Human Chorionic Gonadotropin Production Research Articles

GDF-11 downregulates placental human chorionic gonadotropin expression by activating SMAD2/3 signaling

BackgroundThe production of human chorionic gonadotropin (hCG) by the placental trophoblast cells is essential for maintaining a normal pregnancy. Aberrant hCG levels are associated with reproductive disorders. The protein of hCG is a dimer consisting of an α subunit and a β subunit. The β subunit is encoded by the CGB gene and is unique to hCG. Growth differentiation factor-11 (GDF-11), a member of the transforming growth factor-β (TGF-β) superfamily, is expressed in the human placenta and can stimulate trophoblast cell invasion. However, whether the expression of CGB and the production of hCG are regulated by GDF-11 remains undetermined.MethodsTwo human choriocarcinoma cell lines, BeWo and JEG-3, and primary cultures of human cytotrophoblast (CTB) cells were used as experimental models. The effects of GDF-11 on CGB expression and hCG production, as well as the underlying mechanisms, were explored by a series of in vitro experiments.ResultsOur results show that treatment of GDF-11 downregulates the expression of CGB and the production of hCG in both BeWo and JEG-3 cells as well as in primary CTB cells. Using a pharmacological inhibitor and siRNA-mediated approach, we reveal that both ALK4 and ALK5 are required for the GDF-11-induced downregulation of CGB expression. In addition, treatment of GDF-11 activates SMAD2/3 but not SMAD1/5/8 signaling pathways. Moreover, both SMAD2 and SMAD3 are involved in the GDF-11-downregulated CGB expression. ELISA results show that the GDF-11-suppressed hCG production requires the ALK4/5-mediated activation of SMAD2/3 signaling pathways.ConclusionsThis study not only discovers the biological function of GDF-11 in the human placenta but also provides important insights into the regulation of the expression of hCG.EYYrMgGK1oNKot-fDMBUfiVideo

Abstract #1404395: Metastatic Choriocarcinoma: A Rare Cause of Thyrotoxicosis

Although Graves’ disease continues to be considered the most frequent cause of clinical hyperthyroidism in pregnancy, the disproportionately high production of human chorionic gonadotropin (HCG) as occurs in gestational trophoblastic disease (TSG), is another possible cause that should be considered when evaluating thyrotoxicosis in pregnancy. A 19-year-old female with unremarkable past medical history who was admitted for nausea, vomiting and abdominal pain for 2 weeks. Her last period was 4 weeks prior to her presentation and patient denied any vaginal bleeding. Her period had been irregular since her last miscarriage a year earlier. She denied being sick recently, and her last flulike illness was 4 months ago. Denied using any medication at home including herbal medication or OCPs. Review of systems was significant for fatigue, heat intolerance, palpitation, tremor, loss of appetite and unintentional weight loss over the last week. She denied diarrhea or anxiety. On presentation, Patient was hypotensive 90s/40s mmHg and tachycardic 140s bmp. EKG showed sinus tachycardia. Initial work-up was significant for HB 6.2, WBC 11.6 and B-hCG 9678 IU/ml. Liver enzymes were elevated. Transvaginal ultrasound showed empty uterus. Pan CT showed hemoperitonium and liver, lung and kidney lesions concerning for metastatic choriocarcinoma which was confirmed with positive liver biopsy. Work-up also showed suppressed TSH 0.01uIU/ml, high free T4 2.54ng/dl, and high free T3 5.1ng/dl, absent TSI, normal Tg level with negative Tg antibodies. There was no baseline TSH to compare. BHCG with dilution was 5,430,055IU/ml. Thyroid ultrasound was normal. Patient was diagnosed with metastatic choriocarcinoma and treated with 3 cycles of cisplatin/etoposide chemotherapy. Patient did not receive any antithyroid medication due to elevated liver enzymes and mild symptoms. B-HBG following the treatment was 3mIU/ml, TSH was 1.5 uIU/ml and FT4 was 1.1ng/dl. Hyperthyroidism has been reported in many patients with GTD. The beta subunit of β-hCG is structurally similar to TSH, allowing it to bind to the TSH receptor on thyroid follicular cells. It has been reported that β-hCG levels of greater than 200,000 m[IU]/ml sustained for several weeks are required to induce clinical hyperthyroidism. It is estimated that for every 10,000 m[lU]/ml increase in serum hCG, TSH decreases by 0.1 mlU/ml and free T4 increases by 0.1 ng/dL. No clear risk factors for hyperthyroidism in GTD were identified. Presentations can range from subclinical hyperthyroidism to thyrotoxicosis and thyroid storm. This case was challenging because the initial B-HCG was not high enough to cause thyrotoxicosis but delusional B-HCG confirmed our suspicion. While hyperthyroidism can be definitively managed with surgical evacuation of the uterus in GTD, hyperthyroidism secondary to metastatic choriocarcinoma could be successfully treated with chemotherapy with or without antithyroid medication. The increase in TSH usually parallel the reduction in BHCG.

Characterization of Three-Dimensional Trophoblast Spheroids: An Alternative Model to Study the Physiological Properties of the Placental Unit.

It was postulated that 3D cell culture models more accurately reflect the complex tissue physiology and morphology in comparison to 2D cell monolayers. Currently, there is a shortage of well-characterized and easily maintainable high-throughput experimental models of the human placenta. Here, we characterized three different 3D cultures (e.g., spheroids) derived from trophoblast cell lines and studied their functionality in comparison to primary fetal trophoblasts and placental tissue. The spheroid growth rates of JEG3, BeWo and HTR8/SVneo cell lines were similar among each other and were significantly larger in comparison to primary trophoblast spheroids. All spheroids exhibited migratory properties and shortest distances were registered for JEG3 spheroids. Even though all spheroids displayed invasive capabilities, only the invasive features of HTR8/SVneo spheroids resulted in specific branching. This was in agreement with the invasive properties of the spheroids obtained from primary trophoblasts. Human chorionic gonadotropin production was highest in JEG3 spheroids and only increased when stimulated with cAMP and forskolin in BeWo, but not HTR8/SVneo spheroids. The gene expression analysis confirmed that 3D trophoblast cell cultures and especially HTR8/SVneo spheroids showed considerable similarities with the gene expression profile of primary placental tissue. This study offers a broad characterization of 3D trophoblast spheroids that, in turn, can help in selecting the best model depending on the scientific question that needs to be answered.

Amenorrhea and pituitary human chorionic gonadotrophin production in a 38-year-old presenting as pregnancy of unknown location: case report and review of literature

Background: Amenorrhea and extraplacental production of serum human chorionic gonadotropin (hCG), particularly in young women, can mimic a pregnancy of unknown location. Elevated serum hCG in the absence of pregnancy can pose a diagnostic dilemma and has led to potentially harmful and unwarranted interventions including chemotherapeutic agents like methotrexate or have led to delay in necessary medical interventions in women. We report a case to demonstrate that amenorrhea and extraplacental human chorionic gonadotropin (hCG) production in young women can mimic a pregnancy of unknown location. Furthermore, we performed a critical review of literature on pituitary hCG production. Case: A 38-year-old woman with a diagnosis of Silver-Russell syndrome, a unicornuate uterus, history of right oophorectomy for a benign serous cystadenoma and a desire for pregnancy presenting with a provisional diagnosis of pregnancy of unknown location.After performing a thorough review of history, physical examination, ultrasound exams, and a review of hormone analysis [including hCG, Tumor markers, Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), Anti-Mullerian Hormone (AMH), Estradiol (E2) levels], we confirmed the diagnosis of premature ovarian insufficiency and pituitary hCG production. Conclusions: In women, serum levels of hCG may increase with age, and are not always an indicator of pregnancy. Therefore, it is imperative to interpret false-positive test results and rule out the extraplacental production of hCG. This will help prevent unnecessary surgical procedures and treatment, including chemotherapy.

MATERNAL PHYSIOLOGY AND BLASTOCYST MORPHOLOGICAL GRADE CORRELATE WITH AN INHERENT DIFFERENCE IN HUMAN PERI-IMPLANTATION DEVELOPMENTAL POTENTIAL

Our group has been using an extended embryo culture (EEC) system to culture human embryos for up to an additional 7 d to study peri-implantation over the past three years. Since similar EEC parameters were measured for these projects, we performed a retrospective analysis of 653 human embryos to determine what maternal factors correlate with peri-implantation developmental outcomes. Frozen day five or day six human blastocysts donated for research (WIRB study no. 1179872) were warmed, dezonated, and cultured in vitro up to EEC day seven following an established protocol (Deglincerti et al., Nature 2016). The trophectoderm outgrowth area, total cell number, epiblast cell number, and human chorionic gonadotropin (hCG) production in spent media were measured during EEC. Patient information including maternal BMI, age, infertility diagnosis, FSH, LH, FSH: LH ratio, estradiol, and anti-müllerian hormone (AMH) levels on menstrual cycle day 3, and an antral follicle count (AFC) were used in our analysis. Additionally, blastocysts with the highest morphological grade that typically yield live birth rates above 65% (Good Quality) were compared with those of lower morphological grade that typically yield less than 55% live birth rate (Fair Quality), based on our historical data. We then used a spearman correlation coefficient to assess the strength of the association between the aforementioned maternal factors and extended culture parameters. A chi-squared analysis was used to compare binomial outcomes with categorical predictors. T tests or ANOVA’s were used for comparing two or multiple continuous variables from categorical predictors respectively. Several maternal physiological influences during oocyte development (age, BMI, and FSH: LH, AMH, AFC) were significantly correlated with extended culture outcomes (p<0.05). Embryos with high morphological grade had significantly increased outgrowth areas compared to those with low morphological grade (Good Quality: 0.17 ± 0.02; Fair Quality: 0.12 ± 0.02, p<0.05). Maternal age was correlated with hindered EEC measurements in both the non-donor fertility patients as well as in healthy donors. When comparing embryos with the highest morphological grade only, advanced maternal age (35+ years old) still contributed to decreased hCG production (p<0.05) and hindered epiblast formation compared to those from younger patients (<35 years old, p<0.01). In conclusion, we show that human EEC may be valuable in predicting post-transfer developmental potential. Additionally, maternal physiology during oocyte development including age, BMI, and FSH: LH, AFC, and AMH significantly correlates with the peri-implantation developmental potential of human embryos.

Maternal Obesity Influences Placental Nutrient Transport, Inflammatory Status, and Morphology in Human Term Placenta.

Maternal obesity has a significant impact on placental development. However, this impact on the placenta's structure and function (ie, nutrient transport and hormone and cytokine production) is a controversial subject. We hypothesized that maternal obesity is associated with morphologic, secretory, and nutrient-related changes and elevated levels of inflammation in the placenta. We collected samples of placental tissue from 2 well-defined groups of pregnant women from 2017 to 2019. We compared the 2 groups regarding placental cytokine and hormone secretion, immune cell content, morphology, and placental nutrient transporter expressions. Placenta were collected after caesarean section performed by experienced clinicians at Centre Hospitalier Intercommunal (CHI) of Poissy-Saint-Germain-en-Laye. The main inclusion criteria were an age between 27 and 37 years old, no complications of pregnancy, and a first-trimester body mass index of 18-25 kg/m2 for the nonobese (control) group and 30-40 kg/m2 for the obese group. In contrast to our starting hypothesis, we observed that maternal obesity was associated with (1) lower placental IL-6 expression and macrophage/leukocyte infiltration, (2) lower placental expression of GLUT1 and SNAT1-2, (3) a lower placental vessel density, and (4) lower levels of placental leptin and human chorionic gonadotropin production. These results suggest that the placenta is a plastic organ and could optimize fetal growth. A better understanding of placental adaptation is required because these changes may partly determine the fetal outcome in cases of maternal obesity.

SUN-513 Severe Hyperthyroidism in a Complete Molar Pregnancy

Hydatidiform mole (HM), a type of gestational trophoblastic disease (GTD), is a rare cause of clinical hyperthyroidism. The development of hyperthyroidism requires an elevation of HCG >100,000 mlU/mL for several weeks. Complete mole has a marked HCG elevation compared to partial mole thus presents with a higher incidence of thyrotoxicosis. Surgical uterine evacuation is the treatment of choice for HM. However, untreated hyperthyroidism can pose a risk for the development of thyroid storm and high-output cardiac failure in the perioperative period. To our knowledge, there are no specific guidelines for management at this time. We present a case of hyperthyroidism secondary to complete molar pregnancy successfully treated with propylthiouracil (PTU), potassium iodide (SSKI), and atenolol in the preoperative period.A 42-year-old female with history of migraines presented to her gynecologist with a 3-week history of lower abdominal cramping, vomiting, loss of appetite, and abnormal vaginal bleeding. She also endorsed a 6-pound weight loss, intermittent tachycardia, exertional dyspnea, and increased anxiety. Pregnancy test was positive, and ultrasound was concerning for GTD. Laboratory work up was significant for HCG 797,747 mIU/mL (< 5mlU/mL), TSH <0.005 mIU/mL (0.4-4.0 mlU/mL), Free T4 3.09 ng/dL (0.9-1.9 ng/dL), and Free T3 11.48 pg/dL (1.76-3.78 pg/dL). The patient was admitted to the hospital and started on PTU 100 mg Q6H, SSKI 200 mg TID following the first dose of PTU, and atenolol 25 mg daily. She underwent an uncomplicated D & C the next day. On post-op day 1, HCG decreased to 195,338 mIU/mL and Free T4 to 2.39 ng/dL. The patient was discharged on the aforementioned doses of PTU and atenolol. One-week follow-up labs showed HCG 8,917 mIU/mL and Free T4 1.22 ng/dL. Surgical pathology confirmed a complete hydatidiform mole. PTU was decreased to 50 mg TID. On post-op day 14, HCG had risen to 15,395 mIU/mL with onset of nausea and vomiting. Repeat Free T4 remained within reference range. Patient was taken back to surgery for a laparoscopic total hysterectomy with bilateral salpingectomy. Pathology confirmed an invasive hydatidiform mole. Two-week follow-up lab work showed HCG 155 mIU/mL, TSH 1.5 mIU/mL, and Free T4 1.19 ng/dL. PTU and atenolol were then discontinued.The development of hyperthyroidism in molar pregnancy is largely influenced by the level of HCG and usually resolves with treatment of GTD (1). However, it’s crucial to control thyrotoxicosis to avoid perioperative complications. This case also highlights the importance of monitoring HCG levels following a complete molar pregnancy due to an increased risk for invasive neoplasm.1. Walkington, L et al. “Hyperthyroidism and human chorionic gonadotrophin production in gestational trophoblastic disease.” British journal of cancer vol. 104,11 (2011): 1665-9. doi:10.1038/bjc.2011.139

A study on enhanced O-glycosylation strategy for improved production of recombinant human chorionic gonadotropin in Chinese hamster ovary cells

Human chorionic gonadotropin (hCG) is a glycoprotein hormone that exists as a heterodimer comprised of an α subunit and β subunit linked with disulfide bridges. The β subunit contains four O-glycosylation sites. Previous studies have found that the translation of mRNA to polypeptides of the β subunit was a severely limiting step for the expression of recombinant hCG protein in Chinese hamster ovary (CHO) cells. The effects of O-glycosylation on recombinant hCG protein expression were assessed by adding O-glycan precursors and overexpressing and knocking down key regulatory genes of O-glycan precursor synthesis and O-glycan sugar chain synthesis or hydrolases. The results indicated that O-glycosylation was indeed limiting in the expression of recombinant hCG protein, and N-acetylgalactosamine (GalNAc) was the major limiting precursor. Glutamine-fructose-6-phosphate transaminase 2 (Gfat2) and Uridine diphosphate-glucose pyrophosphorylase 2 (Ugp2), key regulatory genes of O-glycan precursor synthesis, were overexpressed. Ugp2 overexpression significantly increased the recombinant hCG protein level by 1.92 times compared to that of the control. The LC–MS/MS analysis and Phaseolus vulgaris leucoagglutinin (PHA-L) lectin blot analysis showed that Ugp2 overexpression significantly increased the total galactosylation levels of intracellular proteins and the O-glycosylation of recombinant hCG protein. The stability of the hCG protein to trypsin digestion was also enhanced. Ugp2 is the major limiting enzyme of the O-glycan precursor synthesis in recombinant hCG protein production. Furthermore, the effects and mechanisms of the key genes of O-glycan sugar chain synthesis and hydrolases such as polypeptide N-acetylgalactosaminyltransferase1 (Galnt1), Core 1 synthase, glycoprotein-N-acetylgalactosamine 3-beta-galactosyltransferase (C1galt1), O-linked N-acetylglucosamine transferase (Ogt) and Hexosaminidase (Hex), were evaluated. The results indicated that Galnt1 overexpression increased the recombinant hCG protein level by 1.57 times and improved the total galactosylation of intracellular proteins, O-glycosylation and the stability of recombinant hCG protein. Galnt1 is the major limiting enzyme of O-glycan sugar chain synthesis. Overexpression of Ugp2 and Galnt1 simultaneously improved the recombinant hCG protein level by 2.44 times, and both had synergistic effects. Based on the results of overexpression of Galnt1, the major limiting gene of O-Glycan chain synthesis, the precursors GalNAc and Gal were added and increased the recombinant hCG protein level by 3.68 times. This study revealed the major limiting factors of O-glycosylation of recombinant hCG protein in CHO cells and proposed an effective expression regulation strategy.

Maternal age and blastocyst quality do not influence the embryo production of hyperglycosylated human chorionic gonadotropin

Maternal age and blastocyst quality do not influence the embryo production of hyperglycosylated human chorionic gonadotropin

Solitary myofibroblastoma of the forearm presenting with ectopic [beta]-human Chorionic Gonadotrophin production.

Solitary myofibroblastoma of the forearm presenting with ectopic [beta]-human Chorionic Gonadotrophin production.

Quantitative proteomics suggest a potential link between early embryonic death and trisomy 16.

Activation of extracellular signal-regulated kinase (ERK) signalling, alteration of the uterine microenvironment and a reduction in human chorionic gonadotrophin production have been linked with fetal trisomy 16-induced early embryonic death (EED). However, the detailed biological mechanism of EED remains unclear. Using quantitative proteomics we successfully screened differentially expressed proteins in the villous tissues from patients with EED and fetal trisomy 16 (EEDT16), patients with EED but normal fetal chromosomes (EEDNC) and patients undergoing elective abortion with normal fetal chromosomes (EANC) as the reference group. Compared with the reference group, we identified 337 and 220 differentially expressed proteins in EEDT16 patients and EEDNC patients respectively; these were involved in critical biological processes including immune response, superoxide metabolism, inflammatory responses and so on. We found that differential expression of immunological function-related molecules, such as human leukocyte antigen-g (HLA-G), HLA-C, Fc Fragment Of IgG Receptor III (FcγR III), also named CD16, interleukin 18 (IL-18) and transforming growth factor β1 (TGF-β1), might induce EED in both EEDT16 and EEDNC patients. More severe immunological dysfunction was observed in EEDT16 patients than that in EEDNC patients. Furthermore, differential expression of implantation and invasion-related molecules, such as cytochrome b-245 light chain (CYBA), neutrophil cytosol factor 2 (NCF2), Mitogen-activated protein kinase kinase kinase 4 (MAP3K4), matrix metalloproteinase 2 (MMP2), MMP9 and tumour necrosis factor α (TNF-α) might induce EED in both EEDT16 and EEDNC patients, although more severe dysfunction in the implantation and invasion ability of villous tissues was observed in EEDT16 patients.

Versican silencing in BeWo cells and its implication in gestational trophoblastic diseases.

Versican is a proteoglycan known to interact with cells to influence their ability to proliferate, differentiate, migrate, invade and assemble extracellular matrix, with all of these cell functions present during placentation. In the placenta, cytotrophoblast cells have the ability to differentiate into the syncytiotrophoblast, a mechanism that is greatly increased in gestational trophoblastic diseases (GTD). Nevertheless, the molecular signaling underlying the increased syncytiotrophoblast differentiation are still being unveiled and may result in novel therapeutic targets for GTD. Versican expression was investigated to establish its differential expression among GTD (partial moles, complete moles, invasive moles and choriocarcinoma) and the possible functional outcomes from versican gene silencing. Tissue samples had their versican expression evaluated using immunohistochemistry and RT-PCR. BeWo cells were employed for versican silencing with siRNA and the efficiency was confirmed by RT-PCR, immunofluorescence and flow cytometry. Cell death and forskolin-induced syncytialization were analyzed by a morphological analysis and human chorionic gonadotropin (hCG) production using immunofluorescence. Versican V0 and V1 isoforms were mainly expressed in the syncytiotrophoblast and they were the most expressed in benign rather than in malignant tumors. BeWo cells also expressed V0 and V1 isoforms, but only in cells undergoing syncytial fusion. After versican silencing, cell death was greatly increased, whereas spontaneous and forskolin-induced syncytialization decreased as well as hCG production. Versican is differentially expressed in GTD and is important for hydatidiform moles pathophysiology, protecting trophoblast cells from death and playing a role in their differentiation and functionality.

Hyperthyroidism in a complete molar pregnancy with a mature cystic ovarian teratoma

BackgroundThe hallmark of gestational trophoblastic disease is the production of human chorionic gonadotropin (hCG) due to the hyperproliferation of extraembryonic trophoblast cells. Previous studies show hCG has thyrotropic action due to its structural similarity with thyroid stimulating hormone (TSH) molecules. Germ cell tumors represent 15–20% of all ovarian tumors and can be malignant or benign.Case presentationWe present a case of a 53-year old African American female with a history of hyperthyroidism secondary to a complete hydatidiform mole and an associated finding of a mature cystic ovarian teratoma. She presented with nausea, vomiting, nervousness, weight gain, abdominal pain and a b-hCG of greater than 450,000mIU/mL. A total abdominal hysterectomy with bilateral salpingo-oophorectomy was performed and curative for her symptoms. Lung nodules were noted with slight increases in b-hCG levels in the months following the surgery. Propranolol and methimazole were used to treat the acute hyperthyroid symptoms.ConclusionThis case presents the rare occurrence of a complete hydatidiform mole causing hyperthyroidism and an associated finding of a mature cystic teratoma. It also highlights the importance of monitoring b-hCG levels following a complete molar pregnancy due to an increased risk of choriocarcinoma.

Impaired serum inhibin-B and number of germ cells in boys with cryptorchidism following heavily gestational maternal smoking

PurposeA meta-analysis including 11,900 cases showed that maternal gestational smoking was associated with increased risk of cryptorchidism. The aim of study was to investigate whether a hormone profile of cryptorchid boys and a supplementing histopathological evaluation of testicular biopsies could add detailed knowledge to the impact of maternal gestational smoking on pathogenesis of cryptorchidism. Methods601 cryptorchid boys aged 4 months to 14 years old were included. Because normal hormones have a pronounced age dependency, we compared results from boys whose mothers had smoked heavily (>10 cigarettes/day) during pregnancy with age matched cryptorchid controls of nonsmoking mothers (1:6). We studied: birthweight, germ-cell number/tubular cross section, frequency of germ cells positive for placental-like alkaline phosphatase (PLAP), gonadotropins and inhibin-B. Results501 boys were sons of nonsmokers, 72 boys of intermittent smokers and 28 boys of heavy smokers. 39%, 44% and 61% respectively had bilateral cryptorchidism. Compared to age-matched cryptorchid controls of nonsmoking mothers, sons of heavy smokers had lower birthweight (p = 0.006), germ-cell number/tubular cross section (p = 0.009), frequency of germ cells positive for PLAP (p = 0.037) and inhibin-B (p = 0.042). ConclusionsAll findings could be associated with placental dysfunction with altered human chorionic gonadotropin production well described in women smoking during pregnancy. Type of studyPrognosis study (prospective cohort study with >80% follow-up). Level of evidenceLevel 1.

Regulation of human trophoblast cell syncytialization by transcription factors STAT5B and NR4A3.

In human trophoblast cells, cyclic AMP or its inducer forskolin (FSK) activates two downstream signaling molecules, protein kinase A (PKA) and exchange protein directly activated by cAMP (EPAC), both of which induce syncytialization, cell fusion, and the production of human chorionic gonadotropin (hCG) and progesterone. However, a transcription factor other than GCM1 and molecular mechanisms associated with these events have not been well characterized. To identify novel transcription factors involved in syncytialization of cAMP-stimulated human choriocarcinoma BeWo cells, the microarray analysis was performed with RNAs extracted from PKA- or EPAC-selective cAMP analog-stimulated BeWo cells, from which two up-regulated transcription factors, STAT5 and NR4A3, were found. The knockdown of STAT5B decreased FSK-induced cell fusion and the expression of syncytialization markers, CGB, syncytin1, syncytin2, GCM1, and OVOL1, but NR4A3 knockdown increased FSK-induced cell fusion and the expression of CGB and syncytin2. These findings indicated that cAMP-PKA up-regulated STAT5B, followed by increase in syncytin2 expression through GCM1 and OVOL1, resulting in cell fusion and hCG production, while cAMP-PKA-up-regulated NR4A3 could decrease syncytin2 expression, and suggested that both positive and negative effects of STAT5B and NR4A3, respectively, are required to control the degree of syncytialization in human trophoblast cells.

Daidzein induces choriocarcinoma cell apoptosis in a dose-dependent manner via the mitochondrial apoptotic pathway.

Choriocarcinoma is a malignant gestational trophoblastic disease and relapse or drug resistance occurs in ~25% of gestational trophoblastic tumors. Cell apoptosis serves a role in the progression from hydatidiform mole to persistent gestational trophoblastic disease. It has been demonstrated that daidzein [7‑hydroxy‑3‑(4‑hydroxyphenyl)‑4H‑chromen‑4‑one] may induce apoptosis in a number of cancer types via the mitochondrial apoptotic pathway by altering the B‑cell lymphoma (Bcl)‑2/Bcl‑2 associated X, apoptosis regulator (Bax) ratio, and activating the caspase cascade. Daidzein also serves a role in regulation of production of human chorionic gonadotropin in trophoblast cells and inhibition of cell proliferation. However, few reports have been published regarding the effect of daidzein on apoptosis in choriocarcinoma. Therefore, in the present study, JAR and JEG‑3 human gestational choriocarcinoma cells were used to investigate the effect of daidzein on apoptosis of choriocarcinoma cells. Treatment with daidzein for 48h reduced cell viability in a dose‑dependent manner. The percentages of early and late apoptotic cells also increased following treatment with daidzein in a dose‑dependent manner, with the number of late apoptotic cells increasing more prominently. Furthermore, treatment with daidzein led to apoptosis‑associated alterations in nuclear morphology of JAR and JEG-3 cells. Expression levels of cleaved poly(ADP‑ribose) polymerase, cleaved caspase‑3 and cleaved caspase‑9 increased following treatment with daidzein, whereas the Bcl‑2/Bax ratio decreased in a dose‑dependent manner. In conclusion, the results of the present study demonstrate that daidzein may induce apoptosis of choriocarcinoma cells in a dose‑dependent manner via the mitochondrial apoptotic pathway.

The usefulness of testosterone administration in identifying false-positive elevation of serum human chorionic gonadotropin in patients with germ cell tumor.

The pituitary production of human chorionic gonadotropin (hCG) can cause false-positive results during or after germ cell tumor (GCT) treatment. Because hypogonadism leads to pituitary hCG production, testosterone administration test (TAT) has been recommended for pituitary hCG diagnosis. However, little is known about its efficacy for the discrimination of pituitary hCG as detected by currently used hCG assays in treatment of GCT. We conducted a retrospective multicenter study to determine the usefulness of TAT. The study included 60 patients who underwent TAT for the discrimination of pituitary hCG. In principle, serum hCG levels were measured 1week after testosterone enanthate administration (250mg). When the serum hCG levels decreased below the normal upper range, the results of TAT were determined positive. In this case, the elevated hCG was considered to be derived from pituitary and not from GCT. Serum hCG levels were normalized after TAT in 36 of 60 patients (60%). Before TAT, the hCG levels were below 1.0IU/L in 13 patients (36%), 1.0-1.9IU/L in 11 (31%), 2.0-2.9IU/L in 7 (19%), and >3.0IU/L in 5 (14%) of TAT-positive patients. Of them, 28 (78%) patients were successfully managed without further treatment with chemotherapy after TAT. Pituitary hCG was associated with higher levels of LH and not necessarily associated with low levels of testosterone. Determining the TAT status of patients was effective in discriminating pituitary hCG production.

Human chorionic gonadotropin (HCG) as a biomarker in sarcoma: A systematic review.

e22503 Background: Human chorionic gonadotropin (HCG) has been reported in soft tissue sarcoma and osteosarcoma. In some patients, the ectopic production of HCG can lead to delays in diagnosis due to concern for pregnancy or germ cell tumors. Methods: We performed a systematic review of the scientific literature in English using PubMed to categorize cases of sarcoma search terms were HCG, sarcoma, human chorionic gonadotropin and osteosarcoma. Cases of sarcoma associated with HCG production were identified and further categorized by type and grade of tumor. Initial HCG, Peak HCG, response to therapy and case outcome were collected when available. We also included a case seen at our institution of osteosarcoma with ectopic HCG production. Results: We identified 21 manuscripts and 23 cases of patients with both sarcoma and elevated serum HCG. 10/23 patients were male (43%).The most frequent type of sarcoma was osteosarcoma with 9/23 (39%) of cases, leiomyosarcoma was identified in 7/23 (30%) and liposarcoma in 3/23 (13%) of cases. When described, all tumors were high grade or poorly differentiated. 20/23 tumors (87%) had HCG expression of the tumor by immunohistochemistry. Mean initial HCG was 3607 with a peak value of 6188. In 19 cases, HCG was used to monitor disease after treatment and 14/19 (74%) responded with a decrease in HCG after treatment. Of responders, 3 patients were considered cured and 2 were put into remission, while none of the patients without response survived. Overall, outcomes were poor with 16/23 (70%) deaths within one year. Conclusions: Sarcomas that secrete HCG tend to be high grade, poorly differentiated and appear to portend a poor prognosis. When expressed, HCG can be used as a marker of disease and response to therapy. Oncologists should consider checking HCG as a potentially low-cost marker of prognosis and response to treatment in patients with sarcoma. [Table: see text]

Activation of Peroxisome Proliferator-Activated Receptor Gamma and Disruption of Progesterone Synthesis of 2-Ethylhexyl Diphenyl Phosphate in Human Placental Choriocarcinoma Cells: Comparison with Triphenyl Phosphate.

2-Ethylhexyl diphenyl phosphate (EHDPP), an organophosphate flame retardant (OPFR), is frequently detected in human blood. In this study, the sensitive dual-luciferase reporter gene assay and molecular docking were used to investigate the activation of EHDPP to human peroxisome proliferator-activated receptor gamma (PPARG). Results show that EHDPP exhibited stronger PPARG activation (EC20: 2.04 μM) than triphenyl phosphate (TPhP) (EC20: 2.78 μM). EHDPP upregulated the gene expression of 3β-hydroxysteroid dehydrogenase type 1 (3β-HSD1) in human placental choriocarcinoma cells in a dose-dependent manner, and the lowest observable effective concentration was 10 μM, lower than that of TPhP (20 μM). EHDPP significantly altered progesterone secretion at a lower concentration (10 μM) than that of TPhP (20 μM), and both EHDPP and TPhP significantly promoted human chorionic gonadotropin (hCG) production at 20 μM. Furthermore, inactivation of PPARG by either a pharmacological inhibitor (GW9662) or small interfering RNA (siRNA) abolished the change in progesterone secretion and gene expression in the cells exposed to EHDPP, suggesting that the PPARG signaling pathway plays a role in the upregulation of progesterone by the two OPFRs. This is the first report to show that OPFRs can alter the biosynthesis of progesterone in the placenta, which could affect female reproduction and fetal development.

Circulating Cell Free DNA in the Diagnosis of Trophoblastic Tumors

Gestational trophoblastic neoplasia (GTN) represents a group of diseases characterized by production of human chorionic gonadotropin (hCG). Since non-gestational tumors may occasionally secrete hCG, histopathological diagnosis is important for appropriate clinical management. However, a histopathological diagnosis is not always available. We therefore investigated the feasibility of extracting cell free DNA (cfDNA) from the plasma of women with GTN for use as a “liquid biopsy” in patients without histopathological diagnosis. cfDNA was prepared from the plasma of 20 women with a diagnosis of GTN and five with hCG-secreting tumors of unknown origin. Genotyping of cfDNA from the patient, genomic DNA from her and her partner and DNA from the tumor tissue identified circulating tumor DNA (ctDNA) (from 9% to 53% of total cfDNA) in 12 of 20 patients with GTN. In one case without a tissue diagnosis, ctDNA enabled a diagnosis of GTN originating in a non-molar conception and in another a diagnosis of non-gestational tumor, based on the high degree of allelic instability and loss of heterozygosity in the ctDNA. In summary ctDNA can be detected in the plasma of women with GTN and can facilitate the diagnosis of both gestational and non-gestational trophoblastic tumors in cases without histopathological diagnosis.