Estrogen In Tissues Research Articles

Genetic variant in the CYP19 gene and recurrent spontaneous abortions

BackgroundThe CYP19 gene encodes aromatase, a key cytochrome P450 enzyme that converts androgens to estrogens in the ovarian tissues of premenopausal women. A polymorphic variation C/T in the CYP19 gene, rs10046, results in modified estrogen levels through increasing aromatase activity. In the present study, we investigated the possible association between the rs10046 polymorphism of the CYP19 gene and the risk of recurrent spontaneous abortions (RSA) in the Caucasian Greek population. MethodsIn the prospective case-control study 120 patients and 100 healthy controls were studied. Women who had at least three unexplained spontaneous abortions before 20weeks of gestation were included in the patient group, while the control group consisted of women with at least two successful pregnancies and without history of abortions. The PCR-RFLP method was used in order to genotype the subjects. ResultsThe frequencies of CC, CT, TT genotypes of the rs10046 variant were 0.275, 0.500, 0.225, respectively, in the patient group and 0.210, 0.480, 0.310, respectively, in the control group. The allele frequencies were 0.525 and 0.475 for C and T, respectively, in the patient group and 0.450 and 0.550 for C and T, respectively, in the control group. Statistical analysis of these results indicated that there are no significant differences in genotype (P=0.2897) or in allele frequencies [P=0.1261, OR (95%)=1.351(0.9269–1.969)] between RSA patient and the control groups. ConclusionThe rs10046 polymorphism of CYP19 gene seems that is not an important contributing factor in the aetiology of RSA in the Greek Caucasian population.

The presence and impact of estrogen metabolism on the biology of triple-negative breast cancer.

While triple-negative breast cancer (TNBC) is negative for estrogen receptor alpha, a substantial proportion of carcinomas express estrogen receptor beta (ERβ); consequently, estrogen actions and metabolism may be relevant in this cancer subtype. A cohort of 81 TNBC patients from Tohoku University Hospital, Japan were characterised with regard to the expression of estrogen receptor beta and enzymes known to modulate levels of estrogens in breast and other tissues (Aromatase, 17-beta- Hydroxysteroid dehydrogenases 1, 2 and 6). This was done at the protein level by means of immunohistochemistry. As this cohort has been previously characterised for androgens, this also allows for comparison between the expressions of estrogen-related proteins and of androgen-related proteins. Preliminary mechanistic studies in cell culture were also undertaken. 17βHSD2 was detected in the highest number of cases followed by 17βHSD1, 17βHSD6 and aromatase. When comparing the expression of ERβ with that of the enzymes, it was positively correlated with the expression of 17βHSD6 (p<0.05) and trended towards correlation with dual expression of 17βHSD1 and 2 (p<0.07). 17βHSD1 was associated with significantly reduced tumour volume (p=0.0025), while ERβ was associated with a trend towards reduced lymphovascular invasion, (p<0.061). Interestingly, in survival analysis, 17βHSD6 expression was the only one of these five factors that influenced survival, with positive samples being associated with longer disease-free survival compared to those that were negative for 17βHSD6 (p<0.05). In assessing associations with expression of proteins in the androgenic pathway, expression of aromatase appeared to be associated with androgenic pathways in TNBC patients (p<0.05). Due to this association and the potential relevance to androgen-directed therapies in TNBC, we evaluated this interaction in vitro. We observed androgen-dependent upregulation of aromatase and ERβ in a subset of AR expressing TNBC cell lines (MDA-MB-453, SUM-185-PE and MFM-223). Overall this study suggests the presence of, and a potential protective effect of estrogens in TNBC.

Effects of Chronic Estrogen Administration in the Ventromedial Nucleus of the Hypothalamus (VMH) on Fat and Bone Metabolism in Ovariectomized Rats.

Estrogen deficiency after ovariectomy (OVX) results in increased adiposity and bone loss, which can be prevented by systemic 17-β estradiol (E2) replacement. Studies in transgenic mice suggested that in addition to direct actions of estrogen in peripheral tissues, also estrogen signaling in the hypothalamus regulates fat distribution and bone metabolism. We hypothesized that the protective effect of systemic E2 on fat and bone metabolism in the OVX model is partly mediated through the ventromedial nucleus of the hypothalamus (VMH). To test this hypothesis, we determined the effect of systemic, central, and targeted VMH administration of E2 on fat and bone metabolism in OVX rats. Subcutaneous administration of E2 for 4 weeks decreased body weight, gonadal and perirenal fat, and bone formation rate in OVX rats. This effect was completely mimicked by intracerebroventricular injections of E2, once every 4 days for 4 weeks. Administration of E2 locally in the VMH by retromicrodialysis (3 h) acutely increased expression of the lipolytic gene hormone-sensitive lipase in gonadal and perirenal fat. Finally, chronic administration of E2 in the VMH for 8 weeks decreased perirenal fat but did not affect body weight, trabecular bone volume, or cortical thickness. In conclusion, we demonstrated that intracerebroventricular E2 replacement reduces body weight gain, ameliorates intraabdominal fat accumulation, and reduces bone formation in the OVX rats. E2 administration selectively in the VMH also reduced intraabdominal fat but did not affect bone metabolism.

Synchronous Multiple Lung Adenocarcinomas: Estrogen Concentration in Peripheral Lung.

BackgroundThe detection rate of synchronous multiple lung adenocarcinomas (SMLA), which display multiple ground glass opacity nodules in the peripheral lung, is increasing due to advances in high resolution computed tomography. The backgrounds of multicentric development of adenocarcinoma are unknown. In this study, we quantitated estrogen concentration in the peripheral lungs of postmenopausal female patients with SMLA.MethodsThe tissue concentration of estrogens (estrone [E1] and estdadiol [E2]) in the noncancerous peripheral lung were measured with liquid chromatography/electrospray tandem mass spectrometry in postmenopausal female patients with lung adenocarcinoma. The expression levels of CYP19A1 in the normal lung were also quantitated with real-time PCR. Thirty patients with SMLA and 79 cases of control patients with single lung adenocarcinoma were analyzed.ResultsThe concentrations of E1 and E2 in the noncancerous tissue were significantly higher in SMLA cases than control cases (P = 0.004 and P = 0.02, respectively). The minor allele (A) of single nucleotide polymorphism rs3764221 were significantly associated with higher concentration of E1 and E2 (P = 0.002 and P = 0.01, respectively) and higher CYP19A1 mRNA expression (P = 0.03).ConclusionThe tissue estrogen concentration of peripheral lung was significantly higher in SMLA than control cases. The high concentration of estrogen may be one of the causes of multicentric development of peripheral lung adenocarcinomas.

A high dose mode of action for tetrabromobisphenol A-induced uterine adenocarcinomas in Wistar Han rats: A critical evaluation of key events in an adverse outcome pathway framework

TBBPA is a non-genotoxic flame retardant used to improve fire safety in a wide variety of consumer products. Estimated human exposures to TBBPA are very low (<0.000084 mg/kg-day), relative to the doses (500 and 1000 mg/kg-day of TBBPA) administered in a recent bioassay that resulted in uterine tumors in Wistar Han rats following chronic exposure. As part of an effort to characterize the relevance of the uterine tumors to humans, data and biological knowledge relevant to the progression of events associated with TBBPA-induced uterine tumors in female rats were organized in an adverse outcome pathway (AOP) framework. Based on a review of possible MOAs for chemically induced uterine tumors and available TBBPA data sets, a plausible molecular initiating event (MIE) was the ability of TBBPA to bind to and inhibit estrogen sulfotransferases, the enzymes responsible for sulfation of estradiol. Subsequent key events in the AOP, including increased bioavailability of unconjugated estrogens in uterine tissue, would occur as a result of decreased sulfation, leading to a disruption in estrogen homeostasis, increased expression of estrogen responsive genes, cell proliferation, and hyperplasia. Available data support subsequent key events, including generation of reactive quinones from the metabolism of estrogens, followed by DNA damage that could contribute to the development of uterine tumors. Uncertainties associated with human relevance are highlighted by potential strain/species sensitivities to development of uterine tumors, as well as the characterization of a dose-dependent MIE. For the latter, it was determined that the TBBPA metabolic profile is altered at high doses (such as those used in the cancer bioassay), and thus an MIE that is only operative under repeated high dose, administration. The MIE and subsequent key events for the development of TBBPA-induced uterine tumors are not feasible in humans given differences in the kinetic and dynamic factors associated with high dose exposures in rats relative to human exposure levels to TBBPA.

Modulation of Aromatase by Phytoestrogens.

The aromatase enzyme catalyzes the conversion of androgens to estrogens in many human tissues. Estrogens are known to stimulate cellular proliferation associated with certain cancers and protect against adverse symptoms during the peri- and postmenopausal intervals. Phytoestrogens are a group of plant derived naturally occurring compounds that have chemical structures similar to estrogen. Since phytoestrogens are known to be constituents of animal/human food sources, these compounds have received increased research attention. Phytoestrogens may contribute to decreased cancer risk by the inhibition of aromatase enzyme activity and CYP19 gene expression in human tissues. This review covers (a) the aromatase enzyme (historical descriptions on function, activity, and gene characteristics), (b) phytoestrogens in their classifications and applications to human health, and (c) a chronological coverage of aromatase activity modulated by phytoestrogens from the early 1980s to 2015. In general, phytoestrogens act as aromatase inhibitors by (a) decreasing aromatase gene expression, (b) inhibiting the aromatase enzyme itself, or (c) in some cases acting at both levels of regulation. The findings presented herein are consistent with estrogen's impact on health and phytoestrogen's potential as anticancer treatments, but well-controlled, large-scale studies are warranted to determine the effectiveness of phytoestrogens on breast cancer and age-related diseases.

Consumption of the Placenta in the Postpartum Period

Postpartum women are consuming their placentas to achieve claimed health benefits, including improved mood, energy, and lactation. Strong scientific evidence to substantiate these claims is lacking. Self-reported benefits from some women include improved mood and lactation; animal models suggest there may be an analgesic effect. Possible risks include infection, thromboembolism from estrogens in placental tissue, and accumulation of environmental toxins. Women’s health care providers should be aware of this practice to help women make informed decisions.

Exercise induces the synthesis of estrogen in ovariectomized Sprague–Dawley rats ventricular myocardium trough increase expression of CYP19aromatase

Postmenopausal women have complex health problems related to hormonal status. Recently there are controversial issues about hormonal therapy for heart diseases. Another alternative to overcome heart disease in postmenopause is exercise. Several studies have reported that exercise can induce estrogen synthesis in the extragonadal organs. The aim of this study is to know the estrogen synthesis process by aromatization in the ventricular myocardium of ovariectomized rats after exercise. The research design of this study was experimental with post-test only control group design for the tissue levels of estrogen and testosterone as well as CYP19aromatase and COX2 expression. The subjects of this study were 68 healthy female Sprague-Dawley rats aged 4–5 weeks given exercise using a rat treadmill for 12 weeks, 5×/week by graded exercise at 17–23 m/min for 10–60 min with 5–10% inclination. Tissue estrogen and testosterone levels were determined using ECLIA, and to determine CYP19aromatase and COX2 expression using IHC method. The ovariectomized and exercised groups lead to higher tissue estrogen levels (36.67 ± 9.23 compared to 24.56 ± 3.33 pg/dL), higher CYP19aromatase expression (44.43 ± 5.56 %) compared to group without exercise (34.09 ± 2.62 %), as well as COX2 47.34 ± 1.86 % compared to 32.02 + 2.81 %. No significant difference was found in tissue testosterone levels. We analyze the data with multiple regressions to determine the influence of these three variables. Tissue estrogen levels were predictable from CYP19aromatase and COX2 expression but not tissue testosterone levels. Multiple regressions r square showed that 51.2 % of the total variation in estrogen tissue levels could be accounted for by CYP19aromatase and COX2 expression. The present study found that 12-week exercise with treadmill prevents ovariectomized-induced deterioration of ventricular myocardium estrogen levels of rats. It is likely that the beneficial effects of exercise were exerted through the increase of expression of CYP19aromatase and COX2 in ventricular myocardium ovariectomized rats. Nevertheless, the precise mechanisms by which exercise prevents deterioration of estrogen levels in ventricular myocardium, which have consequences in the prevention of heart disease, require further investigations. Exercise of postovariectomized rat leads to the ventricular myocardium estrogen synthesis through the increase of CYP19aromatase and COX2 expression.

ATPS-13AROMATASE EXPRESSION IN HIGH GRADE GLIOMAS: A POTENTIAL NEW TARGET FOR THERAPY

The prognosis for patients with malignant gliomas remains dismal and novel therapeutic agents are urgently needed. We investigated the potential role of aromatase (CYP19A1), an enzyme that catalyzes the conversion of androgens to estrogens in peripheral tissues, as a target for the treatment of CNS malignancies, as well as brain disposition and anti-tumor efficacy of letrozole, an aromatase inhibitor. Two recent discoveries from our lab have immediate clinical import. First, we analyzed the expression of aromatase in 67 human biopsy samples obtained from the University of Cincinnati Cancer Institute Tumor bank. De-identified frozen tissue samples (containing at least 50 % tumor cells, confirmed by HE i.p. injections) had long term suppression with overall survival exceeding 65 days and no signs of overt toxicity. In contrast, control untreated rats (N = 6, 2ml/kg vehicle i.p. injections) developed significant morbidity/mortality in 15-20 days. Overall, our studies strongly suggest that aromatase is a new “druggable target” for treatment of HGGs and that letrozole can potentially be readily used for this purpose.

Recent developments in steroidal and nonsteroidal aromatase inhibitors for the chemoprevention of estrogen-dependent breast cancer

Aromatase, a cytochrome P450 enzyme complex present in breast tissues, plays a significant role in the biosynthesis of important endogenous estrogens from androgens. The source of estrogen production in breast cancer tissues is intra-tumoral aromatase, and inhibition of aromatase may inhibit the growth stimulation effect of estrogens in breast cancer tissues. Consequently, aromatase is considered a useful therapeutic target in the treatment and prevention of estrogen-dependent breast cancer. Recently, different natural products and synthetic compounds have been rapidly developed, studied, and evaluated for aromatase inhibitory activity. Aromatase inhibitors are classified into two categories on the basis of their chemical structures, i.e., steroidal and nonsteroidal aromatase inhibitors. This review highlights the synthetic steroidal and nonsteroidal aromatase inhibitors reported in the literature in the last few years and will aid medicinal chemists in the design and synthesis of novel and pharmacologically-potent aromatase inhibitors for the treatment of breast cancer.

Abstract 5038: Role of the short isoform of the progesterone receptor in hormone-regulated invasiveness of breast cancer cells at pre- and post-menopausal levels of estrogen and progesterone

Abstract Introduction: High expression of the progesterone receptor (PR) isoform A, relative to PR isoform B, is clinically associated with progression to invasiveness in estrogen receptor (ER)-positive breast cancer and lower disease free survival. This is apparently at odds with in vitro studies implicating PR-B in mediating progestin-induced invasiveness. As the in vitro studies were conducted in the absence of estrogen (E2) signaling, we considered the possibility that modulation of E2 action may comprise a distinct aspect of the regulation of invasiveness by progestins. Methods: Standard ER+/PR+ and ER+/PR+/HER2+ breast cancer cell line models were studied, as well as recombinant cells expressing a single PR isoform. For the progestin hyper-sensitization studies, PR-A was over-expressed by lentiviral infection. Hormonal influence on invasiveness was studied using the Boyden chamber assay. Gene expression was examined by Illumina DNA microarray analysis and by Taq-Man real time RT-PCR assays. Results: E2 strongly suppressed invasiveness of ER+ breast cancer cells at concentrations below 0.01nM. At low (&amp;lt; 1 nM) concentrations, progesterone, R5020 and medroxyprogesterone acetate completely abrogated the inhibition of invasiveness by E2. It was only in a higher (5nM - 50 nM) concentration range that progestins induced invasiveness in the absence of E2. The ability of progestins to rescue invasiveness from E2 regulation was exclusively mediated by PR-A whereas PR-B mediated the E2-independent component of progestin-induced invasiveness. Overexpression of PR-A in PR-A+/PR-B+ cells lowered the progestin concentration needed to completely rescue invasiveness (to &amp;lt; 0.2 nM). Progesterone opposed E2 regulation of a distinctive set of genes, exclusively through PR-A. Conclusion: At relatively low concentrations, progesterone influences breast cancer cell invasiveness by rescuing it from E2 regulation via PR-A, whereas at higher concentrations the hormone also induces invasiveness independent of E2 signaling, through PR-B. The findings establish a direct functional link between PR-A and progression of luminal breast cancer in the context of the entire range of pre- and post-menopausal plasma and breast tissue estrogen and progesterone levels. Citation Format: Thomas B. McFall, Mugdha Patki, Rayna Rosati, Manohar Ratnam. Role of the short isoform of the progesterone receptor in hormone-regulated invasiveness of breast cancer cells at pre- and post-menopausal levels of estrogen and progesterone. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5038. doi:10.1158/1538-7445.AM2015-5038

Serum estrone concentration, estrone sulfate/estrone ratio and BMI are associated with human epidermal growth factor receptor 2 and progesterone receptor status in postmenopausal primary breast cancer patients suffering invasive ductal carcinoma.

BackgroundWe investigated in postmenopausal women with primary breast cancer prior to surgical intervention whether, serum levels of different steroid hormones and hormonal precursors associated with tumor tissue estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) status.MethodsWe enrolled 1,042 patients suffering invasive ductal carcinoma undergoing surgical resection in the National Institute of Oncology, Hungary between 2003 and 2011. Serum parameters were measured by RIA/IRMA assays; tumor tissue ER, PR and HER2 status was assessed histologically. Patients were classified according to tumor receptor status. Case–case analysis subjects were categorized into four subgroups based on serum hormone concentrations in ER, PR and HER2 receptor-negative cases, respectively.ResultsSerum estrone sulfate and dehydroepiandrosterone sulfate levels correlated with each other and also with serum estrone and estradiol levels. According to case–case study the odds ratios in the highest quartile were 1.517 (p = 0.0305, Ptrend = 0.0394) for androstenedione, 1.495 (p = 0.0317, Ptrend < 0.0105) for estrone and 0.654 (p = 0.0273, Ptrend < 0.0151) for estrone sulfate/estrone ratio in PR+ vs. PR− tumors. Regarding HER2 status (HER2+ vs. HER2−), the odds ratios for estrone, estrone sulfate and estrone sulfate/estrone ratio were 0.530 (p = 0.0234, Ptrend = 0.0595), 2.438 (p = 0.0042, Ptrend < 0.0066) and 3.118 (p = 0.0001, Ptrend < 0.0001) in the highest quartile, respectively. Of note significantly increased BMI associates with PR+ and ER +/PR+ status while significantly decreased BMI was observed in HER2+ cases.ConclusionsTaken together, measurement of serum estrone and estrone sulfate concentrations prior to surgical intervention might support the individualization of regime in postmenopausal primary breast cancer patients.

Abstract P1-12-11: Prospective study of aromatase inhibitor-induced bone loss and lipid levels in early hormone sensitive breast cancer treated with AI during 8 years

Abstract Introduction: AI’s are the preferred adjuvant hormonal treatment for postmenopausal(PM) estrogen receptor positive (ER+) disease because of the improved efficacy over tamoxifen in terms of disease free survival and reduction in distant recurrence. AI’s have been associated with increased bone loss by blocking peripheral tissue estrogen synthesis, arthralgia and a negative effect on lipid metabolism. In this prospective study we wanted to assess the amount of bone loss and the changes in lipid levels in early breast cancer(EBC) patients (pts) treated with 3yrs of extended letrozole as part of the SOLE study after 5 yrs of adjuvant AI. Objectives: the primary objective of the study is to calculate the mean percentage change in bone mineral density(BMD)(g/cm2) after 8 yrs of letrozole and to compare between the continuous(C) and intermittent(I) intake of letrozole. The secondary objectives are to correlate the mean change in BMD with the initial value, BMI and to describe the evolution of the lipid levels in both groups. Patients and methods: BMD was measured at the lumbar spine (L2-4) and hip by dual energy X-ray absorptiometry (DEXA) in PM pts. with ER+ EBC. We also measured the fasting lipid levels baseline and after 8 yrs of an AI. Differences between the two groups were assessed by the independent samples T-test and the correlation by the linear regression analysis. Results: Fifty four pts were included in the study with a mean age of 62.5 yrs(+/-8.6 yrs). Seventeen pts are too early to be evaluable, 8 pts stopped letrozole due to adverse events and 2 pts had no baseline DEXA values. Two pts received tamoxifen 2 to 5 yrs before inclusion in the SOLE study. The remaining 25 pts showed a mean change in BMD for the lumbar spine of -1.1 (10.6SD) and for the hip -4.4 (7.04SD). Currently 13 pts in the C arm demonstrated a mean decrease in BMD for the (LS) of -2.7(10.2SD), and for the hip of -2.7(5.5SD). Twelve pts included in the I arm experienced a mean increase in BMD for the LS of 0.65(SD11.2) and a mean decrease in BMD for the hip of -4.9(9.1SD). The difference between the two treatment groups was not significant for the LS measurements after 8yrs (p=0.4) nor for the hip (p=0.23) neither was it at baseline (p=0.9; 0.1). Only three pts had fractures due to trauma. The overall fracture rate was only 0.05%.There was no correlation with the BMI, but there was a significant correlation with baseline BMD.(p=0.002)The mean fasting cholesterol levels at 8yrs in the C arm was 214mg/dl(41.3SD) and in the I arm was 218mg/dl(35.6SD)and was not significantly different(p=0.8), nor was it at baseline(p=0.5). Conclusion: This first prospective long term BMD and lipid follow-up study in PM EBC pts taking adjuvant letrozole beyond 5 yrs shows a decline in BMD. Until now no significant differences were observed between continuous and intermittent letrozole. An updated and detailed follow-up on more patients will be presented. Citation Format: Christel Fontaine, Lore Decoster, Denis Schallier, Leen Vanacker, Jacques De Grève, Marian Vanhoeij, Guy Verfaillie, Jan Lamote. Prospective study of aromatase inhibitor-induced bone loss and lipid levels in early hormone sensitive breast cancer treated with AI during 8 years [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P1-12-11.

Dienogest reduces HSD17β1 expression and activity in endometriosis.

Endometriosis is an estrogen-dependent disease. Abnormally biosynthesized estrogens in endometriotic tissues induce the growth of the lesion and worsen endometriosis-associated pelvic pain. Dienogest (DNG), a selective progesterone receptor agonist, is widely used to treat endometriosis and efficiently relieves the symptoms. However, its pharmacological action remains unknown. In this study, we elucidated the effect of DNG on enzymes involved in local estrogen metabolism in endometriosis. Surgically obtained specimens of 23 ovarian endometriomas (OE) and their homologous endometrium (EE), ten OE treated with DNG (OE w/D), and 19 normal endometria without endometriosis (NE) were analyzed. Spheroid cultures of stromal cells (SCs) were treated with DNG and progesterone. The expression of aromatase, 17β-hydroxysteroid dehydrogenase 1 (HSD17β1), HSD17β2, HSD17β7, HSD17β12, steroid sulfatase (STS), and estrogen sulfotransferase (EST) was evaluated by real-time quantitative PCR. The activity and protein level of HSD17β1 were measured with an enzyme assay using radiolabeled estrogens and immunohistochemistry respectively. OESCs showed increased expression of aromatase, HSD17β1, STS, and EST, along with decreased HSD17β2 expression, when compared with stromal cells from normal endometria without endometriosis (NESCs) (P<0.01) or stromal cells from homologous endometrium (EESCs) (P<0.01). In OESCs, DNG inhibited HSD17β1 expression and enzyme activity at 10(-7) M (P<0.01). Results of immunohistochemical analysis displayed reduced HSD17β1 staining intensity in OE w/D (P<0.05). In conclusion, DNG exerts comprehensive inhibition of abnormal estrogen production through inhibition of aromatase and HSD17β1, contributing to a therapeutic effect of DNG on endometriosis.

Identification and Qualitative Analysis of β-Sitosterol and Some Phytoestrogens in In Vivo and In-Vitro Samples of Lepidium sativum: A Semi-Arid Bone Healing Plant

Dry powder of different plant parts of Lepidium sativum (Garden Cress) are known to be used for treatment of fracture (bone healing) from ancient times. Seeds as in vivo plant part and callus as in vitro plant parts were chromatographically tested to identify and estimate β-sitosterol and various phytoestrogens. Plant-derived sterols and estrogens in tissues and oilseeds of Lepidium sativum were isolated by solvent extraction method. β-sitosterol, diadzein and formononetin were estimated qualitatively and quantitatively through TLC and HPTLC techniques. In vitro grown callus was compared biochemically with in vivo plant parts. CAMAG HPTLC was used and the developed plates were photo-documented using UV and white light. The amount of β-sitosterol was estimated to be about 0.20% w/w for seed powder and 0.024% w/w for callus powder. Daidzein and formononetin isolated from the samples of Lepidium sativum showed Rf values 0.73 and 0.67 and peak areas were 546.8 and 1033.5, respectively. Chloroform: methanol (8:2) gave excellent results in the present investigation for all samples. Results indicated that applied assay is accurate and reliable for the determination of phytoestrogens in the plant.

Tetratricopeptide repeat domain 9A negatively regulates estrogen receptor alpha activity.

Tetratricopeptide repeat domain 9A (TTC9A) is a target gene of estrogen and progesterone. It is over-expressed in breast cancer. However, little is known about the physiological function of TTC9A. The objectives of this study were to establish a Ttc9a knockout mouse model and to study the consequence of Ttc9a gene inactivation. The Ttc9a targeting vector was generated by replacing the Ttc9a exon 1 with a neomycin cassette. The mice homozygous for Ttc9a exon 1 deletion appear to grow normally and are fertile. However, further characterization of the female mice revealed that Ttc9a deficiency is associated with greater body weight, bigger thymus and better mammary development in post-pubertal mice. Furthermore, Ttc9a deficient mammary gland was more responsive to estrogen treatment with greater mammary ductal lengthening, ductal branching and estrogen target gene induction. Since Ttc9a is induced by estrogen in estrogen target tissues, these results suggest that Ttc9a is a negative regulator of estrogen function through a negative feedback mechanism. This is supported by in vitro evidence that TTC9A over-expression attenuated ERα activity in MCF-7 cells. Although TTC9A does not bind to ERα or its chaperone protein Hsp90 directly, TTC9A strongly interacts with FKBP38 and FKBP51, both of which interact with ERα and Hsp90 and modulate ERα activity. It is plausible therefore that TTC9A negatively regulates ERα activity through interacting with co-chaperone proteins such as FKBP38 and FKBP51.

Expression of estrogen and androgen receptors in tissues of external genitalia of girls with congenital adrenal hyperplasia

The reason of post-operative introital stenosis in girls with CAH is a poor estrogenization of external genitalia before vaginoplasty. It is possible that the local sensitivity to estrogens can be reduced due to prenatal androgen excess in addition to inadequate compensation, which results to decrease of estrogens production by the ovaries. Objective. To determine the distribution of estrogen and androgen receptors in genital tissue, depending on the form of CAH, the degree of external virilization and serum levels of androgens. Material and methods. The waste surgical tissues obtained during routine feminizing genitoplasty (clitoro- and vaginoplasty) was used for immunohistochemical study of estrogen (α and β) and androgen receptors in girls with CAH (n=13 and n=8, respectively). Results. The expression of estrogen (α, β) and androgen receptors was determined in genital tissues of girls with CAH. There was no significant correlation between the number of immunopositive cells in the tissues of labia minora and vagina and the degree of virilization, as well as between the number of cells expressing the receptors and serum levels of androgens (testosterone, 17-hydroxyprogesterone). There was no difference in the expression of estrogen and androgen receptors between girls with salt-wasting and simple virilizing forms of CAH. The expression of estrogen receptor α in vaginal tissues was reduced in girls who underwent repeated vaginoplasty and were under preoperative local estrogen therapy in comparison to patients in whom the vaginoplasty was performed for the first time (p=0.045). Conclusion. The distribution of estrogen and androgen receptors in the labial and vaginal tissues in girls with CAH doesn’t depend on the form of disease, external virilization and serum levels of androgens. Girls undergoing repeated vaginoplasty may have a reduced sensitivity to local estrogen therapy due to decrease of estrogen receptors expression in the vaginal mucosa.

Hormone responsive breast cancer and BRCA1 mutation: mechanism, regulation and iron-mediated effects.

Breast cancer is a prominent cause of mortality in women worldwide, with about 2/3(rd) cases linked to hormone mediated malignancy itself. A hormone receptor positive breast cancer represents cells showing rigorous proliferation upon hormonal exposure. BRCA1 is the predominant marker gene responsible for estrogen regulation. However increased exposure to estrogen is not the sole cause for this abnormality as there is no significant alteration reported in breast tissue estrogen levels. Iron metabolism has also been shown to be frequently altered in breast cancer with considerably higher iron in post menstrual women. In fact estrogen and iron have been implicated to exert synergistic effects on cellular proliferation in BRCA1 linked hormone responsive breast cancer. Thus establishing a link between estrogen and iron metabolism has a great prognostic value in predicting clinical outcome in BRCA1 linked hormone responsive breast cancer patients. Since the time immemorial Iron chelators have been implicated in combating iron dysregulation especially in breast cancer. We summarize here in this review the recent advancements in the area of iron chelation therapy delineating the role of iron in hormone receptor positive Breast cancer.

Estrogen receptor β: the guardian of the endometrium.

The endometrium is the primary target organ for the 'female' sex steroid hormone estrogen, which exerts effects in the endometrium via two main classical estrogen receptor (ER) isoforms, ERα and ERβ. The main function of the endometrium, embryo implantation, appears unperturbed in ERβ knockout mice, which has led researchers to disregard other potentially important functional roles that ERβ may have in endometrium. This review focuses on ERβ in the human endometrium and its protective role from the undesired effects of ERα. We conducted a systematic search using PubMed and Ovid for publications between January 1996 and February 2014. All studies that examined ERβ expression or function in non-pregnant endometrium or cells derived from the endometrium were considered, including human and animal studies. Studies of the basic function of ERβ isoforms in restraining ERα-mediated cell-specific trophic/mitotic responses to estrogen in other tissues has allowed appreciation of the important potential role of ERβ in the regulation of cell fate in the human endometrium. Our current understanding of ERβ expression and function in endometrium is, however, incomplete. ERβ is dynamically expressed in healthy premenstrual endometrium, persists in post-menopausal atrophic endometrium and may play an important role in endometrial disease. All endometrial cell types express ERβ and aberrations in ERβ expression have been reported in almost all benign and malignant endometrial proliferative disease. The collective evidence suggests that ERβ has an important role in normal endometrial function and also in most, if not all, benign and malignant endometrial diseases. However, the conduct of studies of endometrial ERβ expression needs to be standardized: agreement is needed regarding the most appropriate control tissue for endometrial cancer studies as well as development of standardized methods for the quantification of ERβ immunohistochemical data, similar to those scoring systems employed for other hormonally regulated tissues such as breast cancer, since these data may have direct clinical implications in guiding therapy.

Expression of PTHrP and PTH/PTHrP receptor 1 in the superior cervical ganglia of rats

PTHrP and its receptor PTHR1 are found in the CNS and peripheral nervous system. The presence of PTHrP mRNA has been detected in the superior cervical ganglion (SCG), but there are no data on the cellular distribution of PTHrP and PTHR1 in the SCG. Although it is known that ovarian activity and reproductive status influence sympathetic activity, and the PTHrP/PTHR1 system is influenced by estrogens in different tissues, it is not known whether these factors have a similar effect on expression of PTHrP and PTHR1 in the nervous system. Hence, we investigated the presence and distribution of PTHrP and PTHR1 in neurons and glia of the SCG of rats, as well as the influence of ovariectomy on their expression, by using immunohistochemistry. PTHrP and PTHR1 immunoreactivity was observed in cytoplasm as well as in nuclei of almost all neurons in the SCG. In male rats, intensity of PTHrP fluorescence was significantly higher in cytoplasm of NPY−, in comparison to NPY+ neurons (p < 0.05). In female rats, 2 months post-ovariectomy, significantly lower intensity of PTHrP fluorescence in cytoplasm of the SCG neurons was observed in comparison to sham operated animals (p < 0.05). In addition to neurons, PTHrP and PTHR1 immunoreactivity was observed in most of the glia and was not influenced by ovariectomy. Results show the expression of PTHrP and its receptor, PTHR1, in the majority of neurons and glial cells in the SCG of rats. Expression of PTHrP, but not PTHR1 in the cytoplasm of SCG neurons is influenced by ovarian activity.