrated a detection limit of 0.5 pM, an EC50 of 6 pM, and a maximum induction of 100-fold relative to solvent controls. No clear reduction in responsiveness has been found over extended culture periods (50 passages). Anti-estrogens ICI 182,780, TCDD, and tamoxifen inhibited the estradiol-mediated luciferase induction. Genistein, nonylphenol, and o,p'DDT were the most potent (pseudo-)estrogens tested in this system (EC50 100, 260, and 660 nM, respectively). Determination of interactive effects of the (pseudo-)estrogens nonylphenol, o,p'DDT, chlordane, endosulfan, dieldrin, and methoxychlor revealed that, in combination with 3 pM E2, (pseudo-)estrogens were additive. Slightly more than additive effects (less than 2-fold) were found for combinations of dieldrin and endosulfan tested in the range of 3 to 6 microM. At these concentrations, the combination of endosulfan and chlordane demonstrated additive interaction. The ER-CALUX assay with T47D cells can provide a sensitive, responsive, and rapid in vitro system to detect and measure substances with potential (anti-)estrogenic activity.

[1]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[2]  J. Corton,et al.  Interaction of Estrogenic Chemicals and Phytoestrogens with Estrogen Receptor β. , 1998, Endocrinology.

[3]  B. van der Burg,et al.  Interference between Progesterone and Dioxin Signal Transduction Pathways , 1998, The Journal of Biological Chemistry.

[4]  T. Sutter,et al.  Differential expression of CYP1A1 and CYP1B1 in human breast epithelial cells and breast tumor cells. , 1998, Carcinogenesis.

[5]  K. Korach,et al.  A role for oestrogens in the male reproductive system , 1997, Nature.

[6]  D. Schlenk,et al.  Influence of β-Naphthoflavone and Methoxychlor Pretreatment on the Biotransformation and Estrogenic Activity of Methoxychlor in Channel Catfish (Ictalurus punctatus)☆ , 1997 .

[7]  Helmut Dotzlaw,et al.  Expression of estrogen receptor-beta in human breast tumors. , 1997, The Journal of clinical endocrinology and metabolism.

[8]  J. Giesy,et al.  Hydroxylated polychlorinated biphenyl metabolites are anti-estrogenic in a stably transfected human breast adenocarcinoma (MCF7) cell line. , 1997, Toxicology and applied pharmacology.

[9]  T. Zacharewski.  In Vitro Bioassays for Assessing Estrogenic Substances , 1997 .

[10]  M D Shelby,et al.  Assessing environmental chemicals for estrogenicity using a combination of in vitro and in vivo assays. , 1996, Environmental Health Perspectives.

[11]  S. Hill,et al.  Identification of environmental chemicals with estrogenic activity using a combination of in vitro assays. , 1996, Environmental health perspectives.

[12]  J. Giesy,et al.  Chemical-activated luciferase gene expression (CALUX): a novel in vitro bioassay for Ah receptor active compounds in sediments and pore water. , 1996, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[13]  J. Polman,et al.  ERβ: Identification and characterization of a novel human estrogen receptor , 1996 .

[14]  J. Gustafsson,et al.  Cloning of a novel receptor expressed in rat prostate and ovary. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[15]  J. McLachlan,et al.  A yeast estrogen screen for examining the relative exposure of cells to natural and xenoestrogens. , 1996, Environmental health perspectives.

[16]  J. Giesy,et al.  Species-specific recombinant cell lines as bioassay systems for the detection of 2,3,7,8-tetrachlorodibenzo-p-dioxin-like chemicals. , 1996, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[17]  Edwin J. Routledge,et al.  Estrogenic activity of surfactants and some of their degradation products assessed using a recombinant yeast screen , 1996 .

[18]  T. Zacharewski,et al.  Assessing the estrogenic and dioxin-like activities of chemicals and complex mixtures using in vitro recombinant receptor-reporter gene assays. , 1996, Canadian journal of physiology and pharmacology.

[19]  M. Schalk,et al.  Species-specific antagonism of Ah receptor action by 2,2',5,5'-tetrachloro- and 2,2',3,3'4,4'-hexachlorobiphenyl. , 1995, European journal of pharmacology.

[20]  C Sonnenschein,et al.  The E-SCREEN assay as a tool to identify estrogens: an update on estrogenic environmental pollutants. , 1995, Environmental health perspectives.

[21]  R. Dickson,et al.  Growth factors in breast cancer. , 1995, Endocrine reviews.

[22]  T. Zacharewski,et al.  Detection of Estrogen- and Dioxin-like Activity in Pulp and Paper Mill Black Liquor and Effluent Using in vitro Recombinant Receptor/Reporter Gene Assays. , 1995, Environmental science & technology.

[23]  J. Sumpter,et al.  A variety of environmentally persistent chemicals, including some phthalate plasticizers, are weakly estrogenic. , 1995, Environmental health perspectives.

[24]  K. Korach,et al.  Anti-estrogen activity in the yeast transcription system: Estrogen receptor mediated agonist response , 1994, Steroids.

[25]  J. Sumpter,et al.  Environmentally persistent alkylphenolic compounds are estrogenic. , 1994, Endocrinology.

[26]  S. Gelmini,et al.  Chimeric receptors as a tool for luminescent measurement of biological activities of steroid hormones. , 1994, Journal of bioluminescence and chemiluminescence.

[27]  C. Sonnenschein,et al.  The pesticides endosulfan, toxaphene, and dieldrin have estrogenic effects on human estrogen-sensitive cells. , 1994, Environmental health perspectives.

[28]  John P. Sumpter,et al.  Detergent components in sewage effluent are weakly oestrogenic to fish: An in vitro study using rainbow trout (Oncorhynchus mykiss) hepatocytes , 1993 .

[29]  T. Zacharewski,et al.  ICI 164,384: a control for investigating estrogen responsive genes. , 1993, Nucleic acids research.

[30]  A. Soto,et al.  Developmental effects of endocrine-disrupting chemicals in wildlife and humans. , 1993, Environmental health perspectives.

[31]  U. Mayr,et al.  Validation of two in vitro test systems for estrogenic activities with zearalenone, phytoestrogens and cereal extracts. , 1992, Toxicology.

[32]  C. Lyttle,et al.  Human estrogen receptor regulation in a yeast model system and studies on receptor agonists and antagonists , 1992, The Journal of Steroid Biochemistry and Molecular Biology.

[33]  D. Röhme,et al.  Different pathways for chromosomal integration of transfected circular pSVneo plasmids in normal and established rodent cells. , 1992, Gene.

[34]  K. Korach,et al.  Estrogens modulate the responsiveness of osteoblast-like cells (ROS 17/2.8) stably transfected with estrogen receptor. , 1992, Endocrinology.

[35]  C. Sonnenschein,et al.  p-Nonyl-phenol: an estrogenic xenobiotic released from "modified" polystyrene. , 1991, Environmental health perspectives.

[36]  A. Berns,et al.  Consecutive inactivation of both alleles of the pim-1 proto-oncogene by homologous recombination in embryonic stem cells , 1990, Nature.

[37]  D Gagne,et al.  A new cellular model of response to estrogens: a bioluminescent test to characterize (anti) estrogen molecules. , 1990, BioTechniques.

[38]  Toru Watanabe,et al.  In vitro Sensitivity Test of Breast Cancer Cells to Hormonal Agents in a Radionucleotide‐incorporation Assay , 1990, Japanese journal of cancer research : Gann.

[39]  H. Rochefort,et al.  Regulation of cathepsin-D and pS2 gene expression by growth factors in MCF7 human breast cancer cells. , 1989, Molecular endocrinology.

[40]  R. Evans,et al.  The steroid and thyroid hormone receptor superfamily. , 1988, Science.

[41]  H. Rochefort,et al.  Estrogens and growth factors induce the mRNA of the 52K-pro-cathepsin-D secreted by breast cancer cells. , 1988, Nucleic acids research.

[42]  S. Green,et al.  A versatile in vivo and in vitro eukaryotic expression vector for protein engineering , 1988, Nucleic Acids Res..

[43]  M. Blankenstein,et al.  Mitogenic stimulation of human breast cancer cells in a growth factor‐defined medium: Synergistic action of insulin and estrogen , 1988, Journal of cellular physiology.

[44]  B. Katzenellenbogen,et al.  Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[45]  P. Goad,et al.  Prenatal chlordane exposure: effects on plasma corticosterone concentrations over the lifespan of mice. , 1984, Environmental research.

[46]  D. Fry,et al.  DDT-induced feminization of gull embryos. , 1981, Science.

[47]  W. McGuire,et al.  Estrogen control of progesterone receptor in human breast cancer. Correlation with nuclear processing of estrogen receptor. , 1978, The Journal of biological chemistry.

[48]  S. J. Harris,et al.  Estrogenic Activity of o, p'-DDT in the Mammalian Uterus and Avian Oviduct , 1968, Science.

[49]  J. Corton,et al.  Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor beta. , 1998, Endocrinology.

[50]  Ying Li,et al.  Differential expression of CYP 1 A 1 and CYP 1 B 1 in human breast epithelial cells and breast tumor cells , 1998 .

[51]  J. Gustafsson,et al.  Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors alpha and beta. , 1997, Endocrinology.

[52]  R. Vihko,et al.  Retinoic acids increase 17 beta-hydroxysteroid dehydrogenase type 1 expression in JEG-3 and T47D cells, but the stimulation is potentiated by epidermal growth factor, 12-O-tetradecanoylphorbol-13-acetate, and cyclic adenosine 3',5'-monophosphate only in JEG-3 cells. , 1997, Endocrinology.

[53]  K. Grandien,et al.  Comparison of the Ligand Binding Specificity and Transcript Tissue Distribution of Estrogen Receptors a and b , 1997 .

[54]  S. Safe,et al.  Modulation of gene expression and endocrine response pathways by 2,3,7,8-tetrachlorodibenzo-p-dioxin and related compounds. , 1995, Pharmacology & therapeutics.

[55]  B. Weintraub.  Molecular endocrinology : basic concepts and clinical correlations , 1995 .

[56]  K. Wood,et al.  Firefly Luciferase Engineered for Improved Genetic Reporting , 1994 .

[57]  L. Gray,et al.  A dose-response analysis of methoxychlor-induced alterations of reproductive development and function in the rat. , 1989, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[58]  L. Gray,et al.  A Dose-Response Analysis of Methoxychlor-Induced Alterations of Reproductive Development and Function in the Rat , 1989 .

[59]  S. Safe,et al.  The effects of organochlorine pesticides as inducers of testosterone and benzo[a]pyrene hydroxylases. , 1987, General pharmacology.

[60]  B. Burg,et al.  Mitogenic stimulation of human breast cancer cells in a growth factor-defined medium: Synergistic action of insulin and estrogens , 1987 .

[61]  M. Metzler,et al.  Estrogenic activity in vivo and in vitro of some diethylstilbestrol metabolites and analogs. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[62]  J. Gustafsson,et al.  Cloning of a novel estrogen receptor expressed in rat prostate and ovary , 2022 .

[63]  K. Grandien,et al.  Printed in U.S.A. Copyright © 1997 by The Endocrine Society Comparison of the Ligand Binding Specificity and Transcript Tissue Distribution of Estrogen Receptors � and � , 2022 .