Glyphosate-based herbicide worsens alterations induced by cafeteria diet on rat uterus.

Exposure to glyphosate-based herbicides (GBH) and consumption of cafeteria (CAF) diet, which are widespread in Western society, seem to be associated with endometrial hyperplasia (EH). Here, we aimed to evaluate the effects of a subchronic low dose of GBH added to the CAF diet on the rat uterus. Female Wistar rats were fed from postnatal day (PND)21 until PND240 with chow (control) or CAF diet. Since PND140, rats also received GBH (2 mg of glyphosate/kg/day) or water through food, yielding four experimental groups: control, CAF, GBH, and CAF+GBH. On PND240, CAF and CAF+GBH animals showed an increased adiposity index. With respect to the control group, no changes in the serum levels of 17ß-estradiol and progesterone were found. However, progesterone levels were higher in the CAF+GBH group than in the CAF and GBH groups. In the uterus, both studied factors alone and in combination induced morphological and molecular changes associated with EH. Furthermore, the addition of GBH provoked an increased thickness of subepithelial stroma in rats fed with the CAF diet. As a consequence of GBH exposure, CAF+GBH rats exhibited an increased density of abnormal gland area, considered preneoplastic lesions, as well as a reduced PTEN and p27 expression, both tumor suppressor molecules that inhibit cell proliferation, with respect to control rats. These results indicate that the addition of GBH exacerbates the CAF effects on uterine lesions and that the PTEN/p27 signaling pathway seems to be involved. Further studies focusing on the interaction between unhealthy diets and environmental chemicals should be encouraged to better understand uterine pathologies.

Expression of p53 protein in the endometrial polyp in postmenopausal women.

OBJECTIVE: To evaluate p53 protein expression in the endometrial polyp and compare with adenocarcinoma and atrophic endometrium of postmenopausal women. MATERIALS AND METHODS: Ninety-eight postmenopausal women were included in this study and divided into three groups related to histopathologic diagnosis: Group A--endometrial adenocarcinoma (n = 40), Group B--endometrial polyp (n = 38), and Group C--endometrial atrophy (n = 20). The length of this study was from 1990 to 2004. The endometrial samples were collected from hysteroscopic biopsy or surgery then processed for histopathologic routine. One thousand cells of each histological section were evaluated for immunohistochemical analysis using p53 antibodies. The ANOVA test was performed for the statistical analysis. RESULTS: The expression of p53 in adenocarcinoma samples was the highest. The expression of polyp was positive when associated to hyperplasia without atypia. All samples of atrophic endometrial were negative. CONCLUSIONS: The present data suggested that presence of hyperplasia in the endometrial polyp is factor to increase the expression of p53.

Involvement of Akt, Ras and cell cycle regulators in the potential development of endometrial hyperplasia in women with polycystic ovarian syndrome.

OBJECTIVE: To examine whether the abundance, localization, and/or activity of cell cycle regulators CDK2, Cyclin E, p27, and survival proteins AKT and Ras in PCOS-associated endometria (with and without hyperplasia) differ from non-PCOS endometria. METHODS: The expression of CDK2, Cyclin E, p27, AKT and Ras was measured by immunohistochemistry and/or Western blot in 9 normal endometria (NE), 12 endometria from PCOS patients without endometrial hyperplasia (PCOSE), 7 endometria from PCOS women with endometrial hyperplasia (HPCOSE), and 9 endometria from patients with endometrial hyperplasia (HE). The activity of CDK2 was assessed by an in vitro kinase assay. RESULTS: CDK2, Cyclin E and p27 proteins were expressed mainly in the endometrial epithelial cells of the studied groups. No change in the activity of CDK2 was observed in total extracts obtained from the tissue samples. However, the nuclear expression of CDK2 in epithelial cells was slightly elevated in PCOSE and significantly increased in HPCOSE when compared to NE. Higher expression of p27 was detected in the cytoplasm of epithelial cells of PCOSE and HPCOSE when compared to NE. Also, we found an increment in Ser473-AKT phosphorylation and an over-expression of the Ras oncogene in endometria of patients with PCOS. CONCLUSION: The PCOS condition is associated with increased Ser473-AKT phosphorylation, elevated expression of Ras, increased cytoplasmic abundance of p27, and increased nuclear abundance of CDK2 in the endometrial epithelial cells. These biological events could potentially provide a chance for endometrial cells from PCOS patients to exit the controlled cell cycle and become hyperplastic at a later stage.

In situ estrogen metabolism in proliferative endometria from untreated women with polycystic ovarian syndrome with and without endometrial hyperplasia.

The aim of the present investigation was to study whether the endocrinological status of women bearing polycystic ovarian syndrome (PCOS) affects the endometrial in situ steroid metabolism. For this purpose, we evaluated the mRNA levels (RT-PCR), and the activity of steroid metabolic enzymes: P450 aromatase, steroid sulfatase (STS), estrogen sulfotransferase (EST) and 17beta-hydroxysteroid dehydrogenase (17beta-HSD) in 23 samples of normal endometria (CE), 18 PCOS endometria without treatment (PCOSE), 10 specimens from PCOS women with endometrial hyperplasia (HPCOSE), and 7 endometria from patients with endometrial hyperplasia not associated to PCOS (EH). The data showed lower levels of STS mRNA for PCOSE and HPCOSE (p<0.05, p<0.01, respectively) and of EST for HPCOSE and EH compared to control (p<0.05). However, higher levels for EST mRNA were obtained in PCOSE (p<0.05) versus CE. The mRNA and protein levels for P450 aromatase were undetectable in all analyzed endometria. The relationship between the activities of STS and EST was lower in PCOSE and HPCOSE (p<0.05) versus CE. The ratio between the mRNA from 17beta-HSD type 1/type 2 was higher in PCOSE (p<0.05), whereas, a diminution in the 17beta-HSD type 2 activity was observed in PCOSE (p<0.05). These results indicate that the activity of enzymes related to the steroid metabolism in analyzed PCOSE differ from those found in the CE. Consequently, PCOSE may present an in situ deregulation of the steroid metabolism.

Deregulation of tissue homeostasis in endometria from patients with polycystic ovarian syndrome with and without endometrial hyperplasia.

OBJECTIVE: To study the proteins involved in endometrial homeostasis in PCOS women. METHODS: Protein expression of Ki67, Bcl-2, Bax, Pro-Caspase-3 and Caspase-3 by immunohistochemistry and/or Western blot, and DNA fragmentation using in situ 3'-end labeling of apoptotic cells, was measured in 9 samples of normal endometrium (NE), 12 PCOS endometria without treatment (PCOSE), 7 endometria from PCOS women with endometrial hyperplasia (HPCOSE) and 9 endometria from patients with endometrial hyperplasia (HE). RESULTS: Cell proliferation was higher in epithelium from PCOSE (P<0.05), HPCOSE and HE vs NE. A higher Bcl-2/Bax relative ratio in PCOSE and HPCOSE was observed, in absence of active Caspase-3 and scarce DNA fragmentation in the four groups of endometria studied. CONCLUSION: As the apoptosis was scarce in all of the groups studied, endometrial homeostasis deregulation in PCOS could be a result of increased proliferation. Therefore, the onset of endometrial hyperplasia in PCOS endometrium could be linked to inadequate cell proliferation, and concomitantly to inadequate cell survival.

Androgen and estrogen receptors and co-regulators levels in endometria from patients with polycystic ovarian syndrome with and without endometrial hyperplasia.

OBJECTIVE: To study if the endocrinological status of PCOS women affects the endometrial sensitivity to steroids by evaluating the expression of androgen receptor (AR), estrogen receptor alpha (ERalpha), estrogen receptor beta (ERbeta), co-activators AIB1 and ARA70, and co-repressor NCoR. METHODS: Gene and/or protein expression of steroid receptors and co-regulators was measured in 17 samples of normal endometrium (NE), 23 PCOS endometrium without treatment (PCOSE), 11 endometria from PCOS women and with endometrial hyperplasia (HPCOSE), and 10 endometria from patients with endometrial hyperplasia (HE), using RT-PCR and/or immunohistochemistry and Western blot. RESULTS: Gene and protein expression of AR was relatively elevated in PCOSE and HPCOSE compared with NE. A significant increase in ERalpha protein expression was observed in PCOSE, preferentially in the nucleus of endometrial cells, whereas ERbeta gene and protein expression increased gradually from PCOSE to HPCOSE and HE, mainly in the epithelial compartment. Importantly, we found a gradual increase in the ERbeta/ERalpha gene and protein expression ratio in endometria from the four groups of women. AIB1 showed increased nuclear protein expression in PCOSE compared to NE, in the presence of a high expression of ARA70 in all groups. High expression of ARA70 together with a normal expression level of AIB1 was observed in HPCOSE. The cytoplasmic immunostaining of NCoR was similar between the four groups of patients. CONCLUSION: The PCOS endometrium exhibits a higher sensitivity to steroid action. We can inferred that these alterations could deregulate the transcription of genes involved in the cell cycle, which may lead to the development of endometrial hyperplasia in PCOS women.

P53 expression in spontaneous and estradiol-induced endometrial hyperplasia during menopause.

OBJECTIVE: To determine the percentage of endometrial hyperplasia positive for p53 expression in both spontaneously occurring cases or following the use of unopposed estradiol. METHODS: Fifty-four postmenopausal patients with endometrial hyperplasia diagnosed by endometrial biopsy and hysteroscopy were recruited to this study. Thirty-three patients had used unopposed estradiol for periods of time from 1 to 3 years. P53 expression was detected in paraffin-embedded endometrial specimens by immunohistochemical methods. RESULTS: The percentage of endometrial hyperplasia positive for p53 expression was significantly greater in spontaneously occurring hyperplasia than in cases induced by the unopposed use of estradiol. CONCLUSION: Endometrial hyperplasia caused by the unopposed use of estradiol during menopause probably harbors fewer genomic errors than those cases occurring spontaneously.

c-erbB2 over-expression in endometrial hyperplasia induced by estrogens.

OBJECTIVE: To determine whether the presence of c-erbB2 expression in both spontaneous and estrogen-induced hyperplasia can affect the number of Ki-67-positive cells. PATIENTS AND METHODS: Thirty-two postmenopausal women with endometrial hyperplasia occurring spontaneously or after using unopposed estrogens were studied. The number of cells undergoing mitosis was estimated by immunohistochemical detection of the Ki-67 antigen and compared with the presence or absence of c-erbB2 over-expression. RESULTS: The percentage of cell nuclei showing positivity for Ki-67 was significantly higher in cases of endometrial hyperplasia that displayed c-erbB2 over-expression when compared to negative cases. CONCLUSION: The presence of c-erbB2 over-expression in endometrial hyperplasia is associated with a higher number of cells being positive for the Ki-67 proliferation marker.

Hysteroscopic and immunohistochemical findings in type I and type II endometrial carcinomas.

STUDY OBJECTIVE: To determine whether types I and II endometrial carcinomas have different precursor lesions recognized at hysteroscopy and immunohistochemistry. DESIGN: Single center case study (Canadian Task Force classification II-2). SETTING: A private, university affiliated hospital. PATIENTS: One hundred forty-six postmenopausal women with endometrial pathology diagnosed by hysteroscopy and biopsy. INTERVENTION: Hysteroscopy and immunohistochemical determination of p53 overexpression. MEASUREMENTS AND MAIN RESULTS: The number of cells showing p53 overexpression was low in endometrial polyps, hyperplasia, and G1 endometrioid carcinomas. A marked increase was seen only in high-grade G2-G3 endometrioid carcinomas and in cases of uterine papillary serous carcinoma. In the latter, strong p53 overexpression was detected in early forms still confined to endometrial polyps. Hysteroscopy could not differentiate between high- and low-grade endometrioid carcinomas. CONCLUSION: The p53-driven pathway plays an important role in the origin of papillary serous carcinoma but not in G1 endometrioid cancer that evolves from estrogen-stimulated endometrium. However, this pathway is important for progression of low-grade endometrioid carcinomas to higher-grade tumors.

[Differential expression of mucin carbohydrates in human endometria]. / Expresión diferencial de los carbohidratos de mucina en endometrios humanos.

The use of Lectins to identify oligosaccharides in mucin substances has been increased by the role played by cell surface carbohydrates in invasion and metastasis processes. We studied in this work normal endometrial tissue, with benign and malignant entities in search for the presence of the Galactose beta 1-3 N Acetylgalactosamine(Gal beta 1-3 GalNAC alpha and Galactose beta 1-3 N Acetylgalactosamine (Gal beta 1-3 alpha and beta) using the Lectins: Agaricus bisporus (ABL) and Arachis hipogea (PNA) respectively. The specific control were baths with galactose for PNA and with porcine stomach mucin for ABL. The use of these two Lectins allowed to differentiate substances bonded or non bonded to Sialic Acid, since PNA fails to label when the oligosaccharide is bonded to this acid Sialic. Significant differences were noticed on the bonding patterns of both Lectins on tissues with benign, malignant and normal entities. In this latter case the labelling was always continuous in both Lectins whereas it was irregular in the carcinoma.

Expresion diferencial de los carbohidratos de mucina en endometrios humanos / Differential expression of mucin carbohydrates in human endometriuns

La aplicación de Lectinas para la identificación de oligosacáridos constituyentes de las Mucinas presentes en la superficie celular, es una herramienta técnica muy útil, debido a que, probablemente estas sustancias estén involucrados en procesos tales como invasión y metástasis. En este trabajo, nosotros estudiamos tejido endometrial normal com entidades benignas y malignas, para investigar la presencia de Galactosa beta 1-3 N Acetilgalactosamina (Galbeta 1-3GalNacalpha) y de Galactosa beta 1-3 N Acetilgalactosamina(Galbeta 1-3GalNa alpha y beta) empleando dos Lectinas: Agaricus bisporus (ABL) y Arachis hipogea (PNA) respectivamente. Lo controles fueron realizados com baños de galactosa para PNA y com mucina porcina de estómago de cerdo para ABL. El uso de estas dos Lectinas, permitió encontrar diferencias en los patrones de unión, ya que si bien ambas se unen a los mismos oligosacáridos, ABL realiza la unión en presencia de Acido Siálico mientras que PNA no. Se observaron significativas diferencias en los patrones de unión de ambas lectinas en tejidos, com entidades benignas, malignas y tejido normal. En este último, la marcación fue simpre continua com ambas Lectinas, mientras que fue irregular en el carcinoma.

Expresion diferencial de los carbohidratos de mucina en endometrios humanos / Differential expression of mucin carbohydrates in human endometriuns

La aplicación de Lectinas para la identificación de oligosacáridos constituyentes de las Mucinas presentes en la superficie celular, es una herramienta técnica muy útil, debido a que, probablemente estas sustancias estén involucrados en procesos tales como invasión y metástasis. En este trabajo, nosotros estudiamos tejido endometrial normal com entidades benignas y malignas, para investigar la presencia de Galactosa beta 1-3 N Acetilgalactosamina (Galbeta 1-3GalNacalpha) y de Galactosa beta 1-3 N Acetilgalactosamina(Galbeta 1-3GalNa alpha y beta) empleando dos Lectinas: Agaricus bisporus (ABL) y Arachis hipogea (PNA) respectivamente. Lo controles fueron realizados com baños de galactosa para PNA y com mucina porcina de estómago de cerdo para ABL. El uso de estas dos Lectinas, permitió encontrar diferencias en los patrones de unión, ya que si bien ambas se unen a los mismos oligosacáridos, ABL realiza la unión en presencia de Acido Siálico mientras que PNA no. Se observaron significativas diferencias en los patrones de unión de ambas lectinas en tejidos, com entidades benignas, malignas y tejido normal. En este último, la marcación fue simpre continua com ambas Lectinas, mientras que fue irregular en el carcinoma. (AU)

Hormonal receptors in endometrial neoplasias.

The Authors studied the concentration of cytosolic and nuclear receptor for Estradiol and Progesterone in endometrial hyperplasia and carcinoma, compared with a control group. Comparative study of hormone receptor concentrations in different populations shows: 1) A significant increase in Estradiol receptors in endometrial hyperplasia (p less than 0.01); 2) A decreasing concentration of estradiol receptors with the decreasing of cellular differentiation in endometrial carcinoma (p less than 0.01); 3) A similar tendency and significance for Progesterone receptors; 4) For both receptors the tendency in the nuclear compartment is similar to that in cytosol but the significance is smaller (p less than 0.05).