Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Environ Health Perspect
2008 Feb 01;1162:165-72. doi: 10.1289/ehp.10131.
Show Gene links
Show Anatomy links
Arsenic as an endocrine disruptor: arsenic disrupts retinoic acid receptor-and thyroid hormone receptor-mediated gene regulation and thyroid hormone-mediated amphibian tail metamorphosis.
Davey JC
,
Nomikos AP
,
Wungjiranirun M
,
Sherman JR
,
Ingram L
,
Batki C
,
Lariviere JP
,
Hamilton JW
.
???displayArticle.abstract???
BACKGROUND: Chronic exposure to excess arsenic in drinking water has been strongly associated with increased risks of multiple cancers, diabetes, heart disease, and reproductive and developmental problems in humans. We previously demonstrated that As, a potent endocrine disruptor at low, environmentally relevant levels, alters steroid signaling at the level of receptor-mediated gene regulation for all five steroid receptors.
OBJECTIVES: The goal of this study was to determine whether As can also disrupt gene regulation via the retinoic acid (RA) receptor (RAR) and/or the thyroid hormone (TH) receptor (TR) and whether these effects are similar to previously observed effects on steroid regulation.
METHODS AND RESULTS: Human embryonic NT2 or rat pituitary GH3 cells were treated with 0.01-5 microM sodium arsenite for 24 hr, with or without RA or TH, respectively, to examine effects of As on receptor-mediated gene transcription. At low, noncytotoxic doses, As significantly altered RAR-dependent gene transcription of a transfected RAR response element-luciferase construct and the native RA-inducible cytochrome P450 CYP26A gene in NT2 cells. Likewise, low-dose As significantly altered expression of a transfected TR response element-luciferase construct and the endogenous TR-regulated type I deiodinase (DIO1) gene in a similar manner in GH3 cells. An amphibian ex vivo tail metamorphosis assay was used to examine whether endocrine disruption by low-dose As could have specific pathophysiologic consequences, because tail metamorphosis is tightly controlled by TH through TR. TH-dependent tail shrinkage was inhibited in a dose-dependent manner by 0.1- 4.0 microM As.
CONCLUSIONS: As had similar effects on RAR- and TR-mediated gene regulation as those previously observed for the steroid receptors, suggesting a common mechanism or action. Arsenic also profoundly affected a TR-dependent developmental process in a model animal system at very low concentrations. Because RAR and TH are critical for both normal human development and adult function and their dysregulation is associated with many disease processes, disruption of these hormone receptor-dependent processes by As is also potentially relevant to human developmental problems and disease risk.
Figure 1. Dose–response curves for As, ATRA, and TH in NT2 and GH3 cells calculated using average values for each dose. Data points represent the mean ± SE of data from three separate experiments. (A) Cytotoxicity of As in NT2 cells exposed to As for 24 hr and assessed by colony-forming assay. Data are expressed as colony formation as a percent of the control. (B) Induction of RARE-luc expression in NT2 cells by ATRA. RARE-luc expression is expressed as mean ± SE luciferase (LU) per gram protein. The median effective concentration (EC50) for ATRA induction is approximately 7 nM. (C) Cytotoxicity of As in GH3 cells assessed essentially as described for (A), except that cells were cultured in media plus stripped serum with or without 10 nM T3. (D) Induction of DIO1 expression by T3 in GH3 cells assessed essentially as described in (C), except that DIO1 mRNA was assessed by RT-PCR. Data are expressed as a percent of the maximum value. See “Methods” for details.
Figure 2. Effects of As on ATRA induction of RARE-luc expression in NT2 cells. Cells were transfected with the RARE-luc construct 24 hr before treatment with 10 nM ATRA with or without simultaneous treatment with As for 24 hr. See “Methods” for details. Data are expressed as mean ± SE of the values from replicates of experiments. Bars that do not have the same letter are significantly different from each other at p < 0.003 using an unpaired t-test.
Figure 3. Effects of As on ATRA induction of CYP26A mRNA expression in NT2 cells. Cells were treated with 10 nM ATRA with or without simultaneous addition of As for 24 hr; mRNA expression was measured by RT-PCR. See “Methods” for details. Data are expressed as mean ± SE of the values from replicates of experiments. Bars that do not have the same letter are significantly different from each other at p < 0.003 using an unpaired t-test.
Figure 4. Effects of As on T3 induction of TRE-luc expression in GH3 cells. Cells were transfected with the TRE-luc construct 24 hr before treatment with 2 nM T3 with or without simultaneous treatment with As for 24 hr. See “Methods” for details. Data are expressed as mean ± SE of the values from replicates of experiments. Bars that do not have the same letter are significantly different from each other at p < 0.003 using an unpaired t-test.
Figure 5. Effects of As on T3 induction of DIO1 mRNA expression in GH3 cells. Cells were treated with 2 nM T3 with or without As, and DIO1 mRNA expression was measured 6 hr (A) or 24 hr (B) after treatment. See “Methods” for details. Data are expressed as mean ± SE of the values from replicates of experiments. Bars that do not have the same letter are significantly different from each other at p < 0.01 using pairwise Student’s t-test analysis.
Figure 6. Effects of T3 on tailfin shrinkage of Xenopus tadpole tails cultured ex vivo as described in “Methods.” Data are expressed as mean ± SE of values from six tails per treatment in three separate experiments.
Figure 7. Effects of As on T3-mediated tail shrinkage in Xenopus tadpole tails cultured ex vivo shown by representive samples from tail resorption experiments. See “Methods” for details. Morphometric software was used to trace the tailfin area (shown in black), which was used to calculate the differences in area for each tail between day 1 and day 4. Results from these experiments are shown in Figures 6 and 8.
Figure 8. Effects of As on T3-mediated tail shrinkage in Xenopus tadpole tails cultured ex vivo. See “Methods” for details. Data are expressed as mean + SE of values from 5–6 individual tails per experiment and 4–8 individual experiments per treatment. Bars that do not have the same letter are significantly different from each other at p < 0.01 using pairwise Student’s t-test analysis.
A,
THE RESULTS OF EXTIRPATION OF THE ANTERIOR LOBE OF THE HYPOPHYSIS AND OF THE THYROID OF RANA PIPIENS LARVAe.
1916, Pubmed
A,
THE RESULTS OF EXTIRPATION OF THE ANTERIOR LOBE OF THE HYPOPHYSIS AND OF THE THYROID OF RANA PIPIENS LARVAe.
1916,
Pubmed
Abernathy,
Arsenic: health effects, mechanisms of actions, and research issues.
1999,
Pubmed
Abernathy,
Health effects and risk assessment of arsenic.
2003,
Pubmed
Abu-Abed,
The retinoic acid-metabolizing enzyme, CYP26A1, is essential for normal hindbrain patterning, vertebral identity, and development of posterior structures.
2001,
Pubmed
Allen,
Oxidative stress by inorganic arsenic: modulation by thyroid hormones in rat.
2003,
Pubmed
Andrew,
Genomic and proteomic profiling of responses to toxic metals in human lung cells.
2003,
Pubmed
Andrew,
Arsenic exposure is associated with decreased DNA repair in vitro and in individuals exposed to drinking water arsenic.
2006,
Pubmed
Aposhian,
Arsenic toxicology: five questions.
2006,
Pubmed
Bastien,
Nuclear retinoid receptors and the transcription of retinoid-target genes.
2004,
Pubmed
Bodwell,
Arsenic disruption of steroid receptor gene activation: Complex dose-response effects are shared by several steroid receptors.
2006,
Pubmed
Bodwell,
Arsenic at very low concentrations alters glucocorticoid receptor (GR)-mediated gene activation but not GR-mediated gene repression: complex dose-response effects are closely correlated with levels of activated GR and require a functional GR DNA binding domain.
2004,
Pubmed
Brown,
The thyroid hormone-induced tail resorption program during Xenopus laevis metamorphosis.
1996,
Pubmed
,
Xenbase
Buchholz,
A dominant-negative thyroid hormone receptor blocks amphibian metamorphosis by retaining corepressors at target genes.
2003,
Pubmed
,
Xenbase
Buchholz,
Molecular and developmental analyses of thyroid hormone receptor function in Xenopus laevis, the African clawed frog.
2006,
Pubmed
,
Xenbase
Buckbinder,
Expression of the Xenopus laevis prolactin and thyrotropin genes during metamorphosis.
1993,
Pubmed
,
Xenbase
Chen,
Chronic inorganic arsenic exposure induces hepatic global and individual gene hypomethylation: implications for arsenic hepatocarcinogenesis.
2004,
Pubmed
Das,
Gene expression changes at metamorphosis induced by thyroid hormone in Xenopus laevis tadpoles.
2006,
Pubmed
,
Xenbase
Davey,
Arsenic as an endocrine disruptor: effects of arsenic on estrogen receptor-mediated gene expression in vivo and in cell culture.
2007,
Pubmed
Du Pasquier,
Developmental cell death during Xenopus metamorphosis involves BID cleavage and caspase 2 and 8 activation.
2006,
Pubmed
,
Xenbase
Galton,
The roles of the iodothyronine deiodinases in mammalian development.
2005,
Pubmed
Göthe,
Mice devoid of all known thyroid hormone receptors are viable but exhibit disorders of the pituitary-thyroid axis, growth, and bone maturation.
1999,
Pubmed
Hermanson,
Nuclear receptor coregulators: multiple modes of modification.
2002,
Pubmed
Hwang,
Induction of cyclin D1 by submicromolar concentrations of arsenite in human epidermal keratinocytes.
2006,
Pubmed
Jakobs,
The promoter of the human type I 5'-deiodinase gene--mapping of the transcription start site and identification of a DR+4 thyroid-hormone-responsive element.
1997,
Pubmed
Kaltreider,
Arsenic alters the function of the glucocorticoid receptor as a transcription factor.
2001,
Pubmed
Karagas,
Incidence of transitional cell carcinoma of the bladder and arsenic exposure in New Hampshire.
2004,
Pubmed
Kitagawa,
Thyroid hormone action: induction of morphological changes and stimulation of cell growth in rat pituitary tumor GH3 cells.
1987,
Pubmed
Kitamura,
Thyroid hormonal activity of the flame retardants tetrabromobisphenol A and tetrachlorobisphenol A.
2002,
Pubmed
Kitchin,
Recent advances in arsenic carcinogenesis: modes of action, animal model systems, and methylated arsenic metabolites.
2001,
Pubmed
Kurokawa,
Regulation of retinoid signalling by receptor polarity and allosteric control of ligand binding.
1994,
Pubmed
Lim,
A thyroid hormone antagonist that inhibits thyroid hormone action in vivo.
2002,
Pubmed
,
Xenbase
Liu,
Global gene expression associated with hepatocarcinogenesis in adult male mice induced by in utero arsenic exposure.
2006,
Pubmed
Liu,
Transplacental arsenic plus postnatal 12-O-teradecanoyl phorbol-13-acetate exposures associated with hepatocarcinogenesis induce similar aberrant gene expression patterns in male and female mouse liver.
2006,
Pubmed
Loudig,
Cytochrome P450RAI(CYP26) promoter: a distinct composite retinoic acid response element underlies the complex regulation of retinoic acid metabolism.
2000,
Pubmed
Mann,
Arsenic trioxide inhibits nuclear receptor function via SEK1/JNK-mediated RXRalpha phosphorylation.
2005,
Pubmed
Mark,
Function of retinoid nuclear receptors: lessons from genetic and pharmacological dissections of the retinoic acid signaling pathway during mouse embryogenesis.
2006,
Pubmed
McKenna,
Minireview: nuclear receptor coactivators--an update.
2002,
Pubmed
Miyazaki,
The effects of gestational arsenic exposure and dietary selenium deficiency on selenium and selenoenzymes in maternal and fetal tissues in mice.
2005,
Pubmed
Mukherjee,
Arsenic contamination in groundwater: a global perspective with emphasis on the Asian scenario.
2006,
Pubmed
Nakajima,
Dual mechanisms governing muscle cell death in tadpole tail during amphibian metamorphosis.
2003,
Pubmed
,
Xenbase
Porterfield,
The role of thyroid hormones in prenatal and neonatal neurological development--current perspectives.
1993,
Pubmed
Raz,
Effects of retinoid and thyroid receptors during development of the inner ear.
1997,
Pubmed
Rossman,
Mechanism of arsenic carcinogenesis: an integrated approach.
2003,
Pubmed
Shaw,
Role of glucocorticoid receptor in acclimation of killifish (Fundulus heteroclitus) to seawater and effects of arsenic.
2007,
Pubmed
Shen,
Fetal onset of aberrant gene expression relevant to pulmonary carcinogenesis in lung adenocarcinoma development induced by in utero arsenic exposure.
2007,
Pubmed
Shenefelt,
Morphogenesis of malformations in hamsters caused by retinoic acid: relation to dose and stage at treatment.
1972,
Pubmed
Shi,
Complex regulation of thyroid hormone action: multiple opportunities for pharmacological intervention.
2002,
Pubmed
Smith,
Increased mortality from lung cancer and bronchiectasis in young adults after exposure to arsenic in utero and in early childhood.
2006,
Pubmed
Smith,
Cancer risks from arsenic in drinking water.
1992,
Pubmed
Stanton,
Arsenic inhibits CFTR-mediated chloride secretion by killifish (Fundulus heteroclitus) opercular membrane.
2006,
Pubmed
THOMPSON,
VITAMIN A AND REPRODUCTION IN RATS.
1964,
Pubmed
Tapio,
Arsenic in the aetiology of cancer.
2006,
Pubmed
Tata,
Requirement for RNA and protein synthesis for induced regression of the tadpole tail in organ culture.
1966,
Pubmed
Tsai,
Molecular mechanisms of action of steroid/thyroid receptor superfamily members.
1994,
Pubmed
Veldhoen,
Detection of environmental endocrine-disruptor effects on gene expression in live Rana catesbeiana tadpoles using a tail fin biopsy technique.
2001,
Pubmed
Vogt,
Effects of arsenite on p53, p21 and cyclin D expression in normal human fibroblasts -- a possible mechanism for arsenite's comutagenicity.
2001,
Pubmed
Waalkes,
Transplacental carcinogenicity of inorganic arsenic in the drinking water: induction of hepatic, ovarian, pulmonary, and adrenal tumors in mice.
2003,
Pubmed
Waalkes,
Induction of tumors of the liver, lung, ovary and adrenal in adult mice after brief maternal gestational exposure to inorganic arsenic: promotional effects of postnatal phorbol ester exposure on hepatic and pulmonary, but not dermal cancers.
2004,
Pubmed
Waalkes,
Estrogen signaling in livers of male mice with hepatocellular carcinoma induced by exposure to arsenic in utero.
2004,
Pubmed
Wasserman,
Water arsenic exposure and children's intellectual function in Araihazar, Bangladesh.
2004,
Pubmed
Watanabe,
Effects of arsenic on younger generations.
2003,
Pubmed
White,
Identification of the retinoic acid-inducible all-trans-retinoic acid 4-hydroxylase.
1996,
Pubmed
White,
cDNA cloning of human retinoic acid-metabolizing enzyme (hP450RAI) identifies a novel family of cytochromes P450.
1997,
Pubmed
Wolbach,
Nutrition Classics. The Journal of Experimental Medicine 42: 753-77, 1925. Tissue changes following deprivation of fat-soluble A vitamin. S. Burt Wolbach and Percy R. Howe.
1978,
Pubmed
Wong,
Coordinated regulation of and transcriptional activation by Xenopus thyroid hormone and retinoid X receptors.
1995,
Pubmed
,
Xenbase
Yamamoto,
Cancer induction by an organic arsenic compound, dimethylarsinic acid (cacodylic acid), in F344/DuCrj rats after pretreatment with five carcinogens.
1995,
Pubmed
Yaoita,
A correlation of thyroid hormone receptor gene expression with amphibian metamorphosis.
1990,
Pubmed
,
Xenbase
Zhang,
The mechanism of action of thyroid hormones.
2000,
Pubmed