zearalenone and Endometrial-Neoplasms

Endometrial cancer is one of the most commonly diagnosed cancers in women. The search for factors that contribute to the development of cancer cells in reproductive organs should involve the detection of xenoestrogens, in particular zearalenone (ZEA) and its metabolites. Xenoestrogens are endocrine disruptors-ZEA and its metabolites are structurally similar to estrogens (macrocyclic lactone ring) and show high affinity for estrogen receptors. This study proposes a new method for the preparation of samples of human tissues with endometrial cancer by the use of the QuEChERS technique. Analytical parameters such as centrifugation temperature, extraction solvent, and adsorbents were modified to obtain satisfactory recovery for ZEA (R = 82.6%, RSD = 2.9%) and one of its metabolites, α-zearalenol (R = 50.1%, RSD = 3.2%). High-performance liquid chromatography (HPLC) with fluorescence detection and tandem mass spectrometry (LC-QTOF-MS) were used for the identification and quantitative determination of the analyzed compounds. The developed procedure was applied for analyses of human tissues with endometrial cancer. The presence of α-zearalenol was detected in 47 out of the 61 examined tissue samples. Graphical Abstract Methodology for isolation and identification of zearalenone and its major metabolites.

Topics: Chromatography, High Pressure Liquid; Endometrial Neoplasms; Female; Humans; Tandem Mass Spectrometry; Zearalenone

This study presents a selective method of isolation of zearalenone (ZON) and its metabolite, α-zearalenol (α-ZOL), in neoplastically changed human tissue by accelerated solvent and ultrasonic extractions using a mixture of acetonitrile/water (84/16% v/v) as the extraction solvent. Extraction effectiveness was determined through the selection of parameters (composition of the solvent mixture, temperature, pressure, number of cycles) with tissue contamination at the level of nanograms per gram. The produced acetonitrile/water extracts were purified, and analytes were enriched in columns packed with homemade molecularly imprinted polymers. Purified extracts were determined by liquid chromatography (LC) coupled with different detection systems (diode array detection--DAD and mass spectrometry--MS) involving the Ascentis RP-Amide as a stationary phase and gradient elution. The combination of UE-MISPE-LC (ultrasonic extraction--molecularly imprinted solid-phase extraction--liquid chromatography) produced high (R≈95-98%) and repeatable (RSD<3%) recovery values for ZON and α-ZOL.

Topics: Aged; Chromatography, Liquid; Endometrial Neoplasms; Female; Humans; Mass Spectrometry; Polymers; Solid Phase Extraction; Zearalenone; Zeranol

Several anthropogenous and naturally occurring substances, referred to as estrogen active compounds (EACs), are able to interfere with hormone and in particular estrogen receptor signaling. EACs can either cause adverse health effects in humans and wildlife populations or have beneficial effects on estrogen-dependent diseases. The aim of this study was to examine global gene expression profiles in estrogen receptor (ER)-proficient Ishikawa plus and ER-deficient Ishikawa minus endometrial cancer cells treated with selected well-known EACs (diethylstilbestrol, genistein, zearalenone, resveratrol, bisphenol A and o,p'-DDT). We also investigated the effect of the pure antiestrogen ICI 182,780 (ICI) on the expression patterns caused by these compounds. Transcript levels were quantified 24 h after compound treatment using Illumina BeadChip Arrays. We identified 87 genes with similar expression changes in response to all EAC treatments in Ishikawa plus. ICI lowered the magnitude or reversed the expression of these genes, indicating ER dependent regulation. Apart from estrogenic gene regulation, bisphenol A, o,p'-DDT, zearalenone, genistein and resveratrol displayed similarities to ICI in their expression patterns, suggesting mixed estrogenic/antiestrogenic properties. In particular, the predominant antiestrogenic expression response of resveratrol could be clearly distinguished from the other test compounds, indicating a distinct mechanism of action. Divergent gene expression patterns of the phytoestrogens, as well as weaker estrogenic gene expression regulation determined for the anthropogenous chemicals bisphenol A and o,p'-DDT, warrants a careful assessment of potential detrimental and/or beneficial effects of EACs. The characteristic expression fingerprints and the identified subset of putative marker genes can be used for screening chemicals with an unknown mode of action and for predicting their potential to exert endocrine disrupting effects.

Topics: Benzhydryl Compounds; Cell Line, Tumor; Cluster Analysis; DDT; Diethylstilbestrol; Endocrine Disruptors; Endometrial Neoplasms; Estradiol; Estrogen Antagonists; Estrogens; Female; Fulvestrant; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Genistein; Humans; Oligonucleotide Array Sequence Analysis; Phenols; Phytoestrogens; Polymerase Chain Reaction; Receptors, Estrogen; Reproducibility of Results; Resveratrol; Risk Assessment; RNA, Messenger; Stilbenes; Time Factors; Zearalenone

The glycoprotein Wnt-7a participates in a signaling pathway that transmits information among uterine cell types. Disruption of this pathway by the transplacentally acting carcinogen diethylstilbestrol (DES) is associated with morphological abnormalities of the female reproductive tract (FRT). This raises the question whether estrogens in the diet might also interfere with this pathway. Therefore, this study investigated the influence of the steroid hormone 17beta-estradiol (E2), the mycotoxin zearalenone (ZEN), the soy phytoestrogen genistein (GEN), and DES on the expression of Wnt-7a in an endometrial adenocarcinoma cell line (Ishikawa cells) by reverse transcription/competitive PCR. In addition, the enzymatic activity of alkaline phosphatase (ALP) was determined, which is estrogen receptor (ER)-dependently regulated in Ishikawa cells. After treatment of Ishikawa cells with E2, ZEN, GEN, and DES, a decrease in the gene expression of Wnt-7a was observed. Maximum effect (50% reduction) was observed after treatment with concentrations that induced maximum expression of the ALP. Experiments in the presence of the ER antagonist (ICI 182,780) suggested that the ER is involved in the regulation of Wnt-7a in Ishikawa cells. In conclusion, interference with the expression of Wnt genes in the FRT might be a novel mechanism by which estrogens disrupt the function of the FRT.

Topics: Adenocarcinoma; Alkaline Phosphatase; Cell Line, Tumor; Diethylstilbestrol; Endometrial Neoplasms; Endometrium; Estradiol; Estrogen Receptor beta; Estrogens; Female; Gene Expression; Genistein; Humans; Proto-Oncogene Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Wnt Proteins; Zearalenone

Zearalenone (ZEA) is a nonsteroidal estrogenic compound mainly produced by the molds Fusarium graminearium and Fusarium culmorum found in a variety of host plants and soil debris around the world. ZEA is usualy non-lethal to animals but is important to livestock producers because its hyperestrogenic effects adversely influence the reproductive performance of animals. There have been suggestions of possible involvement of ZEA in the progression of breast malignancies and tumors of the female reproductive tract in humans. The toxic or stimulatory effects of ZEA and its metabolites alpha-zearalenol and 17-beta-estradiol on SKN, HHUAand HepG2 cells were studied using rapid colorimetric MTT assay. In general, both concentrations of 17-beta-estradiol (100M and 10 nM) were toxic to SKN and HHUA cell cultures. Both ZEA and alpha-zearalenol stimulated the proliferation of SKN and HHUA cells. On HepG2 cells, lower concentrations (10 nM) of 17-beta-estradiol and higher concentrations (100 microM) of ZEA exhibited toxic effects, whereas treatment with higher concentrations of 17-beta-estradiol and lower concentration of ZEA did not show toxic effects. A dose dependent antagonistic effect was observed when the cell cultures were pre-incubated with ICI 182,780, a synthetic estrogen receptor blocker, before estradiol or mycotoxin treatments.

Topics: Adenocarcinoma; Carcinoma, Hepatocellular; Cell Division; Colorimetry; Dose-Response Relationship, Drug; Endometrial Neoplasms; Estradiol; Estrogens, Non-Steroidal; Female; Humans; Leiomyosarcoma; Tumor Cells, Cultured; Zearalenone; Zeranol

Zearalenone (ZEA), a nonsteroidal mycotoxin with estrogen-like activity, is synthesized by molds (Fusarium) commonly contaminating poorly stored agricultural products and foodstuffs. Human ER binds ZEA and this is probable mechanism of its action, although their influence on target tissues seems to be weaker (80-160 less active) comparing to E2. Zea has been observed to possess tumor-promoting activity similar to that of estrogens and hypothetically can inducing proliferation and carcinogenesis in estrogen-dependent tissues. Nowadays, the questions are, if ZEA is present in human endometrium and whether concentrations of this mycoestrogen is associated with endometrial cell proliferation. Endometrial tissues specimens were collected from 49 women (endometrial adenocarcinoma n = 27, endometrial hyperplasia n = 11, normal proliferative endometrium n = 11). Mean tissue zearalenone concentration in 3 endometrial hyperplasia and 22 adenocarcinoma samples was 47.8 +/- +/- 6.48 and 167 +/- +/- 17.69 ng/ml respectively in contrary to normal endometrium where tissue mycoestrogen concentration was not detectable. In 8 cases of hyperplastic and 5 cases of neoplastic endometrial tissue specimens ZEA was not observed. Our findings confirm the presence of ZEA in hyperplastic and neoplastic endometrium and therefore this substance might be of importance in carcinogenesis.

Topics: Adenocarcinoma; Adult; Aged; Biopsy; Endometrial Hyperplasia; Endometrial Neoplasms; Endometrium; Estrogens, Non-Steroidal; Female; Humans; Middle Aged; Retrospective Studies; Zearalenone