SFRP2 suppresses trophoblast cell migration by inhibiting the Wnt/ß­catenin pathway.

The present study investigates the role of Secreted Frizzled­Related Protein 2 (SFRP2) in trophoblast cells, a key factor in preeclampsia (PE) progression. Elevated levels of Secreted Frizzled­Related Protein 1/3/4/5 (SFRP1/3/4/5) are associated with PE, but the role of SFRP2 is unclear. We analyzed SFRP2 expression in PE placental tissue using the GSE10588 dataset and overexpressed SFRP2 in JEG­3 cells via lentiviral transfection. The viability, migration, apoptosis, and proliferation of SFRP2­overexpressing JEG­3 cells were assessed using Cell Counting Kit­8, Transwell assays, flow cytometry, and EdU staining. Additionally, we evaluated the impact of SFRP2 overexpression on key proteins in the Wnt/ß­catenin pathway and apoptosis markers (Bax, cleaved­caspase 3, BCL­2, MMP9, E­cadherin, Wnt3a, Axin2, CyclinD1, c­Myc, p­ß­catenin, ß­catenin, phosphorylated Glycogen Synthase Kinase 3 beta (p­GSK3ß), and GSK3ß) through western blotting. Results showed high SFRP2 mRNA and protein expression in PE placenta and JEG­3 cells post­transfection. SFRP2 overexpression significantly reduced JEG­3 cell viability, proliferation, and migration, while increasing apoptosis. It also altered expression levels of Wnt pathway proteins, suggesting SFRP2's potential as a therapeutic target for PE by inhibiting trophoblast cell migration through the Wnt/ß­catenin signaling cascade.

METTL3-mediated lncRNA HOXD-AS1 stability regulates inflammation, and the migration and invasion of trophoblast cells via the miR-135a/ ß-TRCP axis.

Background: Preeclampsia (PE) is a serious pregnancy-specific syndrome associated with the inadequate invasion of trophoblast cells and inflammation of the uterus. A previous study found that lncRNA HOXD-AS1 promotes PE. However, its regulatory network requires additional exploration. Methods: HOXD-AS1-targeted miRNAs and genes were predicted by different databases in a bioinformatics analysis. The expression HOXD-AS1 and its potential m6A methylase (METTL3) were detected in placentas from healthy female patients with PE. The targeting relationship and role of the HOXD-AS1/miR-135a/ß-TRCP axis in trophoblast cells were demonstrated by overexpression/knockdown assays. The levels of ß-TRCP downstream pathway proteins IκBα, NF-κB, and p65 were measured. The levels of inflammatory factors in cell supernatants were detected by ELISA. To verify the molecular mechanism of ß-TRCP in PE, IκBα was co-overexpressed in ß-TRCP in trophoblast cells. Results: The levels of METTL3, HOXD-AS1, and ß-TRCP were elevated in PE placental tissues, while miR-135a levels were reduced. MiR-135a was found to be targeted by HOXD-AS1, and HOXD-AS1 expression was maintained at a high level by METTL3-mediated m6A methylation. Overexpression of METTL3, HOXD-AS1, and ß-TRCP, and knockdown of miR-135a in HTR-8/SVneo cells all inhibited cell invasion and migration, and promoted apoptosis and the secretion of inflammatory factors. Knockdown of METTL3, HOXD-AS1, and ß-TRCP, and overexpression of miR-135a had the opposite effects. Furthermore, IκBα expression was negatively associated with ß-TRCP expression, and the levels of NF-κB, p65, and NLRP3 were positively regulated by ß-TRCP. A high level of ß-TRCP expression attenuated the effect of HOXD-AS1 knockdown in trophoblast cells. Conclusion: METTL3 functioned to maintain a high level of HOXD-AS1 expression in PE, which influenced inflammation and the migration and invasion of trophoblast cells via the miR-135a/ß-TRCP axis and NF-κB pathway.

Effect of modified levonorgestrel-releasing intrauterine system in human adenomyosis with heavy menstrual bleeding.

AIM: The levonorgestrel-releasing intrauterine (LNG-IUS) system is an effective primary treatment for adenomyosis; however, it has high expulsion rates. We aimed to modify the system-allowing affixion to the myometrium-and evaluate the expulsion rate, effectiveness, and side effects in patients with adenomyosis and heavy menstrual bleeding. METHODS: This study included patients with adenomyosis and heavy menstrual bleeding who underwent implantation of: a modified LNG-IUS (experimental group, n = 47); and the original system after gonadotropin-releasing hormone agonist treatment (control group, n = 47), between January 2014 and April 2016. RESULTS: In the experimental group, two device expulsions occurred 12-18 months postimplantation. In the remaining 45 patients, the system was safely removed after the 60-month validity period, and no extrauterine device movement or infection occurred. In the control group, downward displacement and expulsion of the device occurred in eight (17%) patients within 60 months. The 5-year total expulsion rates were 4.3% and 17.0% in the experimental and control groups, respectively (p = 0.045). There were significant changes in the pretreatment severity of dysmenorrhea, menstrual volume, uterine volume (cm3 ), and hemoglobin level in each group compared with after 1 year (p < 0.01 in all groups). The severity of dysmenorrhea, menstrual volume, uterine volume, and hemoglobin level after 1 year were similar between the two groups (p > 0.05 in all groups). CONCLUSIONS: Use of the modified LNG-IUS is a safe, cost-effective, and simple method for reducing the downward movement and expulsion rate in patients with adenomyosis and heavy menstrual bleeding.

Fas regulates the apoptosis and migration of trophoblast cells by targeting NF-κB.

Placental trophoblast apoptosis is a major pathological feature of preeclampsia. Fas has been reported to be highly expressed in the placentas of patients with preeclampsia. However, the role and underlying mechanisms of Fas in the pathogenesis of preeclampsia have not been elucidated. In the present study, the expression of Fas in JAR human choriocarcinoma cells was overexpressed and knocked down to determine the function and possible mechanism of Fas in trophoblast cells in the progression of preeclampsia. The results of flow cytometry, Cell Counting Kit-8 and Transwell assays indicated that the overexpression of Fas promoted apoptosis, suppressed viability and impaired the migration of the human trophoblast cells. In addition, western blotting revealed that the overexpression of Fas increased the expression of nuclear factor kB (NF-kB), Bax, tumor necrosis factor α (TNF-α) and interleukin-2 (IL-2), and decreased the expression of Bcl-2 at the protein level in trophoblast cells. By contrast, the knockdown of Fas decreased the apoptosis of trophoblast cells and increased their viability and migration. In addition, the knockdown of Fas suppressed the expression of NF-κB, Bax, TNF-α and IL-2, and increased the expression of Bcl-2. Notably, the overexpression of NF-κB p65 attenuated the Fas knockdown-induced inhibition of apoptosis and acceleration of migration of the trophoblast cells. The overexpression of NF-κB in trophoblast cells also reversed the reduction in Bax expression and increase in Bcl-2 expression induced by Fas knockdown in trophoblast cells. These results indicate that Fas regulates the apoptosis and migration of trophoblast cells by targeting NF-κB, which suggests that the silencing of Fas is a promising therapeutic strategy for preeclampsia.

Significance of highly phosphorylated insulin-like growth factor binding protein-1 and cervical length for prediction of preterm delivery in twin pregnancies.

BACKGROUND: A twin pregnancy can carry greater risks than singleton pregnancies. About 60 in 100 twin pregnancies result in spontaneous birth before 37 wk, which is associated with several complications in the premature babies. Clinical detection of biomarkers may help to predict the possibility of premature birth so that corresponding interventions can be given to the pregnant women in a timely manner, in order to reduce the risk of preterm birth and improve the outcomes of the newborn infants. AIM: To explore the clinical value of transvaginal ultrasound measurement of cervical length combined with insulin-like growth factor binding protein-1 (IGFBP-1) hyperphosphorylation in cervical secretions as predictors of preterm delivery in twin pregnancies. METHODS: A total of 254 pregnant women with twin pregnancies, who were admitted to Hainan General Hospital and underwent maternity examination, were selected as the study subjects from January 2015 to December 2018. All participants received transvaginal ultrasound measurement of cervical length and phosphorylated IGFBP-1 (phIGFBP-1) test between 24 and 34 wk gestation. The pregnancy outcomes were analyzed. RESULTS: Of the women with a positive phIGFBP-1 test result, preterm birth rate was higher in those with a cervical length ≤ 25 mm than those with a cervical length > 25 mm (all P < 0.05). Similarly, in women with a negative phIGFBP-1 test result, preterm birth rate was higher in those with a cervical length ≤ 25 mm than those with a cervical length > 25 mm (all P < 0.05). The sensitivity, specificity, and positive and negative predictive values of the phIGFBP-1 test combined with the cervical length test were 95.71%, 91.21%, 95.12% and 92.22%, respectively, for the prediction of preterm birth. CONCLUSION: Cervical length combined with phIGFBP-1 tests is of value for the prediction of outcomes of preterm delivery in twin pregnancies.