Genistein effects on stromal cells determines epithelial proliferation in endometrial co-cultures

Background

Estrogen is the leading etiologic factor for endometrial cancer. Estrogen-induced proliferation of endometrial epithelial cells normally requires paracrine growth factors produced by stromal cells. Epidemiologic evidence indicates that dietary soy prevents endometrial cancer, and implicates the phytoestrogen genistein in this effect. However, results from previous studies are conflicting regarding the effects of genistein on hormone responsive cancers.

Methods

The effects of estrogen and genistein on proliferation of Ishikawa (IK) endometrial adenocarcinoma cells were examined in co-cultures of IK cells with endometrial stromal cells, recapitulating the heterotypic cell-to-cell interactions observed in vivo. The roles of estrogen receptor (ER)α and ERβ were evaluated using ERα and ERβ specific agonists. ER activation and cell proliferation in the IK epithelial cells were determined by alkaline phosphatase assay and Coulter counter enumeration, respectively.

Results

Both estrogen and genistein increased estrogen receptor-induced gene activity in IK cells over a range of concentrations. Estrogen alone but not genistein increased IK proliferation in co-cultures. When primed by estrogen treatment, increasing concentrations of genistein produced a biphasic effect on IK proliferation: nM concentrations inhibited estrogen-induced proliferation while μM concentrations increased proliferation. Studies with an ERβ-specific agonist produced similar results. Genistein did not influence the effects of estrogen on IK proliferation in monoculture.

Conclusions

Our study indicates that nutritionally relevant concentrations (nM) of genistein inhibit the proliferative effects of estrogen on endometrial adenocarcinoma cells presumably through activation of stromal cell ERβ. We believe that sub-micromolar concentrations of genistein may represent a novel adjuvant for endometrial cancer treatment and prevention.     

铜/低密度聚乙烯复合材料宫内节育器引起子宫异常出血与铜离子浓度和血管生成的相关性

BACKGROUND: To achieve excellent contraceptive effects, the copper wire on the copper-containing intrauterine device surface persistently releases copper ions, but ectopic intrauterine device, abnormal uterine bleeding, menostaxis and waist and abdominal pain are usually inevitable. OBJECTIVE: To investigate the correlation of abnormal uterine bleeding caused by copper/low-density polyethylene nanocomposite intrauterine device with copper ion concentration and expression of vascular endothelial growth factor in endometrial tissues. METHODS: Totally 60 female patients, aged from 25 to 40 years old, hospitalized for taking intrauterine devices were enrolled, and those patients were divided into abnormal uterine bleeding group (n=32) and non- bleeding group (n=28). In the meanwhile, another 20 women without intrauterine devices and with the normal menstrual cycle were selected as control group. Patients in the abnormal uterine bleeding group and non- bleeding group underwent the removal of intrauterine device and scraping endometrial tissues within 4-7 days after menstruation; patients in the control group underwent scraping endometrial tissues within 4-7 days after menstruation. The copper ion content in endometrial tissues was measured by atomic absorption method; expression of endometrial vascular endothelial growth factor detected by immunohistochemical staining and the microvessel density counted. Additionally, the corrosion ratio of copper ion in the simulated uterine fluid was detected using flame atomic absorption spectroscopy method. RESULTS AND CONCLUSION: The content and corrosion ratio of copper ion in the uterine abnormal bleeding group was significantly higher than those in the non-bleeding group (P < 0.05), and the copper ion content of no bleeding group was significantly higher than that of control group (P < 0.05). The amount of copper in the intrauterine device of abnormal uterine bleeding group was higher than that in non-bleeding group (P < 0.05). Compared with the control and non-bleeding groups, the vascular endothelial growth factor expression and microvessel density were significantly increased in the uterine abnormal bleeding group (P < 0.05); and the control group and non-bleeding group had no significant differences in the vascular endothelial growth factor expression and microvessel density. Furthermore, there was a positive correlation of the vascular endothelial growth factor expression with microvessel density and copper ion content. In conclusion, after implantation of copper/low-density polyethylene   nanocomposites intrauterine device, the higher copper ion concentration in endometrial tissues may lead to the over-expression of vascular endothelial growth factor that increases the endometrial microvessel density through a variety of ways, and promote microvessel expansion and congestion, finally resulting in abnormal uterine bleeding.     

Dual repressive effect of angiotensin II-type 1 receptor blocker telmisartan on angiotensin II-induced and estradiol-induced uterine leiomyoma cell proliferation

BACKGROUND: Although uterine leiomyomas or fibroids are the most common gynecological benign tumor and greatly affect reproductive health and well-being, the pathophysiology and epidemiology of uterine leiomyomas are poorly understood. Elevated blood pressure has an independent, positive association with risk for clinically detected uterine leiomyoma. Angiotensin II (Ang II) is a key biological peptide in the renin-angiotensin system that regulates blood pressure. METHODS: In this study, we investigated the potential role of Ang II (1-1000 nM) in the proliferation of rat ELT-3 leiomyoma cells in vitro. RT-PCR and western blot analysis with cell proliferation and DNA transfection assays were performed to determine the mechanism of action of Ang II. RESULTS: Ang II induced ELT-3 leiomyoma cell proliferation (P < 0.01) and the expression of Ang II type 1 receptor (AT(1)R) and AT(2)R mRNA and protein was confirmed. Regarding the intracellular signaling pathway, the Ang II-induced cell proliferation was AT(1)R-, epidermal growth factor receptor-, extracellular-regulated kinase- and protein kinase C-dependent but was not dependent on the AT(2)R or phosphatidylinositol-3 kinase or JAK kinase. The AT(1)R blocker telmisartan, effectively repressed Ang II-induced and estradiol-induced cell proliferation (P < 0.01). AT(1)R, but not AT(2)R, plays a role in Ang II-induced ELT-3 cell proliferation. CONCLUSIONS: These experimental findings in vitro highlight the potential role of Ang II in the proliferation of leiomyoma cells.     

Cyclin-dependent kinase inhibitor p27Kip1 controls growth and cell cycle progression in human uterine leiomyoma

The molecular mechanism of the cell-cycle machinery in uterine leiomyoma has not yet been fully elucidated. Among the various types of cell-cycle regulators, p27(Kip1) (p27) is considered to be a potent tumor suppressor. To provide further molecular basis for understanding the progression of uterine leiomyoma, our objective was to evaluate the expression level of p27 in normal myometrium and uterine leiomyoma tissue and its effect on cytogenic growth. Western blot analysis, real-time polymerase chain reaction (PCR) and immunohistochemical staining revealed that p27 protein and messenger RNA were down-regulated in uterine leiomyoma tissue and cultured cells compared to normal myometrium. Full-length human p27 cDNA was transferred using a replication-deficient recombinant adenoviral vector (Ad.p27) into uterine leiomyoma cells and evaluated the effect on cell proliferation. Transfection of Ad.p27 into uterine leiomyoma cells resulted in the induction of apoptosis, reduction in viability and proliferation of uterine leiomyoma cells. Our results suggest a new paradigm that down-regulated p27 protein expression is the possible underlying mechanism for the growth of uterine leiomyoma and over-expression of p27 induces cell death. This study provides better understanding of the control exerted by p27 in regulating growth and disease progression of uterine leiomyoma.     

Stimulatory and inhibitory effects of genistein on human uterine leiomyoma cell proliferation are influenced by the concentration

BACKGROUND: Due to dietary exposure of women to genistein, a soy-derived phytoestrogen, and the estrogen responsiveness of uterine leiomyomas 'fibroids', we evaluated the effects of genistein (0.001-50 microg/ml) on human uterine leiomyoma (UtLM) cells versus uterine smooth muscle cells (UtSMCs) in vitro. METHODS: Light microscopy was used to determine the effects of genistein on cell morphology. Proliferation was assessed using a colorimetric assay and proliferating cell nuclear antigen (PCNA) immunocytochemistry. Flow cytometry was used to quantitate cells in the S-phase and those undergoing apoptosis. A fluorometric assay and confocal microscopy were used to detect caspase-3 activity and apoptotic bodies, respectively. RESULTS: In UtLM cells, low concentrations (< or = 1 microg/ml) of genistein stimulated proliferation, increased PCNA labeling and the percentage of cells in the S-phase, but this did not occur in UtSMCs. Higher concentrations (> or = 10 microg/ml) of genistein adversely affected the morphology, significantly inhibited proliferation, decreased PCNA labeling, increased caspase-3 activity and induced apoptosis in both cell types. CONCLUSIONS: Genistein's effects are concentration-dependent in both cell lines. Lower concentrations elicit proliferative effects on UtLM cells only; whereas, higher concentrations alter morphology, inhibit proliferation, and increase caspase activity and apoptosis in both cell types, with the latter two effects being more extensive in UtSMCs.     

Contraception: Endometrial lymphomyeloid cells in abnormal uterine bleeding due to levonorgestrel (Norplant)

Endometrial lymphomyeloid cell subsets were evaluated in samplesfrom normal women and from women with abnormal uterine bleedingdue to subcutaneous levonorgestrel implants (Norplant) or anintrauterine device (IUD). The frequency of CD3⁺, CD68⁺, CD43⁺and endometrial granulated lymphold cells was evaluated by inimunohistochemicalor phloxine-tartrazine staining of formalinfixed paraffin-embeddedsamples. In normal women, cyclic variation in lymphomyeloidsubsets was seen. In women using Norplant for contraception,the frequency of CD3⁺, CD68⁺ and CD43⁺ cells was dramaticallydecreased, compatible with endometrial atrophy. When Norplantusers with abnormal bleeding were compared to women withoutbleeding, however, the number of CD68⁺ cells was significantlyincreased and the number of CD3⁺ and CD43⁺ cells was preserved,contrary to the hypothesis that this group would show a greaterdegree of atrophy and hence, tissue fragillty. A similar patternwas seen in a preliminary study of women with bleeding associatedwith use of copper-only IUD contraception, and in samples takenfrom late secretory and menstrual phase biopsies from normalcycling women. Whether these changes in endometrial lymphomyeloidcells represent a result of bleeding arising from a common mechanismor rather cause the uterine bleeding is discussed.     

Clinical significance of histiocytes in the detection of endometrial adenocarcinoma and hyperplasia

The purpose of this case-control study was to determine the clinical significance of histiocytes and normal endometrial cells as cytologic markers of adenocarcinoma or hyperplasia of the endometrium. Cervical-vaginal smears obtained in 102 patients with mean age 59.7 yr with abnormal uterine bleeding and endometrial pathology, and 101 controls with mean age 56.5 yr with postmenopausal bleeding and whose cytologic smear was negative, were evaluated. Histiocytes alone failed to predict either endometrial adenocarcinoma (odds ratio [OR], 1.02; 95% confidence interval [CI], 0.32–3.22) or hyperplasia (OR, 1.10; 95% CI, 0.37–3.30). The odds of endometrial carcinoma in symptomatic, postmenopausal women was three times greater in the presence of histiocytes with phagocytosis of acute inflammatory cells (PIC) (OR, 3.00; 95% CI, 1.16–7.70). Endometrial hyperplasia was more frequently observed when cervical smears contained normal endometrial cells (OR, 4.09; 95% CI, 1.14–14.67). Only histiocytes with PIC and normal endometrial cells carry a three- and fourfold greater likelihood of coexistent endometrial adenocarcinoma and hyperplasia, respectively. Such strong association may require endometrial biopsy or transvaginal ultrasonography to uncover lesional tissue in the endometrial cavity. Diagn. Cytopathol. 1998;19:89–93. © 1998 Wiley-Liss, Inc.     

The effects of the selective progesterone receptor modulator asoprisnil on the morphology of uterine tissues after 3 months treatment in patients with symptomatic uterine leiomyomata

BACKGROUND: Asoprisnil is a selective progesterone receptor modulator with mixed progesterone agonist/antagonist activity which controls uterine bleeding via an endometrial effect. This study examined full-thickness endometrial, leiomyoma and myometrial morphology in hysterectomy specimens from patients with uterine leiomyomata, after treatment with asoprisnil for 3 months. METHODS: In this double-blind, randomized, placebo-controlled study, 33 subjects with uterine leiomyomata were randomized to receive asoprisnil 10, 25 mg or placebo for an average of 95 days prior to hysterectomy. Samples of endometrium, myometrium and leiomyoma tissue were subjected to systematic morphological assessment with quantification of mitotic activity. RESULTS: In patients treated with 10 or 25 mg asoprisnil, a unique pattern called 'non-physiologic secretory effect' was evident in endometrium, recognizable through partially developed secretory glandular appearances and stromal changes. Endometrial thickness was decreased, and there were low levels of mitotic activity in endometrial glands and stroma. Unusual thick-walled muscular arterioles and prominent aggregations of thin-walled vessels were present in endometrial stroma, but not in myometrium or non-endometrial vascular beds. Mitotic activity was decreased in leiomyomata. CONCLUSIONS: Asoprisnil induces unique morphological changes and is associated with low levels of glandular and stromal proliferation in endometrium, and in leiomyomata. These changes are likely to contribute to the amenorrhoea experienced after exposure to the medication.     

The Role of Trophoblast Interferons in the Maintenance of Early Pregnancy in Ruminants

PROBLEM: Are the effects of ruminant trophoblast interferon-tau (IFN-τ) on uterine prostaglandin (PG) secretion a specific action of this cytokine and what are the effects of IFN-τ on expression of uterine genes not generally associated with pregnancy maintenance? METHODS: The effects of IFN-τ and IFN-α on bovine uterine expiant and epithelial cell production of PGF_2α and PGE₂ were determined in the presence and absence of oxytocin (OT). The effects of intrauterine administration of IFN-τ were determined on uterine expression of retinol-binding protein (RBP) and transforming growth factor-beta (TGF-β) isoforms. RESULTS: IFN-τ attenuated uterine endometrial secretion of PGF_2α and PGE₂ in vitro and diminish PG stimulation by OT. IFN-τ and IFN-α were observed to be equipotent. Intrauterine infusion of IFN-τ resulted in a significant decrease in steady-state RBP mRNA levels and expression of TGF-B1, 2, and 3 mRNA levels were lowest in IFN-τ treated animals. CONCLUSION: Negative regulation of gene expression may be a general strategy in IFN activity. This may explain the similar activities of IFN-τ and IFN-α on a broad variety of cell types, including ruminant uterine endometrium.     

Neural tube defects and impaired neural progenitor cell proliferation in Gβ1‐deficient mice

Heterotrimeric G proteins are well known for their roles in signal transduction downstream of G protein–coupled receptors (GPCRs), and both Gα subunits and tightly associated Gβγ subunits regulate downstream effector molecules. Compared to Gα subunits, the physiological roles of individual Gβ and Gγ subunits are poorly understood. In this study, we generated mice deficient in the Gβ1 gene and found that Gβ1 is required for neural tube closure, neural progenitor cell proliferation, and neonatal development. About 40% Gβ1^?/? embryos developed neural tube defects (NTDs) and abnormal actin organization was observed in the basal side of neuroepithelium. In addition, Gβ1^?/? embryos without NTDs showed microencephaly and died within 2 days after birth. GPCR agonist‐induced ERK phosphorylation, cell proliferation, and cell spreading, which were all found to be regulated by Gαi and Gβγ signaling, were abnormal in Gβ1^?/? neural progenitor cells. These data indicate that Gβ1 is required for normal embryonic neurogenesis. Developmental Dynamics 239:1089–1101, 2010. © 2010 Wiley‐Liss, Inc.     

The influence of extracellular matrix proteins on T-cell proliferation and apoptosis in women with endometriosis or uterine leiomyoma

PROBLEM: Interactions between the extracellular matrix (ECM) and peripheral blood T cells in women with endometriosis and leiomyoma are hardly unknown. We have investigated the influence of two major ECM components, collagen IV (C-IV) and fibronectin (Fn), on T-cell proliferation and apoptosis in women with endometriosis and uterine leiomyoma. beta1 integrin expression, responsible for interactions with ECM proteins, was also studied. METHOD OF STUDY: Peripheral blood lymphocytes were obtained from 53 women (17 with uterine leiomyomas, 18 with endometriosis, and 18 from healthy donors). T cells were exposed to ECM proteins co-immobilized with monoclonal antibody anti-CD3 for 72 hr. Apoptosis and S phase of the cell cycle of the T cells were studied by DNA analysis using flow cytometry. The proliferation of T cells was evaluated by MTT assay. The percentage of CD3+ cells expressing CD29 (beta1 integrin chain) was evaluated by double-color flow cytometry. Results were analyzed statistically using the Mann-Whitney test. RESULTS AND CONCLUSIONS: (1) A general increase in the percentage of T cells in S phase could be seen in women with endometriosis and uterine leiomyoma in all culture conditions what may suggest general activation of T cells. (2) A significant increase in the percentage of cells in S phase was shown only in the case of T cells exposed to anti-CD3 + C-IV in both women with uterine leiomyoma and endometriosis. (3) However, no apoptotic cells were observed. (4) T cells from patients with uterine leiomyoma exhibited significantly increased level of proliferation after culture with anti-CD3 + C-IV. (5) More T cells expressed beta1 integrin in women with endometriosis or uterine leiomyoma than in healthy donors. Our data may suggest that increased beta1 integrin expression may enhance T-cell-ECM interactions, which may be responsible for the increased proliferation of T cells but not for apoptosis. Therefore, it is possible that interactions of T cells with ECM proteins, especially with C-IV, may contribute to the pathogenesis of endometriosis and uterine leiomyoma.     

Increased Expression of Cell Surface Markers on Endometrial γδ T-Cell Receptor+ Intraepithelial Lymphocytes Induced by the Local Presence of the Sheep Conceptus

PROBLEM: In sheep, γδ T-cell receptor (TCR)⁺ cells are a major lymphocyte subpopulation in the luminal epithelium of the uterine endometrium. During late pregnancy, this population of T cells increases in number and becomes more granulated. This study was performed to determine whether this apparent activation was induced by local effects of the conceptus or systemic effects of pregnancy. METHODS: The unilaterally-pregnant ewe, in which the conceptus is surgically confined to one uterine horn, was used to distinguish between systemic and local effects of pregnancy on function of endometrial γδ TCR⁺ intraepithelial lymphocytes. Lymphocytes collected from peripheral blood, and from the endometrial luminal epithelium of cyclic and unilaterally-pregnant ewes (day 140 of gestation) were analyzed by flow cytometry. RESULTS: As compared to γδ TCR⁺ lymphocytes in peripheral blood, γδ TCR⁺ intraepithelial lymphocytes from non-pregnant ewes had a lower percentage of cells staining positive for CD25, CD44, and L-selectin. There was no effect of pregnancy on the percentage of γδ TCR⁺ peripheral blood lymphocytes staining positive for CD25, CD44, CD29, or L-selectin. Similarly, the percentage of intraepithelial lymphocytes staining positive for these antigens was similar for cells collected from cyclic ewes and cells from the nonpregnant uterine horn of unilaterally-pregnant ewes. In contrast, there was a higher percentage of CD25, CD44, CD29, and L-selectin⁺ cells for γδ TCR⁺ intraepithelial lymphocytes from the conceptus-containing uterine horn of pregnant ewes than for γδ TCR⁺ intraepithelial lymphocytes from the endometrium of cyclic ewes or from the nonpregnant uterine horn of pregnant ewes. CONCLUSION: The local presence of the conceptus causes changes in cell surface marker expression on endometrial γδ TCR⁺ intraepithelial lymphocytes during pregnancy. These changes may reflect conceptus-induced activation of this population of cells.     

A novel selective progesterone receptor modulator asoprisnil (J867) down-regulates the expression of EGF, IGF-I, TGFbeta3 and their receptors in cultured uterine leiomyoma cells

BACKGROUND: This study was conducted to evaluate the effects of a novel selective progesterone receptor modulator (SPRM) asoprisnil on the expression of growth factors and their receptors and on growth factor-induced proliferation of cultured uterine leiomyoma and matching myometrial cells. METHODS: The expression of epidermal growth factor (EGF), insulin-like growth factor-I (IGF-I) and transforming growth factor (TGFbeta3) was assessed by immunocytochemistry and semi-quantitative RT-PCR. The expression of phosphorylated EGF receptor (p-EGFR), IGF-I receptor alpha subunit (IGF-IRalpha) and phosphorylated TGFbeta receptor type II (p-TGFbeta RII) was assessed by Western blot analysis. Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay. RESULTS: Treatment with 10(-7) M asoprisnil decreased EGF, IGF-I and TGFbeta3 mRNA and protein expression as well as p-EGFR, IGF-IRalpha and p-TGFbeta RII protein expression in leiomyoma cells cultured for 72 h. EGF (100 ng/ml), IGF-I (100 ng/ml) and TGFbeta3 (10 ng/ml) increased the number of viable leiomyoma cells cultured for 72 h, whereas the concomitant treatment with 10(-7) M asoprisnil antagonized the growth factor-induced increase in leiomyoma cell proliferation. In cultured myometrial cells, however, asoprisnil affected neither the growth factor and their receptor expression nor the cell proliferation. CONCLUSION: Asoprisnil inhibits the expression of EGF, IGF-I, TGFbeta3 and their receptors in cultured leiomyoma cells without affecting their expressions in myometrial cells.     

Keratinocyte Growth Factor Stimulates Macrophage Inflammatory Protein 3α and Keratinocyte‐derived Chemokine Secretion by Mouse Uterine Epithelial Cells

Citation Haddad SN, Wira CR. Keratinocyte growth factor stimulates macrophage inflammatory protein 3α and keratinocyte‐derived chemokine secretion by mouse uterine epithelial cells. Am J Reprod Immunol 2010; 64: 197–211 Problem Communication between uterine epithelial cells and the underlying stromal fibroblasts is critical for proper endometrial function. Stromal fibroblast‐derived growth factors have been shown to regulate epithelial immune functions. The purpose of this study was to determine whether keratinocyte growth factor (KGF) regulates uterine epithelial cell chemokine and antimicrobial secretion. Method of study Uterine epithelial cells were isolated from Balb/c mice and cultured in either 96‐well plates or transwell inserts. Epithelial cells were treated with KGF, epidermal growth factor (EGF), or hepatocyte growth factor (HGF). Macrophage inflammatory protein 3α (MIP3α) and keratinocyte‐derived chemokine (KC) levels were measured by ELISA. Results Keratinocyte growth factor stimulated the secretion of MIP3α and KC. The effects on MIP3α by KGF were specific because EGF and HGF had no effect. In contrast, KGF, EGF, and HGF had similar effects on KC. Furthermore, KGF administered to the apical side of epithelial cells had no effect on MIP3α or KC secretion, indicating that the KGF receptor is located on the basolateral surface of uterine epithelial cells. Conclusion We demonstrate that KGF plays a role in uterine epithelial cell secretion of MIP3α and KC, key immune mediators involved in the protection of mucosal surfaces in the female reproductive tract.     

Effect of human endometrial stromal cell-derived conditioned medium on uterine natural killer (uNK) cells' proliferation and cytotoxicity

Citation Chen Y, Zhuang Y, Chen X, Huang L. Effect of human endometrial stromal cell‐derived conditioned medium on uterine natural killer (uNK) cells’ proliferation and cytotoxicity. Am J Reprod Immunol 2011; 65: 589–596 Problem Human endometrial stromal cells are involved in the regulation of immune cell proliferation, apoptosis, differentiation, and function. In the endometrium, uNK cells are in close contact with stromal cells. The aim of the study was to investigate the effects of human endometrial stromal cells on uNK‐cell proliferation and uNK‐cell cytotoxicity. Method of study The conditioned medium was derived from the endometrial stromal cells in the proliferative phase, secretory phase, and early pregnancy. The effects of stromal cell‐derived conditioned medium on uNK‐cell proliferation and cytotoxicity were detected by mitochondrial lactate dehydrogenase‐based MTS staining and flow cytometry. Results The stromal cell‐derived conditioned medium in both secretory phase and early pregnancy significantly promoted uNK‐cell proliferation. Compared with the control group, the uNK‐cell cytotoxicity were significantly reduced by conditioned medium in the proliferative, secretory, and decidua groups, but there were no significant differences among these different physiological stages in the inhibiting ability. Conclusion Human endometrial stromal cells may be involved in the regulation of uNK‐cell functions through influencing proliferation and cytolytic activity.     

Hysteroscopy for asymptomatic postmenopausal women with sonographically thickened endometrium

Endometrial carcinoma is the most common genital cancer in women. While patients usually present with vaginal bleeding, in 10–20% this characteristic symptom is absent. Endometrial thickness (double layer) is measured by transvaginal sonography and thickening indicates an increased risk of malignancy or other pathology (hyperplasia or polyps). Objective: We sought to correlate hysteroscopic and pathological findings in asymptomatic postmenopausal women with sonographically thickened endometrium (>6 mm). Study design: A prospective observational study in a university hospital of 304 postmenopausal women referred between 1996 and 2006 because of a sonographically thickened endometrium in the absence of abnormal bleeding, who underwent continuous flow hysteroscopy (4.5 mm Storz hysteroscope) and fractionated curettage of the uterine cervix and corpus (D & C) in addition to vaginal sonography (5 MHz probe). Results: The mean age of the women was 64.8 (range 57.7–71.9) years. Average endometrial thickness measured by ultrasound was 12 mm ± 6.7 mm. Hysteroscopy suggested the presence of endometrial polyps in 226 women (74.3%), simple endometrial hyperplasia in 34 (11.2%), atrophic endometrium in 18 (5.9%), complex endometrial hyperplasia in 2 (0.7%), atypical hyperplasia in 3 (1%) and leiomyoma in 9 (3.0%). In 12 women (3.9%), the hysteroscopic appearance suggested malignancy and histology revealed endometrial adenocarcinoma. All hysteroscopic results were confirmed by histological examination. Conclusion: Hysteroscopy represents an easy, safe and effective method for the investigation of asymptomatic women with a thickened endometrium found with transvaginal ultrasound. The commonest pathology was endometrial polyps.     

Endometrial assessment in women using tibolone or placebo: 1-year randomized trial and 2-year observational study

OBJECTIVE: To assess the effects of tibolone on the endometrium of postmenopausal women. DESIGN: A 1-year randomized, double-blind, placebo-controlled clinical trial and a 2-year open clinical trial. The placebo-controlled trial included 40 participants: 20 in the placebo group and 20 in the tibolone group; in the open trial, 17 participants receiving tibolone were assessed over 24 months. Transvaginal ultrasonography was carried out to assess endometrial thickness, and endometrial appearance was assessed on hysteroscopy. In addition, endometrial samples were submitted to histological examination. The occurrence of uterine bleeding and other adverse effects was also assessed. RESULTS: Results suggest that tibolone does not exert a stimulatory effect on the endometrium: unaltered endometrial thickness, atrophic appearance of most endometria on hysteroscopy, and endometrial histology classified as atrophic, hypotrophic with incipient secretion, or hypotrophic with weak proliferation (one case). Tibolone was effective in the treatment of climacteric symptoms, and only 8.7% of the participants presented uterine bleeding during treatment. CONCLUSIONS: Tibolone seems to be an effective option for the treatment of climacteric symptoms in postmenopausal women, especially in women who do not want to experience uterine bleeding again.     

Transvaginal Doppler sonography: is there a role for this modality in the evaluation of women with postmenopausal bleeding?

OBJECTIVE: The aim of this study was to determine whether transvaginal sonography (TVS) and transvaginal Doppler sonography (TDS) can discriminate between normal and abnormal endometrium. MATERIALS AND METHODS: Patients who had vaginal bleeding an year after menopause and were not on HRT or tamoxifen were preoperatively examined by TVS and TDS on the same day of curettage. The endometrial thickness as well as the pulsatility index (PI) and resistance index (RI) of uterine arteries were recorded. RESULTS: Final pathology analysis revealed that 55/81 (67.9%) had normal endometrial tissue and 26/81 (32.1%) had an abnormality (endometrial hyperplasia, polyp or cancer). The mean endometrial thickness was greater in the abnormal group (9.4 mm versus 3.8 mm, P < 0.05). The mean PI of normal and abnormal endometrium were 2.85 and 1.53. The mean RI of normal and abnormal endometrium were 1.04 and 0.68. Univariate analysis found that PI < 2 and RI < 0.9 were correlated with abnormal endometrium (P < 0.05). Multivariate analysis revealed significance only for the endometrial thickness. CONCLUSIONS: TDS cannot distinguish between normal and abnormal endometrium. Using TVS only would result in a significant reduction of endometrial biopsy or curettage.     

Plasma cells in chronic endometritis are easily identified when stained with syndecan-1.

BACKGROUND: Chronic endometritis has been observed in 3-10% of women with irregular uterine bleeding who undergo endometrial biopsy. The diagnosis of chronic endometritis rests on the recognition of plasma cells in endometrial tissue that may show a prominent spindle cell stromal component, and is frequently difficult to date. Syndecan-1 is a cell-surface proteoglycan that is expressed on the cell surface of plasma cells. DESIGN: Eighteen endometrial curettage cases with the diagnosis of chronic endometritis and 25 endometrial curettage cases of dysfunctional uterine bleeding, in females under the age of thirty-five in whom no other histopathologic changes were noted, were reviewed for the presence of plasma cells. Sections were then stained with syndecan-1. RESULTS: All of the chronic endometritis cases showed easily visible syndecan-1 staining of plasma cell membranes. None of the cases of dysfunctional uterine bleeding showed presence of plasma cells in either the hematoxylin and eosin stained or syndecan-1 stained sections. CONCLUSIONS: In cases of suspected chronic endometritis in which no plasma cells can be found on hematoxylin and eosin stained slides, syndecan-1 may be an effective adjunct in the identification of plasma cells and thus aid in the diagnosis of chronic endometritis.     

Decreased tissue inhibitor of metalloproteinase in the endometrium of women using depot medroxyprogesterone acetate: a role for altered endometrial matrix metalloproteinase/tissue inhibitor of metalloproteinase balance in the pathogenesis of abnormal uterine bleeding?

BACKGROUND: Abnormal uterine bleeding is commonly associated with progestin-only contraceptives, including depot medroxyprogesterone acetate (DMPA), and remains the main reason why these agents are discontinued. Matrix metalloproteinases (MMP), enzymes which degrade specific extracellular matrix components, and leukocytes are implicated in menstruation. Alteration in endometrial MMP-9 and leukocytes has been described in users of other progestin-only contraceptives, suggesting a potential role in the pathogenesis of abnormal uterine bleeding. METHODS: This study describes the immunohistochemical localization of MMP-9, the tissue inhibitors of metalloproteinases (TIMP)-1, TIMP-2 and TIMP-3, and leukocytes [CD3+ T lymphocytes, CD68+ macrophages and CD56+ uterine natural killer cells (uNK cells)] in the endometrium of women using DMPA. Comparison is made with perimenstrual endometria from normal cycling women. RESULTS: Similar to the perimenstrual period, an influx of MMP-9 positive cells (identified as neutrophils and CD3+ T cells on the basis of dual immunofluorescence), macrophages and uNK cells was observed in the endometrium of DMPA users. However, significantly more endometrial T lymphocytes were observed in DMPA users. Immunoreactive TIMP, present in all endometrial compartments, demonstrated a significantly decreased immunostaining intensity score in endometrial epithelium (TIMP-1 and TIMP-2), stroma (TIMP-1, TIMP-2 and TIMP-3), endothelium (TIMP-1 and TIMP-2) and vascular smooth muscle (TIMP-1) of DMPA users compared with controls. No correlation was observed between the parameters studied and bleeding patterns reported by subjects. CONCLUSIONS: These findings provide additional evidence for the importance of the MMP/TIMP balance in the loss/maintenance of endometrial integrity and in the complex pathological mechanisms involved in the troubling side-effect of menstrual bleeding disturbance.