Journal of the Society for Gynecologic Investigation最新文献

Objective: The current study sought to compare the endometrial localization of the integrin subunit alpha-6 in women with endometriosis and women without the disease. Alpha-6 integrins have an important function, not only in the attachment of cells to the extracellular matrix and laminin, but they also serve as inductors of cell migration and invasion, depending on their pattern of expression in the cell membrane.

Methods: The endometriosis group consisted of 32 women with a confirmed diagnosis of endometriosis by laparoscopy or laparotomy. The control group consisted of 20 women not having endometriosis or any other gynecologic disease at laparoscopy. Endometria were obtained by biopsy. Immunohistochemical techniques were used to assess alpha-6 localization. In each section, the percentage of positive cells and the localization of expression were evaluated.

Results: All glandular cells expressed alpha-6 in all of the samples but presented two different patterns, either only in the basal side of the cells (polarized) or also in other sides of the cells (depolarized). The percentage of samples showing depolarized expression was significantly higher in the endometriosis group (66.6% vs 15.8%, chi2 =12.09, P = .001).

Conclusions: The endometria of women with endometriosis more frequently show a depolarized expression of integrin subunit alpha-6, a characteristic usually found in highly proliferating cells with migrating and invasive abilities.

Objective: To evaluate the clinical features and the expression of transforming growth factor-beta3 (TGF-beta3) and connective tissue growth factor (CTGF) in myometrium and uterine leiomyomas after preoperative treatment with gonadotropin-releasing hormone-analogs (GnRH-a) and tibolone.

Methods: Twenty-three patients received 3.75 mg leuprolide acetate depot for 4 months. Twenty-two patients received the same therapy plus 2.5 mg tibolone daily. Patients underwent uterine surgery after therapy. Twenty-two untreated patients underwent surgery directly. Hematologic tests, bone mineral density (BMD) measurement, and ultrasonographic evaluation of uterine volume were performed before and after treatment. Menorrhagia and pelvic pain were evaluated with a visual analog scale. Hot flushes were recorded in daily diaries. Immunohistochemical expression of TGF-beta3 and CTGF in myometrium and myoma samples was evaluated semiquantitatively.

Results: After therapy, hemoglobin and iron levels similarly increased in both groups. BMD significantly decreased only in the GnRH-a group. Uterine volume similarly decreased in both groups. No patient had menorrhagia or pelvic pain at the end of therapy. The number of hot flushes increased after the first month in the GnRH-a group; in the GnRH-a plus tibolone group, it remained constant and was lower. In untreated cases, TGF-beta3 and CTGF smooth muscle cell immunoexpression was lower in myometrium than in leiomyomas. After medical treatment, growth factor immunoexpression remained unchanged in myometrial samples and was reduced in leiomyomas. Endothelial cells showed strong immunopositivity, both in untreated and in treated cases.

Conclusion: This study focuses on the effects of GnRH-a and tibolone on TGF-beta3 and CTGF expression in myometrium and myomas and supports the hypothesis of a pathogenetic role of these growth factors in uterine fibromatosis.

Objectives: The majority of endometrial cancers arise as a result of estrogen stimulation, the molecular targets of which remain incompletely defined. We hypothesize that the granulin-epithelin precursor (GEP) may be one such target. In this study, we examined the frequency of GEP and estrogen receptor (ER) co-expression in human endometrial cancers. Once we established the co-expression of GEP with the estrogen receptor we examined the potential estrogen regulation of GEP expression, as well as the functional significance of GEP expression in vitro.

Methods: Double immunofluorescence and confocal microscopy were used to compare GEP and ER expression among 41 endometrial cancers. The effects of estradiol and tamoxifen treatment on GEP expression in two endometrial cancer cell lines, KLE and HEC-1-A, were assessed through reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis. The antiproliferative effect of GEP silencing by short hairpin (sh)RNA, was evaluated in HEC-1-A cells using an MTT assay.

Results: GEP co-expression with ER was observed in 63% of the cancers examined. A two- to fivefold increase in GEP expression with estradiol and/or tamoxifen treatment was observed in KLE cells. Silencing of GEP in HEC-1-A cells using shRNA resulted in a decrease in proliferation among transfected cells.

Conclusions: Co-expression of GEP and ER in endometrial cancer cells, and the regulation of GEP by estrogen, suggests a role for GEP in steroid-mediated endometrial cancer cell growth. Further characterization of GEP as a steroid-mediated growth factor in these cells may broaden our understanding of endometrial cancer biology and also provide guidance in the development of novel therapeutic targets.

Objective: Successful placental development is crucial for optimal growth, maturation, and survival of the embryo/fetus. To examine genetic aspects of placental development, we investigated gene expression patterns in the murine placenta at embryonic day 10.5 (E10.5), E12.5, E15.5, and E17.5.

Methods: By use of the Affymetrix MU74A array (Affymetrix, Santa Clara, CA), we measured expression levels for 12,473 probe sets. Using pairwise analysis we selected 622 probe sets, corresponding to 599 genes, that were up- or down-regulated by more than fourfold between time points E10.5 and E12.5, E12.5 and E15.5, E15.5 and E17.5. We analyzed and functionally annotated those genes regulated during development.

Results: In comparing E10.5 to E12.5 we found that angiogenesis and fatty acid metabolism and transport related genes were up-regulated at E10.5, while genes involved in hormonal control and ribosomal proteins were up-regulated at E12.5. When comparing E12.5 to E15.5 we noted that genes involved in the cell cycle and RNA metabolism were strongly up-regulated at E12.5, while genes involved in cellular transport were up-regulated at E15.5. Finally, when comparing E15.5 to E17.5, we found genes related to cell cycle control, genes expressed in the nucleus and involved in RNA metabolism were up-regulated at E17.5.

Conclusion: Microarray analysis has allowed us to describe gene expression patterns and profiles in the developing mouse placenta. Further analysis has demonstrated that several functional classes are up- and down-regulated at specific time points in placental development. These changes may have significant implications for placental development in the human.

Objective: Pentraxin-3 (PTX3) is a long pentraxin that plays a key role in female fertility as a structural and essential constituent of the cumulus oophorus extracellular matrix. Despite considerable evidence supporting this role of PTX3 in mice, data in humans are scanty. The aim of the present study was (1) to evaluate follicular fluid concentrations of PTX3; (2) to test the hypothesis that levels of the molecule correlate with oocyte characteristics (corona radiata, aspect of the cumulus, nuclear maturity, and fertilization); and (3) to evaluate the possibility that peripheral concentration of PTX3 may be of clinical help in monitoring ovarian hyperstimulation.

Methods: ELISA was used to determine PTX3 concentration. Levels of PTX3 were tested in 96 follicles.

Results: The mean +/- SD and the median (interquartile range) were 17.9 +/- 18.3 and 12.1 (6.5-23.6) ng/mL, respectively. Levels of the molecule did not appear to be normally distributed. At the day of ovum pick-up, levels of PTX3 were 6.3-fold higher in follicular fluid than in peripheral blood (95% CI, 3.6-9.0). No statistically significant difference emerged linking follicular fluid concentration of PTX3 and oocyte quality. In a series of ten women, plasma concentration of PTX3 did not vary during ovarian hyperstimulation, resulting in levels of 1.0 +/- 0.5 at the 3rd day of the menstrual cycle and 1.0 +/- 0.6 ng/mL at the day of oocyte retrieval.

Conclusions: Results from the present study support the following conclusions: (1) elevated levels of soluble PTX3 can be found in follicular fluid; (2) follicular fluid concentration of PTX3 cannot by used as a marker of oocyte quality; and (3) plasma concentration of the molecule is not influenced by ovarian hyperstimulation.

Objectives: The controversy regarding potential long-term side effects of antenatal steroid administration for accelerating fetal lung maturation is still unresolved despite more than 30 years of experience. Studies in animals have demonstrated that administration of glucocorticoids during pregnancy alters renal expression of several key regulatory molecules at different developmental stages followed in most cases with the development of hypertension in the adult. We studied the effects of betamethasone on the expression of (1) NA,K-ATPAse pump; (2) the Na/H exchanger 3 (NAHE3); (3) angiotensin receptor (AT1 and AT2); and (4) the type 1 dopamine receptor (D1R).

Methods: Pregnant sheep were treated with either 0.17 mg/kg betamethasone or vehicle 24 hours apart at 80 and 81 days' gestation. Fetal kidneys were harvested at 81 and 135 days' gestation. Protein and mRNA levels were measured in kidney cortex.

Results: Betamethasone had acute and long-term effects on fetal kidney cortex gene expression. Acutely, mRNA abundance for AT2 was significantly lower and that of NHE3 significantly higher than in controls (0.4 +/- 0.02 vs 0.7 +/- 0.05; 1.2 +/- 0.16 vs 0.4 +/- 0.04; P < .05). At 135 days' gestation, AT2 receptor abundance remained lower than control (0.2 +/- 0.02 vs 0.4 +/- 0.02; P < .05), whereas D1R expression was higher (0.8 +/- 0.17 vs 0.5 +/- 0.06; P < .05). No changes in Na,K-ATPase of AT1 receptor at either of the two time points studied were observed. Antenatal steroid administration was not associated with premature labor or a reduction in either body weight or kidney weight.

Conclusion: Our findings strongly suggest that antenatal glucocorticoid administration according to National Institutes of Health (NIH) consensus guidelines may alter human fetal renal development. Further studies are needed to establish a direct relationship between alterations in fetal renal gene expression and the development of hypertension in adulthood.

Objective: Nuchal edema (NE) is a clinical indicator for aneuploidy, cardiovascular anomalies, and several genetic syndromes. Its etiology, however, is unknown. In the nuchal area, the endothelium of the jugular lymphatic sacs (JLS) develops by budding from the blood vascular endothelium of the cardinal veins. Abnormal distension of the jugular sacs is associated with NE. We hypothesize that a disturbed lymphatic endothelial differentiation and sac formation causes NE. We investigated endothelial differentiation of the jugular lymphatic system in human and mouse species with NE.

Methods: Aneuploid human fetuses (trisomy 21; trisomy 18) were compared with euploid controls (gestational age 12 to 18 weeks). Trisomy 16 mouse embryos were compared with wild type controls (embryonic day 10 to 18). Trisomy 16 mice are considered an animal model for human trisomy 21. Endothelial differentiation was investigated by immunohistochemistry using lymphatic markers (prox-1, podoplanin, lymphatic vessel endothelial hyaluronan receptor [LYVE]-1) and en blood vessel markers (neuropilin [NP]-1 and ligand vascular endothelial growth factor [VEGF]-A). Smooth muscle actin (SMA) was included as a smooth muscle cell marker.

Results: We report a disturbed venous-lymphatic phenotype in aneuploid human fetuses and mouse embryos with enlarged jugular sacs and NE. Our results show absent or diminished expression of the lymphatic markers Prox-1 and podoplanin in the enlarged jugular sac, while LYVE-1 expression was normal. Additionally, the enlarged JLS showed blood vessel characteristics, including increased NP-1 and VEGF-A expression. The lumen contained blood cells and smooth muscle cells lined the wall.

Conclusion: A loss of lymphatic identity seems to be the underlying cause for clinical NE. Also, abnormal endothelial differentiation provides a link to the cardiovascular anomalies associated with NE.

Objectives: The purpose is to investigate how umbilical arterial blood flow changes by an intraamniotic distilled water infusion and to determine whether the changes in umbilical circulation have any relationship with fetal cardiovascular status and osmolality in amniotic fluid and fetal plasma.

Methods: Eleven chronically catheterized pregnant sheep were used in this study. After a 1-hour control period, 1.5 L of warmed sterile distilled water was injected over 10 minutes into the amniotic cavity. Fetal heart rate and carotid arterial pressure, blood flow of the umbilical and fetal carotid arteries were continuously measured. Fetal arterial blood sampled twice during the control period and then at 30, 60, 90, 120, 180, 240, 300, and 360 minutes after the start of the infusion, was analyzed for blood gases, pH, plasma electrolytes, and osmolality.

Results: Data obtained from seven sheep with normoxemic fetuses were studied statistically. Umbilical arterial blood flow decreased significantly from 229.5 +/- 3.83 mL/min in the control to 167.4 +/- 11.1 mL/min at 30 minutes after water infusion (P < .001). Umbilical arterial vascular resistance increased rapidly and reached its peak at approximately 60 minutes after infusion and then showed a gradual recovery to the control level (P < .001). Amniotic fluid osmolality had a high degree of correlation with umbilical arterial blood flow and vascular resistance, while fetal arterial blood pressure and heart rate had only little correlation with umbilical blood flow.

Conclusion: A distilled water infusion into the amniotic cavity in near-term pregnant sheep led to an acute drop in umbilical arterial blood flow. The changes in umbilical flow were closely correlated with those in amniotic fluid osmolality. Hemolysis in the capillary networks in the fetal membranes seems to be one of the main causes of umbilical vasoconstriction. It is speculated that the fetal membranes, including capillary networks, intramembranous pathway, and amnion epithelial cells, sense the changes in amniotic fluid osmolality, which leads to a fetal adaptation to the hypotonic environment.