estrone-sulfate and Endometrial-Neoplasms

The identification of genes and pathways that are affected by estrogenization may shed light on the mechanisms of estrogen action. Here, we describe the expression pattern of a novel estrogen-induced gene, EIG121, in distinct types of endometrial cancer.. EIG121 was identified by cDNA microarray analysis of endometrial RNA from women receiving either placebo or estrogen replacement therapy. The expression level of EIG121 was then measured by real-time quantitative reverse transcription-PCR in benign, hyperplastic, and malignant endometrial samples. A polyclonal antibody was used to detect EIG121 protein by immunohistochemistry.. In postmenopausal endometrium, estrogen replacement therapy with Premarin and synthetic estrogen sulfate conjugates induced the expression of EIG121 2- and 3-fold, respectively. In premenopausal endometrium, the expression of EIG121 was higher in the estrogen-dominated proliferative phase than the secretory phase. In endometrial complex, hyperplasia, and endometrioid adenocarcinoma, neoplastic proliferations associated with estrogen excess, the expression of EIG121 was significantly elevated (on average 3.8-fold in hyperplasias and 21-fold in grade 1 tumors). Although the level of EIG121 mRNA in grade 3 endometrioid carcinoma was still 3.5-fold of that in benign endometrium, EIG121 expression tended to decline with increasing tumor grade and disease stage. Immunohistochemistry showed faint staining of normal endometrial epithelium, but intense staining of endometrioid tumors. In sharp contrast, EIG121 expression was significantly suppressed in both uterine papillary serous carcinoma and uterine malignant mixed mullerian tumor, two tumors not associated with estrogen exposure, to <5% of the level in benign endometrium.. Our results suggest that EIG121 is a good endometrial biomarker associated with a hyperestrogenic state and estrogen-related type I endometrial adenocarcinoma.

Topics: Adenocarcinoma; Biomarkers, Tumor; Case-Control Studies; Endometrial Hyperplasia; Endometrial Neoplasms; Estrogen Replacement Therapy; Estrogens; Estrogens, Conjugated (USP); Estrone; Expressed Sequence Tags; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Membrane Proteins; Neoplasm Proteins; Oligonucleotide Array Sequence Analysis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Endometrial cancer (EC) is associated with increased estrogen actions. Locally, estrogens can be formed from estrone-sulphate (E1-S) after cellular uptake by organic anion-transporting polypeptides (OATP) or organic anion transporters (OAT). Efflux of E1-S is enabled by ATP Binding Cassette transporters (ABC) and organic solute transporter (OST)αβ. Currently, 19 E1-S transporters are known but their roles in EC are not yet understood. Here, we analysed levels of E1-S transporters in Ishikawa (premenopausal EC), HEC-1-A (postmenopausal EC), HIEEC (control) cell lines, in EC tissue, examined metabolism of steroid precursor E1-S, studied effects of OATPs' inhibition and gene-silencing on E1-S uptake, and assessed associations between transporters and histopathological data. Results revealed enhanced E1-S metabolism in HEC-1-A versus Ishikawa which could be explained by higher levels of OATPs in HEC-1-A versus Ishikawa, especially 6.3-fold up-regulation of OATP1B3 (

Topics: Age Factors; ATP Binding Cassette Transporter, Subfamily G, Member 2; Biological Transport; Cell Line, Tumor; Endometrial Neoplasms; Estrone; Female; Fluorescent Antibody Technique; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Membrane Transport Proteins; Multigene Family; Neoplasm Grading; Neoplasm Invasiveness; Neoplasm Proteins; Neoplasm Staging; Organic Anion Transporters; Postmenopause; Solute Carrier Organic Anion Transporter Family Member 1B3

Estrogens have important roles in the pathogenesis of endometrial cancer. They can have carcinogenic effects through stimulation of cell proliferation or formation of DNA-damaging species. To characterize model cell lines of endometrial cancer, we determined the expression profiles of the estrogen receptors (ERs) ESR1, ESR2 and GPER, and 23 estrogen biosynthetic and metabolic genes, and investigated estrogen biosynthesis in the control HIEEC cell line and the Ishikawa and HEC-1A EC cell lines. HIEEC and Ishikawa expressed all ERs to different extents, while HEC-1A cells lacked expression of ESR1. Considering the estrogen biosynthetic and metabolic enzymes, these cells showed statistically significant different gene expression profiles for SULT2B1, HSD3B2, CYP19A1, AKR1C3, HSD17B1, HSD17B7, HSD17B12, CYP1B1, CYP3A5, COMT, SULT1A1, GSTP1 and NQO2. In these cells, E2 was formed from E1S and E1, while androstenedione was not converted to estrogens. HIEEC and Ishikawa had similar profiles of androstenedione and E1 metabolism, but hydrolysis of E1S to E1 was weaker in Ishikawa cells. HEC-1A cells were less efficient for activation of E1 into the potent E2, but metabolized androstenedione to other androgenic metabolites better than HIEEC and Ishikawa cells. This study reveals that HIEEC, Ishikawa, and HEC-1A cells can all form estrogens only via the sulfatase pathway. HIEEC, Ishikawa, and HEC-1A cells expressed all the major genes in the production of hydroxyestrogens and estrogen quinones, and in their conjugation. Significantly higher CYP1B1 mRNA levels in Ishikawa cells compared to HEC-1A cells, together with lack of UGT2B7 expression, indicate that Ishikawa cells can accumulate more toxic estrogen-3,4-quinones than HEC-1A cells, as also for HIEEC cells. This study provides further characterization of HIEEC, Ishikawa, and HEC-1A cells, and shows that they differ greatly in expression of the genes investigated and in their capacity for E2 formation, and thus they represent different in vitro models.

Topics: 3-Hydroxysteroid Dehydrogenases; Aldo-Keto Reductase Family 1 Member C3; Androstenedione; Aromatase; Arylsulfotransferase; Catechol O-Methyltransferase; Cell Line, Tumor; Endometrial Neoplasms; Estrogens; Estrone; Female; Glutathione S-Transferase pi; Humans; Hydroxyprostaglandin Dehydrogenases; Progesterone Reductase; Quinone Reductases; Quinones; RNA, Messenger; Sulfatases; Sulfotransferases; Transcriptome

The involvement of aromatase, steroid sulfatase (STS) and reductive 17beta-hydroxysteroid dehydrogenases (17beta-HSDs) in the production of estrogens was determined in four cell lines of endometrial cancer (Ishikawa, HEC-1A, HEC-1B and RL-95) and one cell line of cervix cancer (Hela) in culture. After incubation with 4-androstene-3,17-dione (4-dione), there are no estrogens, estrone (E1) and estradiol (E2), detected suggesting that the pathway of aromatase is not important in these cell lines. In whole cells, the results show low percentages of transformation of estrone sulfate (E1S) into E1 suggesting that the entrance of E1S is difficult. However, in homogenized cells the STS activity was much higher and fully blocked by an inhibitor. Using selective inhibitors for each reductive 17beta-HSD (types 1, 5, 7 and 12), alone or in combination, we did not succeed in completely blocking the conversion of E1 into E2, suggesting that another 17beta-HSD (known or unknown) is involved in the formation of E2 from E1.

Topics: 17-Hydroxysteroid Dehydrogenases; Aromatase; Cell Line, Tumor; Endometrial Neoplasms; Estradiol; Estrogens; Estrone; Female; Humans; Isoenzymes; Steryl-Sulfatase; Uterine Cervical Neoplasms

Identification of biomarkers potentially provides prognostic information that can help guide clinical decision-making. Given the relationship between estrogen exposure and endometrial cancer, especially low grade endometrioid carcinoma, we hypothesized that high expression of genes induced by estrogen would identify low risk endometrioid endometrial cancers. cDNA microarray and qRT-PCR verification were used to identify six genes that are highly induced by estrogen in the endometrium. These estrogen-induced biomarkers were quantified in 72 endometrial carcinomas by qRT-PCR. Unsupervised cluster analysis was performed, with expression data correlated to tumor characteristics. Time to recurrence by cluster was analyzed using the Kaplan-Meier method. A receiver operating characteristic (ROC) curve was generated to determine the potential clinical utility of the biomarker panel to predict prognosis. Expression of all genes was higher in endometrioid carcinomas compared to non-endometrioid carcinomas. Unsupervised cluster analysis revealed two distinct groups based on gene expression. The high expression cluster was characterized by lower age, higher BMI, and low grade endometrioid histology. The low expression cluster had a recurrence rate 4.35 times higher than the high expression cluster. ROC analysis allowed for the prediction of stage and grade with a false negative rate of 4.8% based on level of gene expression in endometrioid tumors. We have therefore identified a panel of estrogen-induced genes that have potential utility in predicting endometrial cancer stage and recurrence risk. This proof-of-concept study demonstrates that biomarker analysis may play a role in clinical decision making for the therapy of women with endometrial cancer.

Topics: Adult; Age Factors; Aged; Aged, 80 and over; Biomarkers, Tumor; Body Mass Index; Carcinoma, Endometrioid; Cluster Analysis; Endometrial Neoplasms; Equilin; Estrogen Replacement Therapy; Estrogens, Conjugated (USP); Estrone; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Genetic Association Studies; Humans; Middle Aged; Neoplasm Proteins; Oligonucleotide Array Sequence Analysis; Prognosis; Randomized Controlled Trials as Topic; Reverse Transcriptase Polymerase Chain Reaction; ROC Curve

Data suggest that post-menopausal women with larger ovaries are at increased risk for endometrial carcinoma; however, analyses comparing ovarian volume to serum hormone levels are limited. Accordingly, we assessed ovarian volumes in relation to serum sex hormone levels among post-menopausal women with endometrial carcinoma who participated in a multi-center case-control study.. Data for established risk and protective factors for endometrial carcinoma were collected via in-person interviews. Ovarian volumes were estimated from pathology reports. Associations between exposures and age-adjusted ovarian volumes were analyzed for 175 cases with available data. For a subset of 135 cases, we analyzed relationships between ovarian volume, adjusted for age and body mass index (BMI), and serum hormone levels by analysis of variance.. Ovarian volume declined progressively from 1.83 cm3 among women ages 55-59 years to 1.23 cm3 among women age 70 years or older (p-trend=0.02). Larger ovarian volume was associated with early menarche (p-trend=0.03), having given birth (p=0.01), and weakly with elevated BMI (p-trend=0.06). After adjustment, increased ovarian volume was associated with higher estradiol (p-trend=0.007); albumin-bound estradiol (p-trend=0.01); and free estradiol (p-trend=0.006) levels; androstenedione, estrone and estrone sulfate showed similar, though non-significant associations.. Among women with endometrial carcinoma, larger ovaries were associated with higher serum levels of estrogens. Further studies examining the role of the ovaries in post-menopausal hormonal carcinogenesis are warranted.

Topics: Aged; Androstenedione; Case-Control Studies; Endometrial Neoplasms; Estradiol; Estrone; Female; Gonadal Steroid Hormones; Humans; Middle Aged; Ovary; Postmenopause; Sex Hormone-Binding Globulin

Data on activities of estrogen-synthesis enzymes (aromatase and steroidsulfatase) in endometrial tumors were obtained by sensitive and highly precise radioisotope methods. The results clear out some pathogenetic aspects in the development and progress of malignant tumors in the endometrium.

Topics: Aromatase; Cell Differentiation; Endometrial Neoplasms; Estrone; Female; Humans; Isotope Labeling; Menopause; Middle Aged; Neoplasm Staging; Postmenopause; Steryl-Sulfatase; Tritium

The present studies concern sulphotransferase activities for estrogens and other steroids, and 17beta-hydroxysteroid dehydrogenase (17beta-HSD) activities for estrogens in Ishikawa endometrial adenocarcinoma cells. When physiological concentrations of various estrogens (estrone, estradiol, estriol) are incubated, most of the transformation product is the respective sulphate. The sulphotransferase activity is very rapid, and 2 h after incubation 70-95% are converted to the sulphated form. Sulphates are found exclusively in the culture medium, which suggests that as soon as the sulphate is biosynthesized it is secreted to the medium. Comparative data using neutral steroids (dehydroepiandrosterone, testosterone, and pregnenolone) show that sulphotransferase activity for these compounds is very limited. In another series of studies, 17beta-HSD activity was explored for the interconversion estrone estradiol. At low concentrations (5 x 10(-9)-5 x 10(-8) M), when estradiol (E2) is incubated, most of the unconjugated material remains as E2 in the cellular compartment, but at high concentrations (5 x 10(-7)-5 x 10(-6) M) a great proportion (70-80%) of the E2 is converted to estrone (E1). On the other hand, after incubation of E1 at all concentrations most remained as unchanged E1. It is suggested that, in Ishikawa cells, at very low concentrations of E1 or E2, sulphotransferases are predominant, but when this enzyme is saturated 17beta-HSD activity is orientated to the oxidative form.

Topics: 17-Hydroxysteroid Dehydrogenases; Adenocarcinoma; Dehydroepiandrosterone; Endometrial Neoplasms; Estrogens; Estrone; Female; Humans; Pregnenolone; Sulfotransferases; Testosterone; Tumor Cells, Cultured

It has been suggested that identified risk factors for endometrial cancer operate through a single etiologic pathway, i.e., exposure to relatively high levels of unopposed estrogen (estrogen in the absence of progestins). Only a few studies, however, have addressed this issue directly.. We assessed the risk of developing endometrial cancer among both premenopausal and postmenopausal women in relation to the circulating levels of steroid hormones and sex hormone-binding globulin (SHBG). The independent effect of hormones was assessed after adjustment for other known risk factors.. The data used in the analysis are from a case-control study conducted in five geographic regions in the United States. Incident cases were newly diagnosed during the period from June 1, 1987, through May 15, 1990. The case patients, aged 20-74 years, were matched to control subjects by age, race, and geographic region. The community control subjects were obtained by random-digit-dialing procedures (for subjects 20-64 years old) and from files of the Health Care Financing Administration (for subjects > or = 65 years old). Additional control subjects who were having a hysterectomy performed for benign conditions were obtained from the participating centers. Women reporting use of exogenous estrogens or oral contraceptives within 6 months of interview were excluded, resulting in 68 case patients and 107 control subjects among premenopausal women and 208 case patients and 209 control subjects among postmenopausal women. The hormone analyses were performed on blood samples obtained from case patients or from hysterectomy control subjects before surgery. The odds ratios (ORs) and 95% confidence intervals (CIs) were estimated by use of an unconditional logistic regression analysis after we controlled for matching variables and potential confounders. All P values were two-sided.. High circulating levels of androstenedione were associated with 3.6-fold and 2.8-fold increased risks among premenopausal and postmenopausal women, respectively, after adjustment for other factors (P for trend = .01 and < .001, respectively). Risks related to other hormone fractions varied by menopausal status. Among postmenopausal women, a reduced risk was associated with high SHBG levels and persisted after adjustment was made for obesity and other factors (OR = 0.51; 95% CI = 0.27-0.95). High estrone levels were associated with increased risk (OR = 3.8; 95% CI = 2.2-6.6), although adjustment for other risk factors (particularly body mass index) diminished the effect (OR = 2.2; 95% CI = 1.2-4.4). Albumin-bound estradiol (E2), a marker of the bioavailable fraction, also remained an important risk factor after adjustment was made for other factors (OR = 2.0; 95% CI = 1.0-3.9). In contrast, high concentrations of total, free, and albumin-bound E2 were unrelated to increased risk in premenopausal women. In both premenopausal and postmenopausal groups, risks associated with obesity and fat distribution were not affected by adjustment for hormones.. High endogenous levels of unopposed estrogen are related to increased risk of endometrial cancer, but their independence from other risk factors is inconsistent with being a common underlying biologic pathway through which all risk factors for endometrial cancer operate.. Further research should focus on alternative endocrinologic mechanisms for risk associated with obesity and body fat distribution and for the biologic relevance of the increased risk associated with androstenedione in both premenopausal and postmenopausal disease.

Topics: Adult; Androstenedione; Case-Control Studies; Endometrial Neoplasms; Estradiol; Estrogens, Conjugated (USP); Estrone; Female; Gonadal Steroid Hormones; Humans; Middle Aged; Odds Ratio; Postmenopause; Premenopause; Reproducibility of Results; Risk; Risk Factors; Sex Hormone-Binding Globulin; Single-Blind Method

The metabolism of [3H]estrone sulfate ([3H]E1S) was studied in normal breast tissue from 10 premenopausal women without oral contraceptives (OC), in 12 OC users and in 9 untreated postmenopausal women. [3H]E1S was converted into estrone ([3H]E1) and estradiol-17 beta ([3H]E2) by tissue samples from all three groups of women, with only minor formation of other unconjugated compounds. The rate of [3H]E2 formation was significantly higher in premenopausal women without OC than in postmenopausal women. Among premenopausal women, OC users had a significantly lower rate of total hydrolysis and of [3H]E1 formation than non-users. The rate of total hydrolysis of [3H]E1S in normal breast tissue from all three groups of women was similar to that in muscle, but the rate of [3H]E2 formation was ten times higher. Both total hydrolysis rate and rate of [3H]E2 formation were significantly lower in normal breast tissue than in breast carcinoma and in normal and neoplastic endometrium. The specific ability of normal breast tissue to convert E1S into the terminal biologically active estrogen E2 may be important for estrogenic stimulation of the breast in subjects with low circulating E2 levels. The lower rate of E1 formation in OC users may reflect an inhibitory effect of the progestagen compound in such preparations.

Topics: Adolescent; Adult; Aged; Breast; Breast Neoplasms; Contraceptives, Oral; Endometrial Neoplasms; Estradiol; Estrone; Female; Humans; Menopause; Middle Aged; Postmenopause; Premenopause; Progesterone; Tritium

Estrone sulfate (E1-S) has been shown to be quantitatively the most important estrogen in peripheral blood. But, the physiological and/or pathological role of E1-S is not yet clarified. At present, we tried to clarify it using tissue cultures. In tissue cultures of human endometrium, secretory endometrium showed higher activity of estrone sulfatase (E1----E1-S) than proliferative endometrium. Progesterone added in the medium induced an increase of estrone sulfotransferase in the proliferative endometrium. The results suggest a reducing effect of estrogen by progesterone in secretory endometrium in physiological conditions. Estrogen dependent malignant tumors (breast cancer, endometrial cancer) have high estrone sulfatase. It converts E1-S to E1 (----E2) which are abundant in these tumors. Ishikawa cell line increased estrone sulfotransferase activity with progesterone, somewhat like the physiological conditions. From out study in vivo, there is a possibility of some ameliorative effects of E1-S on the central nervous system of patients with senile dementia (Alzheimer's type). Effects of E1-S on central nerves were investigated using tissue cultures.

Topics: Alzheimer Disease; Breast Neoplasms; Culture Techniques; Endometrial Neoplasms; Endometrium; Estradiol Dehydrogenases; Estrone; Female; Humans; Progesterone; Sulfatases; Sulfotransferases; Sulfurtransferases