epidermal-growth-factor and Leiomyoma

Topics: Diphosphonates; Drug Therapy, Combination; Epidermal Growth Factor; ErbB Receptors; Estrogens; Female; Gonadotropin-Releasing Hormone; Humans; Leiomyoma; Leuprolide; Uterine Neoplasms

It is now evident that the use of levonorgestrel-releasing intrauterine system (LNg-IUS) is effective for long-term management of menorrhagic women with uterine myomas because of a striking reduction in menorrhagia. This prompted us to characterize the effects of progesterone (P4) on the growth and apoptosis of uterine leiomyoma cells. On the other hand, we have recently noted that epidermal growth factor (EGF) and IGF-I play a crucial role in prompting uterine leiomyoma growth through stimulating the proliferative potential and inhibiting apoptosis of cultured human leiomyoma cells. In the present review, attention was paid to evaluate the effects of P4 on the expression of growth factors (EGF, IGF-I) and apoptosis-related factors (TNFalpha, Bcl-2 protein) in cultured uterine leiomyoma cells. Treatment with P4 augmented EGF and Bcl-2 protein expression, but inhibited IGF-I and TNFalpha expression in cultured leiomyoma cells. It is known that TNFalpha induces apoptosis in a variety of cell types and Bcl-2 protein is an apoptosis-inhibiting gene product. Thus, the results obtained suggest that P4 has dual actions on uterine leiomyoma growth: one is to stimulate leiomyoma cell growth and survival through up-regulating EGF and Bcl-2 protein expression as well as down-regulating TNFalpha expression in those cells, and the other is to inhibit leiomyoma cell growth through down-regulating IGF-I expression in those cells. This may explain why the size of uterine myomas during use of LNg-IUS increases in some but decreases in other instances. This may also explain why the size of uterine myomas during pregnancy does not increase despite the overwhelming increase in circulating concentrations of sex steroid hormones.

Topics: Apoptosis; Blotting, Western; Cell Division; Cell Line, Tumor; Dose-Response Relationship, Drug; Down-Regulation; Epidermal Growth Factor; Female; Growth Substances; Humans; Insulin-Like Growth Factor I; Leiomyoma; Progesterone; Proto-Oncogene Proteins c-bcl-2; Tumor Necrosis Factor-alpha; Up-Regulation; Uterine Neoplasms

Recent investigations, using DNA technology, of the molecular biology of smooth muscle tumours of the uterus have confirmed their monoclonality and have strengthened the view that oestrogen and oestrogen receptors play a major role in the pathogenesis of fibromyomata. In addition, increasing evidence suggests that progesterone, insulin-like growth factors, epidermal growth factors and other proteins are also involved. The mechanisms whereby gonadotrophin-releasing hormone agonists cause shrinkage of fibromyomata remain controversial but both vascular changes and cellular atrophy appear to play a role. A shift of emphasis in the study of fibromyomata has resulted from the demonstration that the myometrium adjacent to fibromyomata is not normal and shows some similarities to the tumours themselves.

Topics: Cytogenetics; Epidermal Growth Factor; Estrogens; Female; Humans; Leiomyoma; Molecular Biology; Progesterone; Receptors, Estrogen; Somatomedins; Uterine Neoplasms

To study the effects of mifepristone on epidermal growth factor gene expression in human uterine leiomyoma.. 20 patients with leiomyoma were divided into two groups. One was control group who underwent hysterectomy because of leiomyoma, the other was experimental group who underwent hysterectomy after pretreatment with mifepristone 10 mg/daily for 3 months. Epidermal growth factor (EGF) mRNA was semiquantified in samples of leiomyoma and adjacent normal myometrium from patients untreated in different phases of menostrual cycle and from those treated with mifepristone. Semiquantitative reverse transcriptase polymerase chain reaction, using beta-actin as internal standard, was applied to determine levels of EGF mRNA.. Leiomyoma untreated with mifepristone had significantly greater amounts of EGF mRNA than adjacent normal myometrium of uterus only in the luteal phase, but not in the follicular phase of cycle. Similarly, leiomyoma untreated with mifepristone also had significantly larger amount of EGF mRNA than treated leiomyoma in the luteal phase of the cycle, whereas, no difference in the follicular phase of the cycle.. These findings suggested that: (1) EGF mRNA levels in leiomyoma were increased only in the luteal phase, therefore, maybe mainly controlled by progesterone; (2) mifepristone inhibited EGF gene expression in leiomyoma. This may be one of regression mechanism of uterine leiomyoma in response to the antiprogesterone mifepristone.

Topics: Epidermal Growth Factor; Female; Gene Expression; Hormone Antagonists; Humans; Leiomyoma; Mifepristone; RNA, Messenger; Uterine Neoplasms

Uterine leiomyomas are the most common benign tumours in women, which arise from smooth muscle cells of the uterine myometrium and usually are multicentric. In spite of their frequency pathogenesis is widely unknown, mainly due to the absence of a suitable model system. We describe the systematic optimization of culturing leiomyoma tissue explants in an economical and effective ex vivo system.. Different concentrations of oxygen, different media, sera, hormones, and growth factor supplements were tested. Immunohistochemical stainings with antibodies against hormone receptors as well as specifying proliferation and apoptotic indices and real-time PCR were performed.. Main parameters for culturing myoma tissue explants were tested for finding an optimal protocol. Standard medium D-MEM-F12 in combination with the use of horse serum in a reduced concentration of 1% turned out to be optimal for these tissue cultures as well as the addition of estradiol and epidermal growth factor EGF to media. Reduced oxygen content in the incubator air showed no positive effect.. For culturing tissue explants of uterine leiomyoma several conditions were optimized. The established tissue culture model allows examining the effects of known and potential therapeutic substances and the influence of immune competent cells in the process of tumour formation to find new targets for medical treatment.

Topics: Culture Media; Epidermal Growth Factor; Estradiol; Female; Humans; Immunohistochemistry; Leiomyoma; Progesterone; Real-Time Polymerase Chain Reaction; RNA, Messenger; Uterine Neoplasms

Investigation of activin-A (A) and myostatin (M) in human myometrium (HM) and leiomyoma (HL) will explain their involvement in human myometrial pathophysiology.. We aimed to investigate A and M response and steroid regulation in HM. We also evaluated A and M expression and response in HL.. Tissues were analyzed and cultured.. Patients included fertile (in proliferative phase) and menopausal women undergoing hysterectomy.. HM explant cultures were treated with A and M (for Smad-7 mRNA quantification) or estrogen and progesterone (for A and M mRNA quantification). A and M expression levels were also evaluated in menopausal (physiological absence of steroids) HM specimens. A and M and their receptors were evaluated in HL (n = 8, diameter 5-8 cm) compared with their matched HM. HL explants cultures were treated with A and M (for Smad7 mRNA quantification), and, to explain the absence of response, the levels of follistatin, follistatin-related gene (FLRG), and Cripto were evaluated.. A and M increased Smad7 expression in HM explants. A and M mRNAs were both reduced after estradiol treatment, unchanged after progesterone treatment, but were higher in menopausal than fertile (in proliferative phase) specimens. A, M, and FLRG were expressed at higher levels in HL compared with adjacent HM, whereas the receptors, follistatin, and Smad7 mRNAs resulted unchanged. Cripto mRNA was expressed only in HL.. A and M act on human HM and are regulated by steroids. In HL there is an increase of A, M, FLRG, and Cripto expression.

Topics: Activin Receptors; Activins; Adult; Blotting, Western; Endometrium; Epidermal Growth Factor; Female; Follistatin; Follistatin-Related Proteins; Gene Expression Regulation; GPI-Linked Proteins; Humans; Intercellular Signaling Peptides and Proteins; Leiomyoma; Luciferases; Membrane Glycoproteins; Middle Aged; Myometrium; Myostatin; Neoplasm Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA; Signal Transduction; Smad7 Protein; Steroids; Tissue Culture Techniques

To determine different effects of epidermal growth factor (EGF) on cultured smooth muscle cells (SMCs) derived from human myometrium and leiomyoma.. EGF effects on DNA synthesis and intracellular signal transduction were studied in cultured SMCs from leiomyoma and its matched myometrium.. Research laboratories.. Patients 35-50 years old with uterine leiomyomas.. Hysterectomy.. Signal transduction from EGF receptor.. As analyzed by laser scanning cytometry (LSC), EGF treatment stimulated DNA synthesis and induced polyploidization of leiomyomal, but not myometrial, SMCs. EGF stimulation was inhibited by AG1478, an EGF receptor (EGFR) inhibitor and PD98059, a mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor. Both leiomyomal and myometrial SMCs had similar expression levels of EGFR, but EGF treatment induced transient phosphorylation activation of EGFR and Akt in leiomyomal SMCs. Consequently, EGF triggered transient phosphorylation activation of p44/42 MAPK in leiomyomal SMCs, followed by down-regulation of p27. In myometrial SMCs, however, EGF induced sustained activation of EGFR, Akt, and p44/42 MAPK with up-regulation of p27.. EGF stimulates DNA synthesis and polyploidization in leiomyomal SMCs through transient activation of the EGFR-MAPK pathway. Given that polyploidization plays a role in tumorigenesis, our results shed new light on the pathogenesis of human uterine leiomyoma.

Topics: Adult; Cells, Cultured; Epidermal Growth Factor; Female; Humans; Leiomyoma; Middle Aged; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Myometrium; Polyploidy

Uterine leiomyomas are benign uterine tumors characterized by extracellular matrix remodeling, increased collagen deposition, and increased smooth muscle cell (SMC) proliferation. The reactive oxygen species (ROS) producing NADPH oxidase complex has been shown to be involved in the signaling pathways of several growth factors, cytokines, and vasoactive agents that stimulate proliferation of a variety of cell types. Our objective was to test the hypothesis that ROS derived from NADPH oxidase is a necessary component of the MAP kinase mitogenic pathway activated by platelet derived growth factor (PDGF) and epidermal growth factor (EGF) in leiomyoma SMCs (LSMCs). Primary cell cultures of LSMCs were used as our experimental model. Our results showed that stimulation of these cells with PDGF or EGF caused a marked increase in intracellular ROS production and that the NADPH oxidase inhibitor, DPI, blocks ROS production. In addition, inhibition of ROS production by NADPH oxidase inhibitors blocked, in a dose-dependent manner, the EGF- and PDGF-induced increase in [(3)H]thymidine incorporation by LSMCs. Furthermore, an exogenous source of ROS, hydrogen peroxide, was sufficient to stimulate [(3)H]thymidine incorporation in LSMCs but did not affect COL1A2 and COL3A1 mRNA levels. Inhibition of the NADPH oxidase complex decreased PDGF-induced MAPK1/MAPK3 activation, whereas exogenous hydrogen peroxide induced MAPK1/MAPK3 activation. This article is the first report suggesting the presence of the NADPH oxidase system and its importance in mitogenic signaling pathways in LSMCs. The necessity of NADPH oxidase-derived ROS for EGF and PDGF signaling pathways leading to cell proliferation points to another potential therapeutic target for treatment and/or prevention of uterine leiomyomas.

Topics: Cell Division; Cell Line, Tumor; DNA; Enzyme Activation; Enzyme Inhibitors; Epidermal Growth Factor; Female; Humans; Hydrogen Peroxide; Leiomyoma; Mitogen-Activated Protein Kinases; Muscle, Smooth; NADPH Oxidases; Platelet-Derived Growth Factor; Reactive Oxygen Species; Signal Transduction; Uterine Neoplasms

Human uterine leiomyomas are very common smooth muscle cell tumors that occur in reproductive-age women and are the leading reason for performing hysterectomies. The present study was conducted to explore the potential mechanism behind the effects exerted by dominant-negative estrogen receptors (DNERs) delivered by adenovirus to leiomyoma cells to ascertain the utility of DNERs as a novel strategy for treatment of uterine fibroids.. We investigated the ability of DNER to affect estrogen response element (ERE) activity induced by wild-type estrogen receptor (ER) by using the adenovirus ERE luciferase (Ad-ERE-luc) system in ELT3 cells and the effect of graded doses of DNER (10, 50, and 100 plaque-forming units/cell) on the expression of some selected genes controlling cultured human leiomyoma cell proliferation (cyclin D1, Cox2, PCNA, VEGF, and EGF), apoptosis (Bcl2 and Bax), estrogen metabolism (COMT), and extracellular matrix formation (MMP(1)) as well as progesterone receptors (A and B) were assessed using Western blot analysis. These genes are all regulated by estrogen and/or progesterone.. DNER has the ability to suppress the ERE luc activity induced by wild-type ER (P < 0.01) and significantly (P < 0.05) reverse the expression of all estrogen- and progesterone-regulated genes in this study.. These results suggest that interruption of the estrogen signaling pathway using DNER results in modulation of both estrogen- and progesterone-regulated genes that control leiomyoma cell apoptosis, proliferation, extracellular matrix formation, progesterone receptors, and estrogen metabolism, which might account for the DNER mechanism of action.

Topics: Adenoviridae; Animals; Apoptosis; Catechol O-Methyltransferase; Cell Line, Tumor; Down-Regulation; Epidermal Growth Factor; Estrogens; Female; Gene Expression Regulation, Neoplastic; Genes, Dominant; Genetic Therapy; Leiomyoma; Matrix Metalloproteinase 1; Progesterone; Rats; Receptors, Estrogen; Receptors, Progesterone; Signal Transduction; Transcriptional Activation; Transfection; Up-Regulation; Uterine Neoplasms; Vascular Endothelial Growth Factor A

Progesterone plays a pivotal role in controlling uterine leiomyoma growth. The authors review studies they conducted to evaluate the comparative effects of asoprisnil on proliferation, apoptosis, and growth factor expression in cultured leiomyoma and normal myometrial cells. Treatment with asoprisnil decreased the proliferating cell nuclear antigen-positive rate and the number of viable cells and increased the terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end labeling- positive rate in cultured leiomyoma cells in a dose-dependent manner ( P < .05). Similarly, asoprisnil decreased Bcl-2 expression and increased cleaved caspase-3 and cleaved poly(adenosine 5'-diphosphate-ribose) polymerase in leiomyoma cells but not in normal myometrial cells. Similarly, asoprisnil decreased epidermal growth factor (EGF), insulin-like growth factor-I (IGF-I), and transforming growth factor (TGF) beta mRNA and protein expression, as well as EGF receptor, IGF-IR alpha, and TGF RII protein expression in leiomyoma cells but not in cultured normal myometrial cells. These results suggest that asoprisnil selectively inhibits proliferation by downregulating the growth factors and their receptor expression and induces apoptosis in leiomyoma cells without affecting proliferation and apoptosis in normal myometrial cells.

Topics: Apoptosis; Cell Proliferation; Cell Survival; Cells, Cultured; Dose-Response Relationship, Drug; Down-Regulation; Epidermal Growth Factor; ErbB Receptors; Estrenes; Female; Humans; Insulin-Like Growth Factor I; Intercellular Signaling Peptides and Proteins; Leiomyoma; Myometrium; Oximes; Phosphorylation; Progesterone; Proliferating Cell Nuclear Antigen; Protein Serine-Threonine Kinases; Receptor, IGF Type 1; Receptor, Transforming Growth Factor-beta Type II; Receptors, Progesterone; Receptors, Transforming Growth Factor beta; RNA, Messenger; Time Factors; Transforming Growth Factor beta3; Tumor Cells, Cultured; Uterine Neoplasms

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.. 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.. 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.. Asoprisnil inhibits the expression of EGF, IGF-I, TGFbeta3 and their receptors in cultured leiomyoma cells without affecting their expressions in myometrial cells.

Topics: Adult; Blotting, Western; Cell Proliferation; Cells, Cultured; Down-Regulation; Epidermal Growth Factor; ErbB Receptors; Estrenes; Female; Gene Expression; Humans; Insulin-Like Growth Factor I; Leiomyoma; Middle Aged; Myometrium; Oximes; Proteoglycans; Receptor, IGF Type 1; Receptors, Growth Factor; Receptors, Progesterone; Receptors, Transforming Growth Factor beta; Reverse Transcriptase Polymerase Chain Reaction; Transforming Growth Factor beta; Tumor Cells, Cultured; Uterine Neoplasms

To investigate the role that the hedgehog (Hh) signaling pathway, which includes sonic hedgehog (Shh), Patched (Ptc), Smoothened (Smo) and Gli-1, plays in human gastrointestinal stromal tumors (GISTs).. Surgically resected specimens from patients with GISTs, leiomyomas and schwannomas were examined by immunohistochemical staining for aberrant expression of hedgehog signaling components, Shh, Ptc, Smo and Gli-1, respectively.. In GISTs, 58.1% (18 of 31), 77.4% (24 of 31), 80.6% (25 of 31) and 58.1% (18 of 31) of the specimens stained positive for Shh, Ptc, Smo and Gli-1, respectively. In leiomyomas, 92.3% (12 of 13), 92.3% (12 of 13), 69.2% (9 of 13) and 92.3% (12 of 13) stained positive for Shh, Ptc, Smo and Gli-1, respectively. In schwannomas, 83.3% (5 of 6), 83.3% (5 of 6), 83.3% (5 of 6) and 100% (6 of 6) stained positive for Shh, Ptc, Smo and Gli-1, respectively. Immunohistochemistry revealed that the expressions of Shh and Gli-1 were significantly higher in leiomyomas than in GISTs (P < 0.05, respectively). Shh expression strongly correlated with the grade of tumor risk category and with tumor size (P < 0.05, respectively). However, the expressions of Ptc and Smo did not correlate with histopathological differentiation.. These results suggest that the Hh signaling pathway may play an important role in myogenic differentiation and the malignant potential of human intestinal stromal tumors.

Topics: Cell Transformation, Neoplastic; Epidermal Growth Factor; Gastrointestinal Stromal Tumors; Gene Expression Regulation, Neoplastic; Hedgehog Proteins; Humans; Intestinal Neoplasms; Leiomyoma; Neurilemmoma; Patched Receptors; Prognosis; Protein-Tyrosine Kinases; Receptors, Cell Surface; Receptors, G-Protein-Coupled; Signal Transduction; Smoothened Receptor; Transcription Factors; Zinc Finger Protein GLI1

The objective of this study was to elucidate the effects of GnRH antagonist Cetrorelix on proliferation and apoptosis in human leiomyoma cells cultured in vitro. Isolated leiomyoma cells were subcultured in phenol red-free DMEM supplemented with 10% fetal bovine serum for 120 h and then stepped down to serum-free conditions in the presence or absence of graded concentrations of Cetrorelix (10(-5) to 10(-8) mol/liter) for 6 d. Cultured leiomyoma cells were used for semiquantitative RT-PCR, immunocytochemistry, Western blot analysis, and terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling assay. RT-PCR analysis revealed the presence of mRNAs encoding for GnRH receptor and epidermal growth factor (EGF) in cultured leiomyoma cells. The number of viable cultured leiomyoma cells was significantly (P < 0.01) decreased by treatment with Cetrorelix compared with untreated control cultures. Immunocytochemical examination demonstrated that treatment with Cetrorelix attenuated the expression of proliferating cell nuclear antigen (PCNA) and EGF in cultured leiomyoma cells. Western blot analysis revealed that treatment with 10(-5) mol/liter Cetrorelix significantly (P < 0.01) decreased PCNA expression. In addition, treatment with 10(-5) mol/liter Cetrorelix remarkably increased the terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling-positive rate and poly(ADP-ribose) polymerase expression at 24 h of treatment compared with untreated control cultures (P < 0.01). Furthermore, treatment with 10(-5) mol/liter Cetrorelix decreased immunoreactive EGF protein and EGF mRNA expression in cultured leiomyoma cells at 4 d of treatment. GnRH antagonist Cetrorelix may directly inhibit leiomyoma cell growth by down-regulating proliferation in association with a decrease in EGF mRNA expression and by up-regulating apoptosis in those cells.

Topics: Adult; Apoptosis; Base Sequence; Cell Survival; DNA Primers; Epidermal Growth Factor; Female; Gene Expression Regulation, Neoplastic; Gonadotropin-Releasing Hormone; Humans; Leiomyoma; Poly(ADP-ribose) Polymerases; Premenopause; Proliferating Cell Nuclear Antigen; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured; Uterine Neoplasms

The objective of this study was to investigate the comparative effects of heparin-binding epidermal growth factor-like growth factor (HB-EGF) on the growth of cultured human leiomyoma cells and myometrial cells.. Isolated cells were subcultured in Phenol Red-free Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum for 120 h and then stepped down to serum-free conditions for an additional 24 and 48 h in the presence or absence of graded concentrations of HB-EGF (0.1, 1, 10 and 100 ng/ml). These cells were used for immunocytochemical analysis for Ki67, western blot analysis for proliferating cell nuclear antigen (PCNA) and human EGF receptor (HER1), and TdT (terminal deoxynucleotidyl transferase)-mediated dUDP nick-end labelling (TUNEL) assay.. Treatment with HB-EGF at concentrations >1 ng/ml significantly increased the Ki67-positive rate of cultured leiomyoma cells and myometrial cells. Treatment with HB-EGF also resulted in a dose-dependent increase in PCNA expression in both cells compared with untreated control cultures. A significant increase in PCNA expression in cultured myometrial cells was noted following treatment with HB-EGF at concentrations >1 ng/ml, whereas an increase in PCNA expression in cultured leiomyoma cells was noted following treatment with HB-EGF at concentrations >10 ng/ml. HER1 expression was significantly higher in untreated myometrial cells than in untreated leiomyoma cells. A significant increase in HER1 expression in myometrial cells was observed when treated with HB-EGF at concentrations >10 ng/ml, whereas a significant increase in HER1 expression in leiomyoma cells was noted only by the treatment with HB-EGF at concentrations >100 ng/ml. Treatment with HB-EGF decreased the TUNEL-positive rate of those cells with no significant differences between the two cell types.. The results obtained suggest that HB-EGF plays a role in stimulating the proliferation of leiomyoma cells and myometrial cells and in inhibiting apoptosis of those cells through augmentation of HER1 expression. Since the proliferative potential of myometrial cells responded better to HB-EGF than that of leiomyoma cells, HB-EGF may play a more vital role in myometrial growth than leiomyoma growth.

Topics: Adult; Apoptosis; Cell Proliferation; Cells, Cultured; Deoxyuracil Nucleotides; Epidermal Growth Factor; ErbB Receptors; Female; Heparin-binding EGF-like Growth Factor; Humans; In Situ Nick-End Labeling; Intercellular Signaling Peptides and Proteins; Ki-67 Antigen; Leiomyoma; Myometrium; Proliferating Cell Nuclear Antigen; Uterine Neoplasms

Extracellular matrix is a place where various growth factors are bound and immobilised. It is expected that leiomyoma-associated remodelling of extracellular matrix in the uterus may be evoked by changes in the content of some growth factors.. The amount and distribution of EGF in the normal myometrium and in the leiomyoma during various growth stages were investigated.. The assay of EGF was carried out with the use of ELISA commercial kit. SDS/polyacrylamide gel electrophoresis of tissue extracts followed by Western immunoblot was performed.. It was found that all investigated tissues contained endogenous EGE Extractability of EGF depended on type of extracting solvent. Only slight amount of EGF could be extracted by 1 M acetic acid. Much more EGF could be solubilized in 0.05M Tris/HCl, pH 7.6. Our results showed that EGF bound to the uterus (normal and leiomyoma) components of different molecular mass. It is of interest that some components of both, acidic and neutral extracts (myometrium and leiomyomas) were not able to bind exogenous 125I-labelled EGF.

Topics: Adult; Blotting, Western; Epidermal Growth Factor; Extracellular Matrix; Female; Humans; Immunohistochemistry; Leiomyoma; Middle Aged; Myometrium; Uterine Neoplasms

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.. 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.. 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.. 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.

Topics: Adolescent; Adult; alpha-Fetoproteins; Endometriosis; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Female; Humans; Insulin-Like Growth Factor Binding Protein 3; Leiomyoma; Receptor, ErbB-2; Uterine Neoplasms

Immunolocalization of transforming growth factor alpha (TGF-Alpha), epidermal growth factor (EGF), insulinlike growth factor (IGF)-I, vascular endothelial growth factor (VEGF(165,189,121)), basic fibroblast growth factor (FGF)-2, EGF receptor (R), IGF-IRbeta, and FGFR-1 was studied in uterine leiomyomas and matched myometrial samples taken from seven women (42-47 years of age) in the proliferative phase of the menstrual cycle. Immunolocalization of growth factor peptides was accomplished with either monoclonal or polyclonal antibodies to the amino or carboxy terminus of growth factor peptides or their respective receptors, or against full-length recombinant growth factor. All reactions were conducted using the avidin-biotin complex method. Immunolocalization of TGF-alpha, EGF, EGF-R, IGF-I, IGF-IRbeta, FGF-2, FGFR-1, and VEGF was observed in the cytoplasm of smooth-muscle cells of leiomyomas and matched myometrium. The cytoplasm of vascular smooth-muscle cells expressed TGF-alpha, EGF, EGF-R, IGF-I, IGF-IRbeta, FGF-2, FGFR-1, and VEGF, whereas the vascular endothelium was positive for TGF-alpha, EGF, EGF-R, FGF-2, and FGFR-1 in both leiomyomas and matched myometria. Fibroblasts within the fibrous component of some leiomyomas were positive for IGF-I and FGF-2 and minimally positive for FGFR-1. In addition, the extracellular matrix of leiomyomas showed focal localization of FGF-2 and IGF-I in some tumors. When scores of intensity and percent positive staining were compared, IGF-IRbeta was significantly increased in the leiomyomas compared to matched myometria, whereas EGF was significantly decreased in the uterine leiomyomas compared to matched myometria. In summary, these data revealed growth factors to be expressed differentially in smooth muscle, vascular and fibroblastic cell types of leiomyomas and matched myometria. Specifically, IGF-IRbeta was significantly increased in leiomyomas; although a similar increase was seen with IGF-I peptide, statistical significance was not achieved. The EGF peptide was significantly decreased in the leiomyomas compared to matched myometrium. These data suggest that IGF-IRbeta and IGF-I peptide may be one of several growth factor/receptor pathways important in uterine leiomyoma growth during the proliferative phase of the menstrual cycle. In addition, decreased EGF may be secondary to the predominant estrogenic milieu present at time of sampling, as it has been proposed that progesterone, and not estrogen, may regulate E

Topics: Adult; Case-Control Studies; Endothelial Growth Factors; Epidermal Growth Factor; ErbB Receptors; Female; Fibroblast Growth Factor 2; Growth Substances; Humans; Immunohistochemistry; Leiomyoma; Lymphokines; Menstrual Cycle; Middle Aged; Receptor Protein-Tyrosine Kinases; Receptor, Fibroblast Growth Factor, Type 1; Receptor, IGF Type 2; Receptors, Fibroblast Growth Factor; Receptors, Growth Factor; Somatomedins; Transforming Growth Factor alpha; Uterine Neoplasms; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

To study the effects of epidermal growth factor (EGF) on uterine leiomyoma and the relationship of EGF and estrogen, progesterone.. Fourty patients with uterine leiomyoma and twenty normal women were studied. Serum EGF level was determined by radioimumunoassay, estradiol (E2) and progesterone (P) levels were measured by enzyme marked monoclonal antibody assay.. Serum EGF, E2 and P levels in observed group were significantly higher than those in control group during secretive phase (P < 0.005). The serum EGF level in observed group during prolifrative phase was significantly higher than that of secretive phase (P < 0.05). The serum EGF level increased while uterus enlarged; EGF wasn't related to E2(r = 0.25, P > 0.05) and was related to P (r = 0.71, P < 0.005). The EGF level was the highest when E2 and P both increased.. EGF has the effect on growth of uterine leiomyoma. It is possible that EGF secretion can be stimulated by E2 and the effect of P is stronger than E2.

Topics: Adult; Epidermal Growth Factor; Estradiol; Female; Humans; Leiomyoma; Middle Aged; Progesterone; Uterine Neoplasms

In this work, we report the isolation of a factor from the culture supernatant of confluent fibroblasts from human cervix with the diagnosis of uterine myomatosis. This factor possesses the capacity to inhibit the proliferation of normal fibroblasts. The proliferation inhibitor factor (PIF) was purified from the culture supernatant by precipitation with 80% ammonium sulfate, and by molecular sieve chromatography. Our results indicate that PIF is a protein of 23 kDa, which is highly sensitive to trypsin treatment, and is thermolabile, since temperatures equal to, or above, 60 degrees C eliminate the protein activity in 15 to 20 min. Western blot analyses identified no cross reactions of the purified PIF with TGF-alpha, TNFalpha, IFNgamma, or IL-1beta, suggesting that PIF is a new protein belonging to the group of factors secreted by fibroblasts able to inhibit cellular proliferation.

Topics: Blotting, Western; Cell Count; Chromatography, Gel; Dose-Response Relationship, Drug; Epidermal Growth Factor; Female; Fibroblasts; Gentian Violet; Growth Inhibitors; Humans; Interferon-gamma; Interleukin-1; Leiomyoma; Neoplasm Proteins; Transforming Growth Factor beta; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha; Uterine Neoplasms

Uterine leiomyomas appear during the reproductive years and regress after menopause, indicating the ovarian steroid-dependent growth potential. In order to characterize the molecular mechanism of sex steroidal regulation of leiomyoma growth, we examined whether sex steroids could influence the proliferation of leiomyoma cells. As epidermal growth factor (EGF) has been shown to mediate estrogen action and play a crucial role in regulating leiomyoma growth, we also investigated the effects of sex steroids on EGF and EGF receptor (EGF-R) expression in leiomyoma cells. In cultures of leiomyoma cells, the addition of either estradiol (E(2); 10 ng/ml) or progesterone (P(4); 100 ng/ml) resulted in an increase in proliferating cell nuclear antigen (PCNA) expression in the cells, whereas in cultures of normal myometrial cells, the addition of E(2) augmented PCNA expression in the cells, but P(4) did not. Immunoblot analysis revealed that leiomyoma cells contained immunoreactive EGF and that P(4) treatment resulted in an increase in EGF expression in the cells, whereas E(2) treatment resulted in a lower EGF expression in the cells. By contrast, E(2) treatment augmented EGF-R expression in cultured leiomyoma cells, but P(4) did not. These results indicate that P(4) upregulates the expression of PCNA and EGF in leiomyoma cells, whereas E(2) upregulates the expression of PCNA and EGF-R in those cells. It is, therefore, conceivable that P(4) and E(2) act in combination to stimulate the proliferative potential of leiomyoma cells through the induction of EGF and EGF-R expression. We also found that Bcl-2 protein, an apoptosis-inhibiting gene product, was abundantly expressed in leiomyoma relative to that in normal myometrium and that Bcl-2 protein expression in leiomyoma cells was upregulated by P(4), but downregulated by E(2). It seems, therefore, likely that P(4) may also participate in leiomyoma growth through the induction of Bcl-2 protein in leiomyoma cells. The abundant expression of Bcl-2 protein in leiomyoma cells may be one of the molecular bases for the enhanced growth of a leiomyoma relative to that of normal myometrium in the uterus.

Topics: Adult; Apoptosis; DNA, Neoplasm; Epidermal Growth Factor; ErbB Receptors; Estradiol; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Leiomyoma; Myometrium; Progesterone; Proliferating Cell Nuclear Antigen; Proto-Oncogene Proteins c-bcl-2; Tumor Cells, Cultured; Uterine Neoplasms

Uterine leiomyoma is the most common smooth muscle cell tumor of the myometrium. Estrogen and progesterone (P4) are believed to be physiological regulators of leiomyoma growth. We recently showed that Bcl-2 protein, an apoptosis-inhibiting gene product, was abundantly expressed in leiomyoma relative to its expression in the normal myometrium and that Bcl-2 protein expression in cultured leiomyoma cells was up-regulated by P4, but down-regulated by 17 beta-estradiol (E2). To further characterize the molecular mechanism of sex steroidal regulation of leiomyoma growth, we examined the effect of menstrual phase on proliferating cell nuclear antigen (PCNA) expression in leiomyoma and investigated whether sex steroids could influence PCNA expression in leiomyoma cells cultured under serum-free conditions by immunoblot and immunohistochemical analyses. As epidermal growth factor (EGF) has been shown to mediate estrogen action and to play a crucial role in regulating leiomyoma growth, we also investigated the effects of sex steroids on the expression of EGF and EGF receptor (EGF-R) in cultured leiomyoma cells. The PCNA labeling index in leiomyomas was much greater in the secretory, P4-dominated, phase than in the proliferative phase of the menstrual cycle and was significantly higher than that in the adjacent normal myometrium throughout the menstrual cycle. In monolayer cultures of leiomyoma cells, the addition of either E2 (10 ng/mL) or P4 (100 ng/mL) resulted in an increase in PCNA expression in the cells compared to that in control cultures, whereas in monolayer cultures of myometrial cells, the addition of E2 augmented PCNA expression in the cells, but P4 did not. Immunoblot analysis of proteins extracted from cultured leiomyoma cells revealed that leiomyoma cells contained immunoreactive EGF with a molecular mass of 133 kDa and that the addition of P4 resulted in a remarkable increase in the expression of 133- and 71-kDa immunoreactive EGF in the cells compared to that in control cultures, whereas the addition of E2 resulted in a somewhat lower expression of immunoreactive EGF in the cells. Furthermore, immunocytochemical analysis with a monoclonal antibody to human EGF-R demonstrated that the treatment with E2 augmented EGF-R expression in the cells compared to that in untreated cells, but P4 did not. The concentrations of sex steroids used were within the physiological tissue concentrations found in leiomyomas and myometria. These results indicate that P4

Topics: Adult; Blotting, Western; Epidermal Growth Factor; ErbB Receptors; Estradiol; Female; Humans; Immunohistochemistry; Leiomyoma; Myometrium; Progesterone; Proliferating Cell Nuclear Antigen; Tumor Cells, Cultured; Up-Regulation; Uterine Neoplasms

Hormone receptors and oncoproteins are receiving increased attention as possible prognostic factors in different carcinomas. Few data are available regarding quantification of their levels of expression in gynecologic malignancies.. Epidermal growth factor (EGF) receptor specific binding capacities and affinities were measured by ligand binding assay using [125I]EGF in a competition mode with Accufit software (Lundon Software, Inc., Middlefield, OH). HER-2/neu oncoprotein was extracted from membranes and measured using an enzyme-linked immunosorbent assay. Cathepsin D was measured by an immunoradiometric assay using cytosols for steroid receptor analyses.. EGF receptors in 23 nonmalignant uteri ranged from undetectable to 50 fmol/mg membrane protein (median, 0), with dissociation constant values of 1.2 x 10(-9) M to 8.5 x 10(-10) M, compared with EGF receptors in 76 endometrial cancers that ranged from undetectable to 7674 fmol/mg (median, 52). HER-2/neu oncoprotein ranged from undetectable to 2.9 HER-2/neu units (HNU)/microg protein (median, 0.6) in 41 nonmalignant uteri and from undetectable to 5.8 HNU/microg protein (median, 2.5) in endometrial cancers (n = 53). Cathepsin D ranged from 5 to 32 pmol/mg cytosol protein (median, 11) in 42 nonmalignant uteri and 18 to 144 pmol/mg protein (median, 42) in 29 endometrial cancers.. Determination of the frequency and levels of EGF receptors, HER-2/neu protein, and cathepsin D in uteri with and without cancer and the availability of reference materials developed in our laboratory, will allow evaluation of their prognostic value in cancers of the uterus.

Topics: Cathepsin D; Cell Membrane; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; ErbB Receptors; Female; Humans; Leiomyoma; Prognosis; Radioimmunoassay; Receptor, ErbB-2; Recombinant Proteins; Uterine Neoplasms; Uterus

Uterine leiomyomas (fibroids) are benign, smooth muscle cell (SMC) tumors of the myometrium containing abundant extracellular matrix (ECM). Heparin-binding growth factors present in leiomyoma and normal myometrial fresh tissue were isolated using heparin-affinity fast protein liquid chromatography. Purification of these growth factors was monitored by the stimulation of [3H]thymidine incorporation into BALBc-3T3 cells and myometrial SMC. Western blot analysis confirmed that two consistent peaks of growth factor activity (eluting at 0.5 M NaCl and 1.7 M NaCl) were platelet-derived growth factor (PDGF), 31 kDa, and basic fibroblast growth factor (bFGF), 18 kDa, respectively. Northern blot analysis of leiomyoma and myometrial tissue revealed three RNA transcripts (2.8, 2.3, and 1.9 kb) for PDGF-A chain, one RNA transcript (4.0 kb) for PDGF-B chain, and two RNA transcripts (3.7 and 3.5 kb) for bFGF. RNase protection assay showed elevated expression of the bFGF mRNA transcript in leiomyomas in 3 out of 5 patients. Immunoperoxidase staining of paraffin-embedded tissue showed that PDGF was predominantly intracellular in both vascular and myometrial SMC. Basic FGF, by contrast, was found primarily bound to the ECM of myometrium and fibroids. Leiomyomas showed much stronger staining for bFGF due to the large areas of ECM in these tumors. A third mitogenic peak eluting at 1.1 M NaCl was also seen in both myometrial and leiomyoma tissue. This peak was not definitively identified by Western blotting. However, Northern analysis for heparin binding-epidermal growth factor (HBEGF), which also elutes at 1.1 M NaCl, detected one RNA transcript for HBEGF (2.5 kb) in normal myometrium but little or no expression in the corresponding leiomyoma tissue. Immunoperoxidase staining showed that HBEGF was a cell-membrane-associated protein in both normal myometrial and leiomyoma SMC with more intense staining in normal myometrium. These results show that both leiomyomas and myometrium synthesize a number of heparin-binding growth factors. The enhanced growth of leiomyomas may be due, in part, to the presence of large quantities of bFGF that are stored in the ECM of these tumors. In addition, the level of HBEGF mRNA declines during the transformation of myometrial SMC into leiomyomas.

Topics: Animals; Blotting, Northern; Blotting, Western; Cell Division; Chromatography, Affinity; Epidermal Growth Factor; Female; Heparin-binding EGF-like Growth Factor; Humans; Immunohistochemistry; Intercellular Signaling Peptides and Proteins; Leiomyoma; Mice; Mice, Inbred BALB C; Mitogens; Myometrium; Ribonucleases; RNA, Messenger; Uterine Neoplasms

We studied the estrogen-dependent expression of epidermal growth factor (EGF), transforming growth factor (TGF) alpha, and EGF receptor gene transcripts in human fallopian tubes in vivo and in vitro. Competitive polymerase chain reaction (PCR) was performed on the fallopian tube RNA samples from the postmenopausal women with or without estrogen replacement. Amounts of EGF, TGF alpha, and EGF receptors gene transcripts in the estrogen-treated group (n = 3) were significantly (P < 0.01) more than those in the untreated group (n = 3). Competitive PCR also showed that EGF, TGF alpha, and EGF receptor gene transcripts level in tubal cells were increased by estrogen in vitro: messenger RNA levels of these factors were significantly (P < 0.01, n = 3) increased in cells incubated with 10(-8) M estrogen compared with those in cells without estrogen treatment. We studied whether EGF and/or TGF alpha is involved in the estrogen-induced tubal cell growth in vitro. Estrogen enhanced the [3H]-thymidine incorporation into the cell in dose- and time-dependent manners in culture: estrogen treatment for more than 12 h significantly (P < 0.05) enhanced the [3H]-thymidine incorporation into the cell at 10(-8) M. The estrogen-induced cell growth was observed in association with the increase in EGF, TGF alpha, and EGF receptor messenger RNA levels by estrogen. If the EGF and/or TGF alpha is involved in the cell growth, then the estrogen-induced cell growth should be suppressed by blocking the action of EGF and/or TGF alpha. Therefore, we examined the effects of neutralizing monoclonal antibodies against EGF, TGF alpha, and EGF receptors. Anti-EGF antibody significantly reduced the estrogen-induced increase in [3H]-thymidine incorporation, whereas anti-TGF alpha antibody failed to show the effect. Anti-EGF receptor antibody showed a significant suppressive effect on the estrogen-induced increase in [3H]-thymidine incorporation. Moreover, the growth inhibitory effect by 1 microgram/ml anti-EGF was restored by 10(-8) M EGF but not by TGF alpha even at 10(-6) M. All these data suggest that estrogen induces EGF and TGF alpha/EGF receptors in the human fallopian tube and that EGF but not TGF alpha may be involved in the estrogen-induced human tubal cell growth in vitro.

Topics: Antibodies, Monoclonal; Base Sequence; Catechols; Cells, Cultured; DNA; DNA Primers; Epidermal Growth Factor; Epithelium; ErbB Receptors; Estrogens, Conjugated (USP); Fallopian Tubes; Female; Gene Expression; Growth Inhibitors; Humans; Hysterectomy; Leiomyoma; Middle Aged; Molecular Sequence Data; Nitriles; Polymerase Chain Reaction; Postmenopause; Thymidine; Transforming Growth Factor alpha; Tyrphostins; Uterine Neoplasms

Fibroids (leiomyomata) are the commonest tumours in women, but their aetiology is unknown. Epidermal growth factor (EGF) and transforming growth factor-beta (TGF beta) may be important factors involved in fibroid growth. We examined the mRNA expression of these two growth factors in fibroids and corresponding myometrium from 20 women who underwent hysterectomy because of fibroids. We also examined these factors in samples of fibroids from 9 women who underwent myomectomy after pretreatment with luteinizing hormone-releasing hormone agonists. We found that both factors were expressed in the three types of tissue examined. We also found that there was no difference in the relative abundance of either of the two growth factors between the tissues studied. Despite the lack of difference, we postulate that EGF and TGF beta may be important in fibroid growth because of a possible interaction between the two factors in this tissue.

Topics: Autoradiography; Blotting, Northern; Epidermal Growth Factor; Estrogens; Female; Humans; Leiomyoma; Myometrium; RNA, Messenger; Transforming Growth Factor beta; Uterine Neoplasms

Although there are some reports, including our previous study, indicating the existence and biological action of epidermal growth factor (EGF) in human decidua, the site of its synthesis remains unknown. To clarify the EGF production during decidualization at a molecular level, gene expression of EGF in human endometrium/decidua was examined by Northern blot analysis and in situ hybridization. Northern blot hybridization using 32P-labeled human prepro-EGF complementary DNA was carried out for nonpregnant human endometria from hysterectomized uteri of leiomyoma, decidua from early pregnancy, and in vitro decidualized endometrial cells by medroxyprogesterone acetate (MPA). Although no hybridized band was found in the proliferative and secretory phase endometria, a specific band of 5 kilobases, in agreement with the size of human prepro-EGF messenger ribonucleic acid, was detected in decidua of early pregnancy as well as in in vitro MPA-induced decidual cells. In situ hybridization revealed that prepro-EGF messenger ribonucleic acid was observed in the stromal cells of decidua. These results demonstrate that the EGF gene is expressed in the process of decidualization, suggesting that EGF may play an important role in the decidualization process of human endometrium.

Topics: Blotting, Northern; Decidua; DNA Probes; Endometrium; Epidermal Growth Factor; Female; Gene Expression; Humans; In Situ Hybridization; Leiomyoma; Medroxyprogesterone Acetate; Pregnancy; Protein Precursors; RNA, Messenger; Uterine Neoplasms

Epidermal growth factor (EGF) mRNA was quantified in samples of human myometrium, untreated leiomyomata, and leiomyomata from patients treated with a GnRH analog. Quantitative reverse transcriptase-polymerase chain reaction, using a synthetic internal standard, was applied to determine levels of EGF mRNA. In myometrium from uteri with no leiomyomata, levels of EGF mRNA did not differ between the proliferative and secretory phase of the cycle. Leiomyomata from women who had received no drug therapy had significantly higher amounts of EGF mRNA than myometrium from a normal uterus, but only in the secretory phase of the cycle. In the proliferative phase, leiomyomata did not have different amounts of EGF mRNA compared to normal myometrium. Untreated leiomyomata in the secretory phase of the cycle, but not those in the proliferative phase, had significantly more EGF mRNA than leiomyomata from women who had received treatment with a GnRH analog. These findings suggest that EGF is important in leiomyomata development, but imply that its production is only increased during the secretory phase of the cycle. This challenges the hypothesis that EGF production in leiomyomata is mediated by estrogen and raises the possibility that progesterone may be the more important hormone in fibroid growth.

Topics: Adult; Epidermal Growth Factor; Female; Humans; Leiomyoma; Middle Aged; Myometrium; Polymerase Chain Reaction; RNA, Messenger; Uterine Neoplasms

Immunohistochemical studies were performed using specific antibodies to epidermal growth factor (EGF) and EGF receptor to determine their presence and cellular localization in the human ovary during follicular growth and regression. There was no immunostaining for EGF or EGF receptor in primordial follicles. In the preantral follicle stage, immunostaining for EGF and EGF receptor was observed only in the oocyte. The staining intensity of the oocyte increased as the oocyte reached the preovulatory stage. In the antral follicle stage, immunostaining for EGF and EGF receptor became apparent in the granulosa and theca interna cell layers, without appreciable staining in the surrounding stromal cells. The immunostaining for EGF and EGF receptor in the granulosa cells and theca interna cells persisted in preovulatory follicles and corpus luteum, and intensified in the midluteal phase. The stromal cells surrounding the corpus luteum were negative for EGF and EGF receptor staining. In the regressing corpus luteum, immunostaining for EGF and EGF receptor was present in the peripheral lutein cells adjacent to the central core of scar tissue, but absent in the scar tissue of the central core. Corpus albicans showed no staining for EGF and EGF receptor. By contrast, the stromal cells surrounding the corpus albicans in the cortex region demonstrated intense staining for EGF and EGF receptor, while the stromal cells surrounding the corpus albicans in the medullary region were negative for immunostaining. In the case of atretic follicles, the theca interna cells showed intense staining for EGF and EGF receptor, but immunostaining in the scattered granulosa cells was negligible. This is the first study to demonstrate a remarkable change in the expression of EGF and EGF receptor in the oocyte, granulosa cells, thecal cells, and surrounding stromal cells over the course of follicular growth and regression. The results obtained support EGF participation in oocyte maturation and in follicular growth and atresia. The intense immunostaining for EGF and EGF receptor observed in the theca interna cells in atretic follicles and the stromal cells surrounding corpus albicans in the cortex region raises the possibility of EGF involvement in transformation of thecal cells into stromal cells. Furthermore, the cell type-specific simultaneous expression of EGF and EGF receptor in follicular and stromal compartments in the various stages of follicular development suggests that an autocri

Topics: Adult; Endometriosis; Epidermal Growth Factor; ErbB Receptors; Female; Humans; Hysterectomy; Immunohistochemistry; Leiomyoma; Menstrual Cycle; Ovarian Follicle; Ovariectomy; Ovary; Uterine Cervical Neoplasms

There is a paucity of data concerning the factors involved in the growth of uterine leiomyomata. The expression of mRNAs encoding EGF and the EGF receptor in myometrial and leiomyoma cultures suggest that EGF may be involved in the autocrine/paracrine regulation of human uterine leiomyomata and myometrial growth.

Topics: Amino Acid Sequence; DNA; Epidermal Growth Factor; ErbB Receptors; Female; Humans; Leiomyoma; Molecular Sequence Data; Myometrium; Polymerase Chain Reaction; RNA, Messenger; Transcription, Genetic; Tumor Cells, Cultured; Uterine Neoplasms

The specific binding of epidermal growth factor (EGF), platelet-derived growth factor (PDGF) and insulin were measured in matching cultures of human leiomyoma and myometrial cells, along with the effects of these proteins on DNA and protein syntheses. Scatchard analyses of the binding data revealed that the EGF receptor sites/cell were significantly lower in leiomyoma than myometrial cultures. Two types of PDGF binding were observed when porcine PDGF was used, and one type was seen with human PDGF. By contrast to EGF, more PDGF receptor sites/cell were found in leiomyoma than myometrium but the receptor affinity was higher in the latter. Insulin binding was similar among the myometrial and leiomyoma cells. Protein synthesis was stimulated 3-fold by EGF, PDGF, or insulin in both cell types. DNA synthesis, was higher in myometrial than leiomyoma cells in the basal state and was stimulated by EGF, insulin, or PDGF. A synergistic stimulation (p less than 0.02) of DNA synthesis was observed in both myometrial and leiomyoma cells when EGF was added with insulin. The addition of PDGF with insulin caused only additive stimulation of DNA synthesis. However, the addition of EGF with PDGF caused a synergistic decrease (p less than 0.05) in DNA synthesis by myometrial but no leiomyoma cells. Cultures of human vascular smooth muscle cells obtained from umbilical veins gave results similar to those from myometrium. These findings single out the EGF receptor and EGF, or perhaps an EGF-like growth factor, and to a lesser degree PDGF, as potential regulators of uterine leiomyomata.

Topics: Adult; Analysis of Variance; Cell Division; Culture Techniques; DNA, Neoplasm; Epidermal Growth Factor; ErbB Receptors; Female; Humans; Insulin; Leiomyoma; Middle Aged; Platelet-Derived Growth Factor; Receptor, Insulin; Receptors, Cell Surface; Receptors, Platelet-Derived Growth Factor; Regression Analysis; Tumor Cells, Cultured; Uterine Neoplasms

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.

Topics: Adult; Binding Sites; Epidermal Growth Factor; Female; Humans; Insulin-Like Growth Factor I; Iodine Radioisotopes; Leiomyoma; Middle Aged; Myometrium; Somatomedins; Uterine Neoplasms

The binding of epidermal growth factor (EGF) to human myometrium and leiomyomata was assessed in a group of women rendered hypo-oestrogenic with the LHRH agonist Zoladex (ICI 118630). The results were compared with those obtained with tissues from women with normal cycles. In normal women, the specific binding of radiolabelled [125I] EGF to both myometrial and fibroid homogenates did not vary during the menstrual cycle, but the specific binding of [125I] EGF to fibroid in women treated with LHRH agonist was significantly less than in the untreated group. Since the hypo-oestrogenic state induced by the agonist is associated with a decrease in fibroid size, the results suggest that the effect of oestrogen on fibroid tissue may partly be mediated by EGF.

Topics: Adult; Buserelin; Epidermal Growth Factor; Estradiol; Female; Goserelin; Humans; Leiomyoma; Menstrual Cycle; Myometrium; Uterine Neoplasms