METHODS: Patients with CCAC of the uterine cervix or vagina born after 1946 and diagnosed in the period 1969-2005, were identified through the Nationwide network and registry of histo- and cytopathology in the Netherlands and from 2003 onwards through the Netherlands Cancer Registry. Exposure data and clinical data were obtained by means of questionnaires and medical records. The histology slides of tumours were reviewed. For the patients who did not provide consent, only the date of diagnosis and age at diagnosis were known (n = 10).
RESULTS: Up to 2005, 144 CCAC patients were registered. Age at diagnosis varied from 8-54 years (mean: 28 years). In the years 1981-2000, the number of new diagnoses in 4 successive 5-year periods was fairly stable (n=26-30) but it was considerably lower in 2001-2005 (n=13). Of the patients whose history of intrauterine exposure to DES was known, 62% had been exposed (76/122). The mean age at diagnosis was 24 years for exposed patients compared to 32 years for non-exposed patients. The 10-year survival rate was 78% (95% CI: 68-87) for exposed and 69% (95% CI: 56-82) for non-exposed patients.
CONCLUSION: Since 2000, the incidence of CCAC of the vagina and cervix has decreased markedly compared to the situation in the 1980s and 1990 s. In particular, the number of patients with CCAC exposed in utero to DES has decreased. Whether this decrease shall continue over the coming years remains to be seen.
Sources and more information
Vaginal and cervical cancer due to diethylstilbestrol (DES); end epidemic, Ned Tijdschr Geneeskd. 2009;153:A366. NCBI PMID: 19857300.
Environmental Estrogens and Obesity, Molecular Cellular Endocrinology, 2009
2009 Study Abstract
Diethylstilbestrol DES, a potent synthetic estrogen, was widely prescribed to pregnant women from the 1940s through the 1970s with the mistaken belief that it could prevent threatened miscarriages. It was estimated that a range of 2 to 8 million pregnancies worldwide were exposed to DES. Today, it is well known that prenatal DES treatment resulted in a low but significant increase in neoplastic lesions, and a high incidence of benign lesions in both the male and female offspring exposed during fetal life. To study the mechanisms involved in DES toxicity, we developed experimental mouse models of perinatal (prenatal or neonatal) DES exposure over 30 years ago . Outbred CD-1 mice were treated with DES by subcutaneous injections on days 9–16 of gestation (the period of major organogenesis in the mouse) or days 1–5 of neonatal life (a period of cellular differentiation of the reproductive tract, and a critical period of immune, behavioral, and adipocyte differentiation). These perinatal DES animal models have successfully duplicated, and in some cases, predicted, many of the alterations (structural, function, cellular and molecular) observed in similarly DES- exposed humans.
Although the data summarized in this review describes only neonatal exposure to a high dose of DES, lower doses and exposure during prenatal life have also been shown to be associated with obesity later in life. Interestingly, high prenatal DES doses caused lower birth weight compared to controls, followed by a “catch-up period”, and finally resulted in obesity; low prenatal DES doses had no effect on birth weight but it still resulted in obesity later in life . Thus, it appears that the effects of DES on adipocytes may depend on the time of exposure and the dose, and that multiple mechanisms maybe altered resulting in the same obesity phenotype.
Environmental Estrogens and Obesity, NCBI PMCID: PMC2682588, Retha R. Newbold,1 Elizabeth Padilla-Banks, and Wendy N. Jefferson, Mol Cell Endocrinol. Author manuscript; available in PMC 2010 May 25.
DES alters the expression levels of Dnmts and DNA methylation
2009 Study Abstract
Perinatal exposure to diethylstilbestrol (DES) can have numerous adverse effects on the reproductive organs later in life, such as vaginal clear-cell adenocarcinoma. Epigenetic processes including DNA methylation may be involved in the mechanisms.
We subcutaneously injected DES to neonatal C57BL/6 mice. At days 5, 14, and 30, expressions of DNA methyltransferases (Dnmts) Dnmt1, Dnmt3a, and Dnmt3b, and transcription factors Sp1 and Sp3 were examined. We also performed restriction landmark genomic scanning (RLGS) to detect aberrant DNA methylation. Real-time RT-PCR revealed that expressions of Dnmt1, Dnmt3b, and Sp3 were decreased at day 5 in DES-treated mice, and that those of Dnmt1, Dnmt3a, and Sp1 were also decreased at day 14. RLGS analysis revealed that 5 genomic loci were demethylated, and 5 other loci were methylated by DES treatment. Two loci were cloned, and differential DNA methylation was quantified.
Our results indicated that DES altered the expression levels of Dnmts and DNA methylation.
Neonatal exposure to diethylstilbestrol alters expression of DNA methyltransferases and methylation of genomic DNA in the mouse uterus, Sato K1, Fukata H, Kogo Y, Ohgane J, Shiota K, Mori C. Endocr J. 2009;56(1):131-9. Epub 2008 Nov 8. NCBI PMID: 18997445.
Nr0b2 reduces testosterone levels after Diethylstilbestrol exposure
2009 Study Summary
Studies in rodents have shown that male sexual function can be disrupted by fetal or neonatal administration of compounds that alter endocrine homeostasis, such as the synthetic nonsteroidal estrogen Diethylstilbestrol (DES). Although the molecular basis for this effect remains unknown, estrogen receptors likely play a critical role in mediating DES-induced infertility. Recently, we showed that the orphan nuclear receptor small heterodimer partner (Nr0b2), which is both a target gene and a transcriptional repressor of estrogen receptors, controls testicular function by regulating germ cell entry into meiosis and testosterone synthesis. We therefore hypothesized that some of the harmful effects of DES on testes could be mediated through Nr0b2. Here, we present data demonstrating that Nr0b2 deficiency protected mice against the negative effects of DES on testis development and function. During postnatal development, Nr0b2-null mice were resistant to DES-mediated inhibition of germ cell differentiation, which may be the result of interference by Nr0b2 with retinoid signals that control meiosis. Adult Nr0b2-null male mice were also protected against the effects of DES; however, we suggest that this phenomenon was due to the removal of the repressive effects of Nr0b2 on steroidogenesis. Together, these data demonstrate that Nr0b2 plays a critical role in the pathophysiological changes induced by DES in the mouse testis.
Our results demonstrated that Nr0b2 plays a major role in the DES signaling pathway, which affects the development and function of the male reproductive system. We showed that Nr0b2L male mice were protected against the deleterious effects of DES, as they were still able to reproduce even when exposed to high doses of DES. This is caused by the multiple actions of Nr0b2 during testicular development. First, in neonatal animals, Nr0b2 controls germ cell differentiation through inhibition of the retinoid pathway. Nr0b2 regulates the expression of genes involved in the entry and progression of meiosis, such as Stra8 and Nanos3. It also affects meiosis through regulation of the expression of the histone methyltransferase G9a and the subsequent modification of H3K9 methylation marks. These alterations in methylation upon DES exposure induce abnormal chromosomal complexes favoring germ cell apoptosis and could affect the meiosis process. Next to these effects, which appear to be mediated through the estrogenic pathway, DES seems to inhibit germ cell differentiation at P10 through estrogen-independent pathways, as shown by Oct3/4 deregulation specifically in DES-treated mice. Second, in adult animals, the effect of Nr0b2 was dependent on the inhibition of testosterone production, leading to germ cell death. Together, our present data define Nr0b2 as one of the major actors in the molecular events leading to DES-mediated male infertility.
Sources and Full Study
The orphan nuclear receptor small heterodimer partner mediates male infertility induced by diethylstilbestrol in mice, NCBI, PMID: 19884658, 2009 Dec.
Full study The Journal of Clinical Investigation. 2009;119(12):3752-3764. doi:10.1172/JCI38521. PMC2786790. 2009 Dec.
Tall Girls, Short Boys, and the Medical Industry’s Quest to Manipulate Height
“Normal at Any Cost” is the story, told decade by decade, of medical attempts to tinker with one inherited characteristic: height. It reveals the way drug companies redefined normal in order to expand markets, and how the best motives and worst motives combined to result in widespread experimentation on children. We think the temptations to tamper with heredity are just beginning.
Susan Cohen‘s book tells the horrible story of drug use to adjust the height of adolescent boys and girls who were threatening to be short, or tall, adults. DES was prescribed to prevent girls from growing “too tall.”
Sources and book reviews
At What Height, Happiness? A Medical Tale, DES Action.
Normal at Any Cost: Tall Girls, Short Boys, and the Medical Industry’s Quest to Manipulate Height, N Engl J Med 2009.
Juin 2009, Côté-mômes rencontrait Abert Algoud, journaliste, écrivain, scénariste, chroniqueur de radio et de télé et humoriste. Très concerné, Abert Algoud s’implique depuis plus de 20 ans dans le combat des autistes pour mener leurs vies le plus dignement possible.
Prenatal exposure to bisphenol a at environmentally relevant doses adversely affects the murine female reproductive tract later in life
Background: Exposure to endocrine-disrupting chemicals during critical developmental periods causes adverse consequences later in life; an example is prenatal exposure to the pharmaceutical diethylstilbestrol (DES). Bisphenol-A (BPA), an environmental estrogen used in the synthesis of plastics, is of concern because its chemical structure resembles that of DES, and it is a “high-volume production” chemical with widespread human exposure.
Objective: In this study we investigated whether prenatal Bisphenol-A causes long-term adverse effects in female reproductive tissues in an experimental animal model previously shown useful in studying effects of prenatal DES.
Methods: Timed pregnant CD-1 mice were treated on days 9-16 of gestation with BPA (0.1, 1, 10, 100, or 1,000 mug/kg/day). After delivery, pups were held for 18 months; reproductive tissues were then evaluated.
Results: Ovarian cysts were significantly increased in the 1-mug/kg BPA group; ovarian cyst-adenomas were seen in the other three BPA-treated groups but not in corn-oil controls. We observed increased progressive proliferative lesions of the oviduct after BPA treatment, similar to those described in response to DES. Further, although not statistically different from the controls, prominent mesonephric (Wolffian) remnants and squamous metaplasia of the uterus, as well as vaginal adenosis, were present in BPA-treated mice, similar to lesions reported following DES treatment. More severe pathologies observed in some BPA-treated animals included atypical hyperplasia and stromal polyps of the uterus; sarcoma of the uterine cervix; and mammary adenocarcinoma. We did not observe these lesions in controls.
Conclusion: These data suggest that BPA causes long-term adverse reproductive and carcinogenic effects if exposure occurs during critical periods of differentiation.
Prenatal exposure to bisphenol a at environmentally relevant doses adversely affects the murine female reproductive tract later in life., Environ Health Perspect ; 117(6):879-85. doi: 10.1289/ehp.0800045, 19590677, 2009 Jun.
How diethylstilbestrol DES and other endocrine disrupting chemical EDCs affect murine female reproductive tract development and cancer at the molecular level
Growing concerns over endocrine disrupting chemicals (EDCs) and their effects on human fetal development and adult health have promoted research into the underlying molecular mechanisms of endocrine disruption. Gene targeting technology has allowed insight into the genetic pathways governing reproductive tract development and how exposure to EDCs during a critical developmental window can alter reproductive tract development, potentially forming the basis for adult diseases. This review primarily uses Diethylstilbestrol (DES) as a model agent for EDCs and discusses the recent progress elucidating how DES and other EDCs affect murine female reproductive tract development and cancer at the molecular level.
Endocrine disruptors in female reproductive tract development and carcinogenesis, NCBI, PMID: 19709900, 2009 Sep;20(7):357-63. doi: 10.1016/j.tem.2009.03.009. Epub 2009 Aug 25.
Full text – NCBI, PMC2774851 Trends Endocrinol Metab. Sep 2009; 20(7): 357–363.
DES has a dual mechanism of action as an endocrine disruptor
Diethylstilbestrol (DES) is a nonsteroidal estrogen that induces developmental anomalies of the female reproductive tract. The homeobox gene HOXA10 controls uterine organogenesis, and its expression is altered after in utero DES exposure. We hypothesized that an epigenetic mechanism underlies DES-mediated alterations in HOXA10 expression. We analyzed the expression pattern and methylation profile of HOXA10 after DES exposure. Expression of HOXA10 is increased in human endometrial cells after DES exposure, whereas Hoxa10 expression is repressed and shifted caudally from its normal location in mice exposed in utero. Cytosine guanine dinucleotide methylation frequency in the Hoxa10 intron was higher in DES-exposed offspring compared with controls (P = 0.017). The methylation level of Hoxa10 was also higher in the caudal portion of the uterus after DES exposure at the promoter and intron (P < 0.01). These changes were accompanied by increased expression of DNA methyltransferases 1 and 3b. No changes in methylation were observed after in vitro or adult DES exposure. DES has a dual mechanism of action as an endocrine disruptor; DES functions as a classical estrogen and directly stimulates HOXA10 expression with short-term exposure, however, in utero exposure results in hypermethylation of the HOXA10 gene and long-term altered HOXA10 expression. We identify hypermethylation as a novel mechanism of DES-induced altered developmental programming.
Hypermethylation of homeobox A10 by in utero diethylstilbestrol exposure: an epigenetic mechanism for altered developmental programming, NCBI, PMID: 19299448, 2009 Jul;150(7):3376-82. doi: 10.1210/en.2009-0071. Epub 2009 Mar 19
Full text – The Endocrine Society, Endocrinology, 10.1210/en.2009-0071, Volume 150 Issue 7 | July 1, 2009.
Women exposed to DES in utero during critical periods of reproductive tract development developed several types of reproductive tract abnormalities, as well as an increased incidence of cervical-vaginal cancer later in life
2009 Study Abstract
Ovarian Follicular Development and the Environment
The ovarian follicle is the functional unit of the ovary and is comprised of an oocyte surrounded and supported by the somatic granulosa and theca cells (28). The health of the follicle can impact the health of the woman as well as the health of her offspring. For example, decreased numbers of follicles, multiple eggs per follicle and incomplete follicular development can all result in decreased fertility. The precise mechanisms involved in early ovarian follicle formation are not known, but are essential in organizing the fetal ovary and establishing the postnatal follicle number that will provide the female with sufficient oocytes for a lifetime of fertility.
Estrogen and activin are two known factors that play an important role in regulating oocyte and follicle development and function and aberrant development and ovarian pathologies are observed in mice exposed to neonatal estrogen or activin. Neonatal exposure of rats to estradiol benzoate has been show to delay follicle and interstitial development. Neonatal exposure to diethylstilbestrol (DES) or the natural estrogen estradiol (E2) results in lack of corpora lutea in adult mice, suggesting that these effects persist beyond reproductive tract development and impact fertility in the adult. Neonatal exposure to DES, E2, or the phytoestrogen genistein also induces formation of multi-oocytic follicles in mice – an effect that is also reported in alligators exposed to environmental estrogenic contaminants. Additionally, activin administered during the critical, postnatal period of primordial follicle formation changes the number of postnatal follicles. Current mechanistic studies are exploring whether neonatal estrogen exposure alters activin signaling in the ovary; preliminary findings of decreased activin subunit gene expression and impacted activin signaling in the mouse ovary support this hypothesis.
Studies have now been extended beyond DES to demonstrate that other environmental estrogens reprogram gene expression in the uterus : exposure to genistein and BPA during the period of maximum sensitivity to developmental programming induces the expression of the estrogen-responsive genes calbindin and progesterone receptor. Neonatal BPA exposure attenuated estrogen-responsive genes whereas genistein exposure induced an even higher level of estrogen responsiveness than DES exposure. In contrast to DES, exposure to these environmental estrogens does not disrupt ovarian function in adult females, which continue to cycle normally.
PMC, Proceedings of the Summit on Environmental Challenges to Reproductive Health and Fertility: executive summary, PMC2440710, Feb 1, 2009.