p53, epidermal growth factor, and platelet-derived growth factor in uterine leiomyosarcoma and leiomyomas

Uterine leiomyosarcoma (ULMS) is an aggressive gynecological disease. Although ULMS are often found in association with benign leiomyoma (LMA), little is known regarding the relationship between these benign and malignant smooth muscle neoplasms. The objective of this study was to evaluate the expression of epidermal growth factor (EGFR), platelet-derived growth factor (PDGFR), and p53 in ULMS specimens, their prognostic relevance, and the expression of these molecular markers when compared to benign LMA specimens. Between 1991 and 2001, 25 patients were identified with high-grade primary ULMS and for whom tissue was available. Tissue microarray was created with three representative cores from each of the ULMS cases as well as from 19 patients with benign uterine leiomyomata. Immunohistochemical (IHC) staining was performed for EGFR, PDGFR, and p53. Negative and positive IHC staining was scored for each marker. Outcome analysis was performed only for ULMS. Survival was determined from the time of initial diagnosis to last follow-up. Twelve (48%) ULMS expressed p53 compared to none of the LMA (P < 0.001), and 15 (60%) ULMS cases showed PDGFR expression compared to 32% of LMA samples (P= 0.08). EGFR expression did not differ between ULMS and LMA groups. ULMS patients with p53 expression had a poorer survival compared to ULMS patients with negative expression (P= 0.07). ULMS tumor stage had the strongest association with overall survival (P= 0.05). Our study supports previous investigations indicating that p53 expression may serve as a prognostic marker for ULMS patients. The difference in PDGFR expression between ULMS and LMA demonstrated a trend toward significance. EGFR was not commonly expressed in ULMS. These uniquely expressed markers may assist in stratifying patients by survival and identify novel therapeutic markers. Clearly, further investigation is needed.     

Immunohistochemical detection of transforming growth factor-alpha, epidermal growth factor, and vascular endothelial growth factor expression in hyperstimulated rat ovary

OBJECTIVE: The aim of the present study is to figure out the immunohistochemical expression of transforming growth factor-alpha (TGF-alpha), epidermal growth factor (EGF), and vascular endothelial growth factor (VEGF) in hyperstimulated rat ovaries. METHODS: Twenty Wistar-Albino adult female rats (250-300 g) were taken into the study. The animals were randomly divided into two groups, each containing 10 rats: (i) stimulation group and (ii) control group. In the stimulation group, a stimulation regimen was administered to induce follicular maturity and ovarian hyperstimulation syndrome (OHSS) at the end using a 30-IU follicle-stimulating hormone that was administered subcutaneously for 4 consecutive days, followed by a 30-IU human chorionic gonadotropin on day 5 to induce ovulation. The rats, in the control group, received 0.2 ml of 0.9% NaCl for 5 consecutive days to mimic the conditions of the study animals. At the end of the treatment period, all rats underwent ovariectomy and the sections of ovaries were stained for the TGF-alpha, EGF, and VEGF. RESULTS: The expression of TGF-alpha, EGF, and VEGF in the endothelium, the stroma, the granulosa cells, and the corpus luteum was found to be significantly higher in the stimulated group, compared to that in the control group ( p < 0.05). CONCLUSION: TGF-alpha, EGF, and VEGF are found to have increased in the hyperstimulated ovaries and this finding seems to be involved in the OHSS pathogenesis.     

Markedly elevated levels of vascular endothelial growth factor, fibroblast growth factor, and interleukin 6 in Meigs syndrome

Analysis of serum and peritoneal and pleural fluid from a patient with Meigs' syndrome revealed high levels of vascular endothelial growth factor, fibroblast growth factor, and interleukin 6. Serum levels declined after removal of the ovarian tumor, along with resolution of ascites and hydrothorax. These findings suggest the involvement of these vasoactive factors in ascites and pleural fluid formation in Meigs' syndrome.     

Vascular endothelial growth factor, placenta growth factor and their receptors in isolated human trophoblast

The expression of the angiogenic growth factors, vascular endothelial cell growth factor (VEGF) and placenta growth factor (PIGF) was demonstrated in isolated human term cytotrophoblast and in vitro differentiated syncytiotrophoblast. RNase protection assays demonstrated VEGF expression in both cytotrophoblast and syncytiotrophoblast while prominent PIGF expression was detected in both types of trophoblast by Northern blot analyses. VEGF expression increased approximately eightfold in trophoblast cultured under hypoxic conditions (1 per cent O₂) yet PIGF expression decreased 73 ± 5.5 per cent in the same trophoblast. These results suggest distinct regulatory mechanisms govern expression of VEGF and PIGF in trophoblast. Characterization of the VEGF/PIGF receptors, KDR and flt-1, revealed the presence of flt-1 mRNA in isolated cytotrophoblast and in vitro differentiated syncytiotrophoblast. KDR was not detected in the isolated trophoblast. Exogenous rhVEGF induced c-Jun N-terminal kinase (JNK) activity in the normal trophoblast indicating that the flt-1 receptors on trophoblast are functional. Trophoblast-derived VEGF/PIGF could act in a paracrine fashion to promote uterine angiogenesis and vascular permeability within the placental bed. In addition, presence of functional flt-1 on normal trophoblast suggests that VEGF/P1GF functions in an autocrine manner to perform an as yet undefined role in trophoblast invasion, differentiation, and/or metabolic activity during placentation.     

Expression of transforming growth factor-alpha, epidermal growth factor, and epidermal growth factor receptor in follicles of human ovarian tissue before and after cryopreservation

OBJECTIVE: To study the expression of transforming growth factor-alpha (TGF-alpha), epidermal growth factor (EGF), and EGF receptor in follicles of human ovarian tissue. DESIGN: A retrospective, controlled comparative study. SETTING: In vitro fertilization laboratory of a university hospital. PATIENT(S): Fifteen women with regular menstrual cycles who underwent laparoscopy and the biopsy of ovarian tissue. INTERVENTION(S): Paraffin sections were prepared from ovarian tissues, followed by immunohistochemical staining of TGF-alpha, EGF, and EGF receptor. MAIN OUTCOME MEASURE(S): Immunostaining for TGF-alpha, EGF, and EGF receptor in follicles of fresh and frozen ovarian tissues. RESULT(S): Immunoreactivities for TGF-alpha and EGF receptor were observed simultaneously in the oocytes of primordial, primary, preantral, and antral follicles. Strong staining for TGF-alpha and EGF receptor was present in thecal cells. The TGF-alpha and EGF receptor was also expressed in some granulosa cells of primary to antral follicles. The EGF only stained weakly in the oocytes of primordial and primary follicles and in thecal cells. There was no difference in staining patterns for TGF-alpha, EGF, and EGF receptor between fresh and frozen ovarian tissues. CONCLUSION(S): The TGF-alpha and EGF receptor was expressed in primordial to antral follicles, indicating a role of TGF-alpha in regulating follicular development through binding to the EGF receptor. Freeze-thawing did not substantially alter immunoreactivites for TGF-alpha, EGF, and EGF receptor in frozen ovarian tissue.     

Keratinocyte growth factor promotes the survival, growth, and differentiation of preantral ovarian follicles

OBJECTIVE: To determine the effect of treatment with keratinocyte growth factor (KGF) on the survival of cells in cultured preantral follicles and on the growth and differentiation of preantral follicles. DESIGN: Preantral follicles (140-150 microm) were dissected mechanically from the ovaries of 14-day-old rats and cultured for 24 hours with and without KGF. Genomic DNA was extracted, labeled with [32P]-dideoxyadenosine triphosphate, and fractionated through agarose gels. For growth studies, the follicles were cultured individually in 96-well dishes. After 72 hours, the follicles were collected and their protein or DNA content was evaluated and their inhibin-alpha content was determined. RESULT(S): Keratinocyte growth factor suppressed apoptosis in cultured preantral follicles by 60%. Treatment with KGF or FSH increased follicle diameter by 8% and 16%, respectively, and combined treatment with KGF and FSH increased follicle diameter by 26%. Western blot analysis demonstrated increased expression of inhibin-alpha content after treatment with KGF (2-fold), treatment with FSH (4-fold), and combined treatment with FSH and KGF (12-fold), demonstrating the effect of KGF on preantral follicle differentiation. CONCLUSION(S): Treatment with KGF promotes the survival, growth, and differentiation of cultured preantral follicles. Keratinocyte growth factor produced by theca cells may play a role in the progression of early follicle development.     

Concentrations of alpha-fetoprotein, insulin-like growth factor binding protein-3, c-erbB-2, and epidermal growth factor in serum of patients with endometriosis

OBJECTIVE: Endometriosis, although it is a benign disorder, shares many similarities with cancer. There is increasing levels of evidence suggesting that some circulating factors involved in gynecologic cancers, such as alpha-fetoprotein (AFP), insulin-like growth factor binding protein-3 (IGFBP-3), c-erbB-2 (HER-2/neu), and epidermal growth factor (EGF), could also play a role in endometriosis. Hence, the present study was aimed at evaluating whether the levels of these molecules are modulated in the serum of patients with endometriosis. METHODS: Levels of AFP, IGFBP-3, c-erbB-2, and EGF were determined by enzyme-linked immunosorbent assay in serum from 36 subjects with surgically confirmed endometriosis and 36 controls with no surgical evidence of the disease. In addition, information such as demographic characteristics, personal habits, menstrual characteristics, and clinical profile was collected from each participating subject. RESULTS: No significant difference was found between serum levels of AFP, IGFBP-3, c-erbB-2, and EGF in patients with endometriosis and controls, even when we adjusted for potential confounders and took into account the menstrual cycle. Moreover, no correlation was observed between the serum concentrations of these molecules and the stage of the disease. However, a correlation was detected between soluble levels of IGFBP-3 and presence of uterine leiomyoma. CONCLUSION: Although AFP, IGFBP-3, c-erbB-2, and EGF are not altered in the circulation of patients with endometriosis, their involvement in the development of endometriotic lesions cannot be excluded.     

Second-trimester maternal serum placental growth factor and vascular endothelial growth factor for predicting severe,early-onset preeclampsia

OBJECTIVE: To determine whether alterations in second-trimester maternal serum cytokine concentrations can identify women at risk for developing severe, early-onset preeclampsia. METHODS: Patients with severe preeclampsia requiring delivery prior to 34 weeks (n = 20) were each matched by gestational age, gravidity, parity, and sample freezing time with three healthy controls who delivered at term (n = 60). By using second-trimester maternal sera originally collected for fetal aneuploidy screening, the concentrations of placental growth factor, vascular endothelial growth factor, granulocyte colony-stimulating factor, endothelin-1, and human chorionic gonadotropin were compared between patients and controls. Logistic regression analysis was used to estimate odds ratios for high versus low (median split) cytokine concentrations with respect to the development of severe, early-onset preeclampsia. Receiver operating characteristic (ROC) curves based on a second logistic regression, using actual cytokine values, were plotted to illustrate reciprocal impact on sensitivity and specificity. RESULTS: Placental growth factor and vascular endothelial growth factor levels were significantly lower in patients than in controls. No significant differences were observed for the other cytokines. The odds ratios (with 95% confidence intervals) were 15.54 (3.29, 73.40) for vascular endothelial growth factor and 4.20 (1.35, 13.06) for placental growth factor. Receiver operating characteristic analysis of placental growth factor and vascular endothelial growth factor confirmed that both were useful in discriminating between patients and controls. Models combining both vascular endothelial growth factor and placental growth factor provided the best performance for identifying patients at risk for developing severe, early-onset preeclampsia, according to both odds ratios and ROC analyses. CONCLUSION: Combined analysis of placental growth factor and vascular endothelial growth factor is potentially useful as a tool for early identification of patients at risk for developing severe, early-onset preeclampsia.     

Expression of epidermal growth factor receptors and epidermal growth factor,amphiregulin and neuregulin in bovine uteroplacental tissues during gestation

Introduction

Growth factors are proteins that bind to specific cell surface receptors that initiate signalling pathways and result in proliferation or differentiation of the affected cells. During gestation, epidermal growth factor receptors (ErbB1-4) and its ligands (epidermal growth factor-EGF, amphiregulin-AREG, neuregulin1-NRG1) play a significant role in differentiation, function and growth of the uterus.

Objectives

To determine the role of ErbB receptors and EGF, AREG and NRG1 in bovine uteroplacental tissues during gestation.

Methods

Placentomes and interplacentomal areas from 30 cows from early gestation until near term were analysed by immunohistochemistry.

Results

ErbB receptors and its ligands were observed in uteroplacental tissues and its expression was maintained throughout pregnancy, but ErbB1 receptor did not exist in the caruncular and cotyledonary stromal cells. Besides, caruncular stromal cells did not present with any immune reaction for EGF, AREG and NRG1. Generally, it was observed that total scores for ErbB receptors and its ligands (EGF, AREG and NRG1) had decreased from early to late gestation (p < 0.05).

Conclusion

Our study shows the presence of ErbB receptors and its ligands participate in the mid- and late-phases of pregnancy. To our knowledge, this is the first study on the expression of NRG1 during bovine pregnancy. These results indicates that these factors may play a crucial role not only to enable cellular proliferation and differentiation in the uterus throughout gestation, but also to have a potential role in the cellular communication maintained between the embryo/fetus and uterus by the placenta.     

胰岛素样生长因子1及胰岛素样生长因子结合蛋白3与胎儿生长发育的关系

目的:探讨胰岛素样生长因子1（IGF-1）及胰岛素样生长因子结合蛋白3（IGFBP-3）与胎儿生长发育的关系。方法:应用酶联免疫吸附试验（LISA）测定26例正常妊娠（正常组）,42例妊娠期糖尿病（GDM组）,20例胎儿宫内发育迟缓（IUGR组）孕妇足月剖宫产分娩时,母血与脐血中IGF-1及IGFBP-3的水平,同时记录3组孕妇的新生儿出生体重。结果:（1）母血IGF-1及IGFBP-3的水平正常组分别为18 6.81μg/L、22.82μg/L,GDM组为283.35μg/L、28.29μg/L,IUGR组为220.64μg/L、25.23μg/L,3组间 IGF-IN IGFBP.3水平差异均无显著性（P>0.05）;（2）脐血IGF-1及IGFBP-3的水平正常组分别为62.54μg/L、8.56μg/L,GDM组分别为83.74μg/L、10.21μg/L,IUGR组为37.94μg/L、7.82μg/L,分别进行3组间两两比较,3组IGF-1及IGFBP-3的差异均有显著性（P<0.01）;（3）新生儿平均出生体重正常组为3.22±0.32kg,GDM组为3.76±0.43kg,IUGR组为2.41±0.17kg,3组间两两比较,差异均有显著性（P<0.01）;（4）3组脐血IGF-1及IGFBP-3水平与新生儿出生体重均有显著性正相关（P<0.01）;（5）3组母血及脐血的IGF-1与IGFBP-3均呈显著性正相关（P<0.01）。结论:来自胎儿循环的IGF-1、IGFBP-3对胎儿的生长发育有重要的调节作用,可能参与巨大儿及IUGR的病     

Expression of von Willebrand's factor,CD34, CD31, and vascular endothelial growth factor in uterine leiomyomas

OBJECTIVE: To compare the vascular parameters of uterine leiomyomas and normal myometrium, to correlate these parameters with vascular endothelial growth factor (VEGF) expression and clinical/pathological parameters, and to compare vascular parameters according to the endothelial markers used.DESIGN: An immunohistochemical technique was applied to formalin-fixed paraffin-embedded tissue samples, using antibodies against von Willebrand's factor (FvW), CD34, CD31, and VEGF. The intratumoral vascular area (VA), microvessel density (MVD), and vascular luminal area (VLA) were determined with an image analyser.SETTING: University teaching hospital.PATIENT(S): Thirty-two patients with uterine leiomyomas underwent conservative surgery. Twenty leiomyoma-free patients undergoing hysterectomy were the controls.INTERVENTION(S): Immunohistochemical and morphometrical analysis.MAIN OUTCOME MEASURE(S): Measurements of VA, MVD, and VLA.RESULT(S): The CD34 labeling showed decreased VA in myomas compared with myometrium. Decreased MVD and an increased VLA in myomas were found with FvW and CD34 labeling. The VA, MVD, and VLA were not related to VEGF expression or to clinical/pathological parameters. Similar results for VA and MVD were obtained with FvW and CD34 labeling.CONCLUSION(S): Leiomyomas have a smaller vascular area, a lower microvessel density, and a higher vascular luminal area than normal myometrium.     

Reductions of vascular endothelial growth factor and placental growth factor concentrations in severe preeclampsia

OBJECTIVE: The aim of this study was to determine whether plasma concentrations of vascular endothelial growth factor and placental growth factor are altered in women with severe preeclampsia. STUDY DESIGN: We performed a case-control study to compare plasma concentrations of vascular endothelial growth factor and placental growth factor between women with severe preeclampsia and normotensive women admitted for delivery. Twenty-one women with severe preeclampsia were matched for gestational age and ethnicity with 21 normotensive women. Vascular endothelial growth factor and placental growth factor concentrations were measured with a specific antigen-capture enzyme-linked immunosorbent assay. RESULTS: Women with severe preeclampsia demonstrated significantly lower plasma concentrations of both vascular endothelial growth factor (6.36 +/- 3.96 pg/mL vs 18.65 +/- 5.98 pg/mL; P <.0001) and placental growth factor (138 +/- 119 pg/mL vs 531 +/- 340 pg/mL; P <.0001) than did women with normotensive pregnancy. Logistic regression analysis showed an independent association between plasma vascular endothelial growth factor concentration and plasma placental growth factor concentration and preeclampsia. CONCLUSION: Patients with severe preeclampsia had decreased maternal serum concentrations of both vascular endothelial growth factor and placental growth factor.     

Preeclampsia: the endothelium, circulating factor(s) and vascular endothelial growth factor

It has been proposed that endothelial cell activation is the primary event in the multisystem disorder of preeclampsia. Evidence for endothelial involvement in this condition abounds. The best-characterized morphologic abnormality of this syndrome, glomerular endotheliosis, involves endothelial cells. Also associated with preeclampsia is a loss of endothelial cell integrity, with the consequent increase in vascular permeability, and an increase in the circulating levels of the endothelial cell markers, fibronectin, von Willebrand factor, tissue plasminogen activator, and plasminogen activator inhibitor-1. It is now well documented that endothelial activation contributes to the coagulation abnormalities observed in this disease. There is much evidence that the endothelial alterations in preeclampsia result from one or more circulating factors. The incubation of cultured endothelial cells with serum or plasma samples, taken from normal pregnant women and women with preeclampsia, results in marked alterations in cell behavior and metabolic processes. More recently, experiments employing myographic techniques have demonstrated convincingly the effects of a circulating factor(s) on the function of endothelial cells of resistance arteries. Vascular endothelial growth factor (VEGF) possesses many of the characteristics required of a candidate circulating factor. It contains a hydrophobic secretory signal sequence, exerts in vitro effects specific to vascular endothelial cell, and promotes endothelial expression of procoagulant activity. Circulating VEGF concentrations are elevated in women with preeclampsia, and VEGF increases microvascular endothelial cell prostacyclin production in a dose-dependent manner, analogous to the acute effects of plasma from patients with preeclampsia. Similarly, in myographic studies, when myometrial resistance arteries are incubated with VEGF, there are dose-dependent alterations in endothelium-dependent behavior, mirroring those found after incubation with plasma from patients with preeclampsia.     

Binding of epidermal growth factor and insulin-like growth factor I in human myometrium and leiomyomata

Samples of uterine myometrium and leiomyoma from 11 women were analyzed for the presence of epidermal growth factor receptors and insulin-like growth factor I receptors. In addition, the content of soluble insulin-like growth factor binding protein (IGF-BP/PP12) was measured in the tissue cytosols. Cell membrane preparations of myoma tissue bound significantly more insulin-like growth factor I than did those of adjacent normal myometrium, whereas myoma tissue bound less epidermal growth factor than did the normal myometrium. The differences in both insulin-like growth factor I and epidermal growth factor binding were due to changes in receptor concentration rather than to alterations in receptor affinity. Neither myoma nor myometrial tissue contained detectable levels of insulin-like growth factor binding protein. The changes in epidermal growth factor and insulin-like growth factor I binding to the myometrium may play a role in the pathogenesis of uterine leiomyomata.