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Histochemical distribution of delta5-3beta- and 17beta-hydroxysteroid dehydrogenases in hamster trophoblast.

The histochemical distribution of delta5-3beta- and 17beta-hydroxysteroid dehydrogenases was demonstrated in hamster trophoblast between Days 8 and 15 of pregnancy. The delta5-3beta-hydroxysteroid dehydrogenase activity in the ectoplacental trophoblast of 8-day embryos was demonstrated by use of delta5-pregnenolone and dehydroepiandrosterone as substrates; between Days 11 and 15, activity was demonstrated in the trophoblastic giant cells of the placenta and in the intra-arterial trophoblast cells when delta5-pregnenolone was the substrate. Between Days 11 and 15, 17beta-hydroxysteroid activity was present in the spongiotrophoblast, labyrinth, placental giant cells and intra-arterial trophoblast cells, as shown by use of testosterone and oestradiol as substrates. Both enzymes were demonstrated in ectopic trophoblast cells, indicating that these activities are autonomous.

17-Hydroxysteroid Dehydrogenases↗

Epitopic heterogeneity of human 3 beta-hydroxysteroid dehydrogenase in villous and extravillous human trophoblast.

A new monoclonal antibody (FDO26G) is described which was raised against purified human 3 beta-hydroxysteroid dehydrogenase type I (3 beta-HSD type I). FDO26G reacted strongly with villous syncytiotrophoblast, weakly with some trophoblast cells in chorion laeve, and not at all with extravillous trophoblast in cytotrophoblast cell islands and decidual trophoblast. All these types of trophoblast reacted strongly with monoclonal antibody FDO161G, which has previously been shown to react with 3 beta-HSD type I and, like FDO26G, reacts strongly with adrenal cells. Mapping experiments using a combination of lacZ fusion polypeptides and synthetic peptides located the FDO26G epitope to residues 354-366 at the C-terminal end of the molecule, a sequence that is identical in the type I and type II forms of the enzyme. The epitope contains a consensus for a casein kinase-II site with serine 359 as the candidate phosphorylation site. This suggested that the lack of reactivity of FDO26G to 3 beta-HSD in extravillous trophoblast might be due to phosphorylation at serine 359. Peptide 354-366 was synthesized with phosphoserine at residue 359 and its binding to FDO26G was compared with that of the unphosphorylated peptide. FDO26G bound the phosphopeptide at least as strongly as the unphosphorylated peptide. It is concluded that the lack of staining of extravillous trophoblast by FDO26G is due to the presence of a different sequence at residues 354-366 and that a hitherto unidentified third isoform of human 3 beta-HSD is expressed in these cells.

3-Hydroxysteroid Dehydrogenases↗

Immunohistochemistry in the diagnosis of the gestational trophoblastic disease.

OBJECTIVE: The gestational trophoblastic disease summarizes all types of hydatidiform moles, placental site trophoblastic tumor and choriocarcinoma. It is of clinical relevance to distinguish between complete hydatidiform mole and partial hydatidiform mole to predict prognosis of recurrency of molar pregnancy and the risk of the development of malign and metastatic gestational trophoblastic disease. Differential diagnosis of choriocarcinoma versus placental site trophoblastic tumor, carcinoma or sarcoma with low differentiation can cause problems in borderline-cases. The present study investigates the value of immunohistochemistry in the diagnosis of gestational trophoblastic disease. METHOD: Nine cases of patients with complete hydatidiform mole, 20 cases of partial hydatidiform mole and seven cases of choriocarcinoma were analyzed for the immunohistochemical reaction with antibodies against human choriogonadotropin (hCG), human placental lactogen (hPL). placental alkaline phosphatase (PLAP), cytokeratine and vimentin. RESULTS: Complete hydatidiform mole shows strong expression of hCG and weak expression of PLAP. Weak hCG and strong PLAP expression is found in partial hydatidiform mole. Choriocarcinoma presents strong expression of hCG and weak expression of hPL and PLAP. All tissues show positive reaction with anticytokeratine and negative reaction with anti-vimentin. CONCLUSION: Our study proves immunohistochemistry as useful tool for differential diagnosis in borderline cases of gestational trophoblastic disease.

Alkaline Phosphatase↗

Human first-trimester trophoblast cells recruit CD56brightCD16- NK cells into decidua by way of expressing and secreting of CXCL12/stromal cell-derived factor 1.

More than 70% of decidual lymphocytes are NK cells characterized by CD56(bright)CD16(-) phenotype, but the mechanisms by which these NK cells are recruited in the decidua are still almost unrevealed. In this study, we first analyzed the transcription of 18 chemokine receptors in the first-trimester decidual CD56(bright)CD16(-) NK cells. Among these receptors, CXCR4 and CXCR3 were found highly transcribed, and the expression of CXCR4 was verified in most of the decidual CD56(bright)CD16(-) NK cells by flow cytometry. The first-trimester human trophoblasts were found expressing CXCL12/stromal cell-derived factor 1, the specific ligand of CXCR4, by way of in situ hybridization and immunohistochemistry. The primary cultured trophoblast cells were also found to secrete stromal cell-derived factor 1alpha spontaneously, and its concentration was 384.6 +/- 90.7 pg/ml after the trophoblast cells had been cultured for 60 h. All of the ligands for CXCR3 were below the minimal detectable concentration when trophoblast cells were cultured for up to 48 h. Both recombinant human SDF-1alpha and supernatants of the cultured trophoblast cells exhibited chemotactic activity on decidual CD56(bright)CD16(-) NK cells. Our findings suggest that human first-trimester trophoblast cells produce CXCL12, which in turn chemoattracts decidual CD56(bright)CD16(-) NK cells. This activity could contribute to the recruitment mechanism of decidual lymphocytes, especially CD56(bright)CD16(-) NK cells, in decidua, and may be used at a local level to modulate the immune milieu at the materno-fetal interface.

Base Sequence↗

Trophoblast contact deactivates human neutrophils.

Trophoblasts are fetal epithelial cells that form an interface between mother and offspring. To evaluate their anti-inflammatory capacity, we tested the hypothesis that trophoblasts deactivate neutrophils using single-cell assays. Several biophysical (Ca2+ and NAD(P)H oscillation frequency) and physiological (oxidant production) markers of activated neutrophils revert to a nonactivated phenotype as activated cells make contact with trophoblasts. Indistinguishable results were obtained using syncytiotrophoblasts and in experiments using trophoblasts and neutrophils from the same mother to recapitulate the semiallogeneic system. These changes suggest reduced hexose monophosphate shunt (HMS) activity. We discovered that two metabolic regulatory points, glucose transport and HMS enzyme trafficking, are affected by trophoblasts. This restriction in HMS activity deactivates neutrophils, thereby limiting oxidative DNA damage within trophoblasts.

Calcium Signaling↗

Sonohysterography for the diagnosis of residual trophoblastic tissue.

OBJECTIVE: To assess the efficacy, safety, and associated complications of sonohysterography for the diagnosis of residual trophoblastic tissue. METHODS: We conducted a prospective study of 23 consecutive patients admitted to our ultrasonography unit with clinical and ultrasonographic signs of retained intrauterine tissue. RESULTS: Twelve patients had hydrosonographic features suggestive of residual trophoblastic tissue (i.e., an intrauterine lesion not detachable from the uterine wall after instillation of saline), whereas in 11 cases the hydrosonographic findings were negative for retained tissue. Blood flow was detected within abnormal intrauterine masses in 4 of 12 patients with trophoblastic tissue, whereas it was not detected in any patient without retained tissue (P = .093). No complications were encountered during the procedure or the postprocedure period. None of the patients had anesthetic complications, perforation of the uterus, fluid overload, or any other surgical complication. All 12 patients underwent hysteroscopic removal of the suspected residual trophoblastic tissue, and histologic confirmation of residual trophoblastic tissue was obtained in all cases. CONCLUSIONS: Sonohysterography for detection and diagnosis of residual trophoblastic tissue is an accurate and safe procedure. Further studies comparing the efficacy of sonohysterography with that of diagnostic hysteroscopy are warranted.

Abortion, Induced↗

Expression of the human P2Y6 nucleotide receptor in normal placenta and gestational trophoblastic disease.

The P2Y family of purinergic receptors are members of the G-protein-coupled receptor superfamily. The P2Y6 subtype is expressed at particularly high levels in the placenta, suggesting that P2Y6 plays an important role in placental function. However, the cellular localization of P2Y6 within the placenta is unknown. This study examined the expression of P2Y6 in first-trimester and full-term placental tissues, as well as examples of gestational trophoblastic disease, by Northern blot analysis and in situ hybridization. P2Y6 message was present at similar levels in first-trimester and full-term placenta, and in situ hybridization revealed that message was most abundant in the cytotrophoblast of the villi and chorionic plate at both gestational stages. The syncytiotrophoblast harbored lower levels of P2Y6 in first-trimester placenta, and by full-term, the syncytiotrophoblast only focally expressed P2Y6 transcripts. Neither the intermediate trophoblast nor nontrophoblastic elements of the placenta expressed P2Y6. Molar disease expressed P2Y6 in the villous trophoblast but not in the proliferative intermediate trophoblast, recapitulating the pattern of first-trimester placenta. Neither choriocarcinoma nor the choriocarcinoma cell lines JEG-3 and JAr expressed P2Y6 transcript. These findings reveal that P2Y6 mRNA production is highly characteristic of the epithelial-like cytotrophoblast and syncytiotrophoblast, whereas expression is absent in the mesenchymal-like intermediate trophoblast. Thus, P2Y6 may play an important role in trophoblastic development, differentiation, and neoplasia.

Blotting, Northern↗

A study of early pregnancy factor activity in the sera of women with trophoblastic tumor.

PROBLEM: To detect whether or not the early pregnancy factor (EPF)-like activity, or chaperonin 10, could be in the sera of patients with trophoblastic tumor in order to find another more efficient means to diagnose this kind of tumor. METHOD OF STUDY: The rosette inhibition assay was used to detect EPF-like activity in 216 sera, collected from patients with gestational trophoblastic tumor, including 47 sera of patients with choriocarcinoma, 68 sera of patients bearing invasive mole, and 101 sera of patients with vesicular mole. RESULTS: The accuracy of diagnosing malignant trophoblastic tumor by detecting EPF-like activity is 91.3% (105/115), with a false positive rate of 14.58% and a false negative rate of 4.8% by this method. Furthermore, the rosette inhibition titer (RIT) values have significant difference (P < 0.001) between the sera in patients with malignant trophoblastic tumor before treatment and those after treatment. CONCLUSIONS: This study demonstrated that diagnosis of malignant trophoblastic tumor could be made with an accuracy of 91.3% by detecting EPF-like activity and that EPF-like activity could be used as an indicator to distinguish benign from malignant trophoblastic tumor.

Adult↗

[Fertility after chemotherapy of gestational trophoblastic disease].

OBJECTIVE: To evaluate subsequent pregnancy experience in patients following chemotherapy for malignant form of gestational trophoblastic disease. DESIGN: Retrospective clinical study. SETTING: Slovak Center of Trophoblastic Disease, Bratislava, Slovak Republic. METHODS: There were evaluated subsequent pregnancy experiences in 38 patients after chemotherapy for malignant form of gestational trophoblastic disease registered in Slovak center of gestational trophoblastic disease. Histological and cytogenetical analysis of all placentas after deliveries and material from curettage specimens after miscarriages, abortions and ectopic pregnancies were performed. RESULTS: 11 women conceived following successful chemotherapy of gestational trophoblastic disease became pregnant a total 19 times. Out of them there were 9 full-term deliveries, 2 spontaneous abortion, 1 ended in ectopic pregnancy and 7 pregnancies were terminated in therapeutic abortion. Cytogenetical analysis was successful in 7 to 10 reproductive losses with normal karyotype in all analysed cases. CONCLUSION: Patients after successful chemotherapy of gestational trophoblastic disease have a normal reproductive outcome.

Antineoplastic Agents↗

Utility of commonly used commercial human chorionic gonadotropin immunoassays in the diagnosis and management of trophoblastic diseases.

BACKGROUND: Patients with trophoblastic diseases produce ordinary and irregular forms of human chorionic gonadotropin (hCG; e.g., nicked hCG, hCG missing the beta-subunit C-terminal segment, hyperglycosylated hCG, and free beta subunit) that are recognized to differing extents by automated immunometric hCG (or hCG beta) assays. This has led to low or false-negative results and misdiagnosis of persistent disease. False-positive hCG immunoreactivity has also been detected, leading to needless therapy for trophoblastic diseases. Here we compare seven commonly used hCG assays. METHODS: Standards for five irregular forms hCG produced in trophoblastic diseases, serum samples from 59 patients with confirmed trophoblastic diseases, and serum samples from 12 women with previous false-positive hCG results (primarily in the Abbott AxSYM assay) were blindly tested by commercial laboratories in the Beckman Access hCG beta, the Abbott AxSYM hCG beta, the Chiron ACS:180 hCG beta, the Baxter Stratus hCG test, the DPC Immulite hCG test, the Serono MAIAclone hCG beta tests, and in the hCG beta RIA. RESULTS: Only the RIA and the DPC appropriately detected the five irregular hCG standards. Only the Beckman, DPC, and Abbott assays gave results similar to the RIA in the patients with confirmed trophoblastic diseases (values within 25% of RIA in 49, 49, and 54 of 59 patients, respectively). For samples that were previously found to produce false-positive hCG results, no false-positive results were detected with the DPC and Chiron tests (5 samples, median <2 IU/L), but up to one-third of samples were false positive (>10 IU/L) in the Beckman (1 of 5), Serono (2 of 9), and Baxter assays (1 of 5), and the hCG beta RIA (3 of 9; median for all assays, <5 IU/L). These samples, which produced false-positive results earlier in the Abbott AxSYM assay, continued to produce high values upon reassessment (median, 81 IU/L). CONCLUSIONS: Of six frequently used hCG immunometric assays, only the DPC detected the five irregular forms of beta hCG, agreed with the RIA, and avoided false-positive results in the samples tested. This assay, and similarly designed assays not tested here, seem appropriate for hCG testing in the diagnosis and management of trophoblastic diseases.

Choriocarcinoma↗

[Clinical study of four cases with malignant gestation trophoblastic tumor after mifepristone abortion].

OBJECTIVE: To describe the clinical characteristics of malignant gestational trophoblastic tumor after medical abortion used by mifepristone combined with misoprostol and its diagnosis and differential diagnosis from incomplete abortion. METHODS: Four cases with malignant gestational trophoblast tumor after medical abortion were presented focusing on the clinical manifestation and the methods of diagnosis and differential diagnosis. RESULTS: Irregular vaginal bleeding and abnormal high level of beta-human chorionic gonadotropin (hCG) in plasma were the common manifestation of the gestational trophoblast tumour and incomplete abortion after medical abortion. However, beta-hCG of the former after curettage was still higher by dynamic monitoring. Malignant gestational trophoblast tumor showed rich blood flow signal and low blood flow resistance index (RI, RI < 0.5) in uterus in color doppler echography, digital subtraction angiography (DSA) with abnormal enlargement of the arteria of uterine, arteriovenous fistula beside the uterine were the main characteristics of malignant gestational trophoblast tumour. CONCLUSIONS: Pay attention to the early stage malignant gestational trophoblast tumour among patients with abnormal vaginal bleeding after medical abortion. beta-hCG and DSA were the most effective methods to diagnose and differentially diagnose choriocarcinoma from the incomplete abortion among the patients with abnormal vaginal bleeding after medical abortion.

Abortifacient Agents, Steroidal↗

Infantile hemangiomas: speculation on placental trophoblastic origin.

BACKGROUND: The unique immunobiology of the placental trophoblast and the increased incidence of hemangiomas in infants born after chorionic villus sampling suggest that an immunologically regulated ectopic focus of trophoblasts could be the cell of origin for proliferative infantile hemangiomas. OBJECTIVE: To compare tissue from infantile hemangiomas with that of other vascular lesions for the presence of selected placental trophoblast-specific cellular markers. DESIGN AND PATIENTS: Twelve tissue specimens taken from infantile hemangiomas on patients aged 5 days to 2 years were retrospectively confirmed clinically and histologically. Negative controls were similarly confirmed, including 6 pyogenic granulomas and 4 vascular-lymphatic malformations. These tissues were used for immunohistochemical analysis of selected trophoblastic markers including human placental lactogen, placental alkaline phosphatase, and cytokeratins 7, 8, and 17. SETTING: Tissue submitted from patients seen at Saint Louis University Department of Dermatology and Cardinal Glennon Children's Hospital in St Louis, Mo, between January 1, 1997, and October 31, 1999. MAIN OUTCOME MEASURE: Differential staining for trophoblastic markers in infantile hemangiomas compared with control tissues. RESULTS: The 12 infantile hemangiomas were uniformly negative for all markers tested. Control tissues were also negative for these markers. Four of the 5 histochemical markers did recognize specific nonvascular, cutaneous elements: placental alkaline phosphatase stained smooth and striated muscle, cytokeratins 7 and 8 stained eccrine glands, and cytokeratin 17 stained pilosebaceous units. CONCLUSIONS: Our results do not support the placental trophoblast as the cell of origin for infantile hemangiomas, but we hope our observations and speculation will stimulate further study of this hypothesis.

Alkaline Phosphatase↗

[Expression of epidermal growth factor receptor in human endometrium, decidua and trophoblast cells].

OBJECTIVE: To study the expression of epidermal growth factor receptor (EGFR) expression in human normal endometrium, decidua and trophoblast cells of early pregnancy, and to explore the effect of EGFR on cell proliferation and development. METHODS: To localize the EGFR expression in glandular and stromal cells of normal endometrium, decidua with immunohistochemical method (ABC), and to analyse the related quantity of EGFR mRNA by RT-PCR method. RESULTS: There were EGFR expression in both normal endometrium and decidua. EGFR protein located on cell membrane, nuclear membrane and in the cytoplasm as well, and located in the neuclear of glandular and stromal cells in decidua. The expression of EGFR in glandular cells was significantly stronger than that in the stroma (P < 0.05), while there was no difference of EGFR expression in the glands during the menstrual cycle (P > 0.05). However, the expression of EGFR in the glands of decidua was significantly higher than that in the proliferative and secretory endometrium (P < 0.05). The intensity of staining in stromal cells listed in order from the strongest was early decidua > secretory endometrium > proliferative endometrium, the differences were significant (P < 0.05). The related quantity of EGFR mRNA expression in proliferative, secretory endometrium, decidua and trophoblasts were: 0.531 +/- 0.061, 0.594 +/- 0.121, 0.732 +/- 0.032, 0.828 +/- 0.034, respectively. There was no difference during the menstrual cycle, but that in the decidua and trophoblasts were significantly higher than that in the proliferative and secretory endometrium (P < 0.05), and that in the trophoblasts was the highest. CONCLUSION: EGFR expression was present in the proliferative and secretory endometrium, decidua and trophoblasts of early pregnancy, and it was significantly higher in the decidua and trophoblasts than that during the menstrual cycle.

Adult↗

Negotiating a staging and risk factor scoring system for gestational trophoblastic neoplasia. A progress report.

OBJECTIVE: To discuss the current problems in establishing a unified staging and risk factor scoring system for trophoblastic disease under the auspices of the International Federation of Gynecology and Obstetrics (FIGO). STUDY DESIGN: To present a history of the classifications of trophoblastic disease and compare these with the proposals presented to FIGO by the International Society for the Study of Trophoblastic Disease, the International Gynecologic Cancer Society and the Society of Gynecologic Oncologists. RESULTS: The situation concerning the proposed 2000 FIGO trophoblast classification was resolved by FIGO's acceptance of the recommendations outlined in this paper. CONCLUSION: There is a scientific and clinical need for FIGO to adopt the recommendations of the majority of physicians with expertise in treating trophoblastic disease for a rational classification of trophoblastic disease. It is gratifying that FIGO has done so.

Consensus↗

Multiclefted nuclei. A helpful feature for identification of intermediate trophoblastic cells in uterine curetting specimens.

Intermediate trophoblast is a distinct form of trophoblast, the presence of which in uterine curettings is considered a reliable indicator of intrauterine pregnancy even in the absence of chorionic villi. However, the appearance of intermediate trophoblastic cells have not been described in sufficiently specific terms to permit their reliable identification, and distinction from decidual cells can be difficult. We have noticed for some time that the intermediate trophoblastic cells often show multiple deep clefts in the nuclei, and the present study was performed to address the issue of whether this nuclear feature is reliable for their identification. We reviewed 242 uterine curettings of intrauterine pregnancy, documented by the presence of chorionic villi, and were able to find a distinct population of cells with large, hyperchromatic, multiclefted nuclei scattered in the decidua in 88% of the cases. In most instances, these cells produced a characteristic variegated pattern that was readily recognizable at low magnification. Positive immunostaining for cytokeratin (CAM 5.2) in these isolated cells within the decidua confirmed their trophoblastic nature. In contrast, multiclefted nuclei were absent in the 51 negative control cases, which included decidualized endocervical polyps (40 cases), uterine curettings from patients with tubal pregnancy (10 cases), and endometriosis with decidual change (one case). We conclude that intermediate trophoblastic cells can usually be reliably identified in curettings of intrauterine pregnancy by their characteristic nuclear multiclefting.

Cell Nucleus↗

Diagnostic aspects of hydatidiform mole with persistence of polymorphic trophoblastic hyperplasia.

UNLABELLED: The persistence of polymorphic trophoblastic hyperplasia in a hydatidiform mole is an extremely rare condition. Its early recognition is essential since such cases can transform into invasive types of tumors. MATERIALS AND METHODS: The biopsies were routinely processed in paraffin, embedded and stained with HE. Immunohistochemical staining reactions were performed with the following monoclonal antibodies for hydatidiform mole: beta-hCG, HPL, MIB1, CK18, HER-2/neu, p53 and carbohydrate antibodies, Thomsen-Friedenreich antigen, Glycodelin A, Mucl and Mucl-cor. RESULTS: Large villi and hydatidiform villi with wide-ranged syncyctio- and cytotrophoblasts were seen. Intervillous proliferating trophoblasts showed cell- and nuclear polymorphy with a wall invasion of the myometrium. The immunohistochemistry exhibited strong positivity for the membrane-associated HER-2/neu and for the beta-hCG in syncytiotrophoblast and in multinuclear giant cells of intervillus trophoblasts. A weakly positive reaction with hPL was seen in most cells of the trophoblasts. The rest of the immunohistochemistry served as a diagnostic support. CONCLUSION: A complete hydatidiform mole with hyperplasia and proliferation of polymorphic trophoblasts presents a high risk of developing a persistent (eventually metastatic) trophoblastic disorder and, in up to 15% of the cases, an invasive mole. In 2.5% of the cases it can transform into a choriocarcinoma.

Adult↗

Effects of phytoestrogens on the trophoblast tumour cell lines BeWo and Jeg3.

INTRODUCTION: Phytoestrogens are a diverse group of nonsteroidal plant compounds that occur naturally in many plants. Because they possess a ring system similar to estrogens they are able to bind to estrogen receptors in humans. With this study we tested the effects of the phytoestrogens genistein and daidzein in cell proliferation and the production of progesterone and hCG in trophoblast tumour cells of the cell lines BeWo and Jeg3. MATERIALS AND METHODS: The phytoestrogens genistein and daidzein were incubated in different concentrations with trophoblast tumour cells. Untreated cells were used as controls. At designated times, aliquots were removed and tested for progesterone and hCG. In addition we tested the effects of phytoestrogens on cell proliferation. Different concentrations of genistein and daidzein were cultivated with trophoblast tumour cells. After designated times, 1 microCi thymidin-(methyl-3H) was added. Methyl-3H thymidin incorporation was tested and compared to incorporation results of untreated cells. RESULTS: With this study we could show that the production of the steroid hormone progesterone and the protein hormone hCG is influenced by the phytoestrogens genistein and daidzein in trophoblast tumour cells of the cell lines BeWo and Jeg3. We found a correlation between the effects on the proliferation and the production of progesterone and hCG at high concentrations of genistein and daidzein in the cell lines tested. With low concentrations of genistein and daidzein we observed a stimulation of the production of hCG and a weak inhibition of proliferation in both cell lines BeWo and Jeg3. DISCUSSION: The results obtained with this study suggest that only high doses of phytoestrogens (> 1 mumol/ml) can reduce the proliferation of trophoblast tumour cells significantly. Low doses of phytoestrogens induced a higher hCG production in both cell lines tested. Although high hCG production did not lead to a higher proliferation rate of the tumour cells tested, hCG is able to induce neovascularisation in tumour cells. In summary, with this in vitro study we showed that high doses of phytoestrogens inhibit proliferation and progesterone production in trophoblast tumour cells. High doses of phytoestrogens could be useful candidates for special diet programs for prevention and surgery for patients with this type of disease. In addition we found a useful cell culture model for the testing of new types of phytoestrogens.

Cell Division↗

hCG in trophoblast tumour cells of the cell line Jeg3 and hCG isolated from amniotic fluid and serum of pregnant women carry oligosaccharides of the sialyl Lewis X and sialyl Lewis a type.

Human chorionic gonadotropin (hCG) is a placental hormone and marker for the differentiation process of cytotrophoblast cells to syncytial trophoblasts. It is secreted into the maternal circulation and urine. The glycoprotein hCG is also found in the serum and urine of patients with malignant trophoblastic diseases. For this study hCG was purified from pooled human mid-trimester amniotic fluid, serum and urine of pregnant women and from supernatants of trophoblast tumour cell cultures Jeg3 and BeWo by immunoadsorption chromatography with specific hCG-antibodies. The purity and identity of the isolated hCG were checked by Western blot analysis. Sialylated oligosaccharides of the Lewis type of isolated hCGs were analysed by dot blot and ELISA technique with monoclonal antibodies specific for sialyl Lewis x and sialyl Lewis a-carbohydrate epitopes. BeWo and Jeg 3 cells were additionally investigated by immunofluorescence. HCG isolated from the serum and amnion of pregnant women bear both sialyl Lewis x (sLex)-antigen and sialyl Lewis a (sLea)-antigen. The same result was observed for hCG isolated from the supernatants of the trophoblast tumour cell line Jeg3. On the contrary, hCG isolated from the supernatants of the trophoblast tumour cell line BeWo showed only a very weak expression of these antigens. HCG isolated from the urine of pregnant women was negative for both sialyl Lewis antigens. The results are discussed with respect to a possible relationship to selectin- mediated cell adhesion processes. This could play a role in the prevention of leukocyte adhesion on the foetal syncytiotrophoblast. Glycosylation of hCG in trophoblast tumour cells, on the other hand, could increase the metastatic activity of these cells.

Amniotic Fluid↗