Trophoblast Cell Proliferation Research Articles

A Novel Circular RNA CircBRAP May Be Used as an Early Predictor of Preeclampsia and Its Potential Mechanism.

Preeclampsia (PE), a pregnancy-related multisystem syndrome, is one of the leading causes of maternal and fetal mortality worldwide. The aim of this study was to combine the plasma protein soluble Fms-related tyrosine kinase 1 (sFLT1) levels with uterine artery Doppler ultrasound findings and CircBRAP levels during the first trimester to predict the occurrence of preeclampsia and to explore the potential mechanism by which CircBRAP functions in preeclampsia. Here, we used qRT-PCR to investigate the expression of CircBRAP in forty-nine pairs of plasma specimens and placental tissues from preeclampsia patients and control subjects. The uterine artery (UtA) pulsatility index (PI) was measured using four-dimensional color Doppler ultrasound, and the sFLT1 levels were evaluated by human immunoassay. Exogenous upregulation or downregulation of CircBRAP expression in TEV-1 trophoblast cells was performed to investigate the role of CircBRAP in cell biological behavior. Mechanistically, luciferase reporter, RNA immunoprecipitation (RIP), and biotin-coupled RNA pull-down assays were conducted to verify the relationship between CircBRAP and sFLT1 in TEV-1 cells. The results showed that the predictive power was strengthened when the plasma sFLT1 and CircBRAP levels were combined with the UtA-PI to predict preeclampsia occurrence. Our study also revealed that CircBRAP may regulate miR-106b and the HIF-2α axis to modulate the proliferation, invasion, and apoptosis of TEV-1 trophoblast cells. In summary, placenta-derived CircBRAP in plasma may be a novel biomarker for preeclampsia that, together with plasma sFLT1 levels and uterine artery Doppler ultrasound findings, can more effectively predict preeclampsia, and CircBRAP may play a potential role in preeclampsia.

CircPAPPA Regulates the Proliferation, Migration, Invasion, Apoptosis, and Cell Cycle of Trophoblast Cells Through the miR-3127-5p/HOXA7 Axis.

Abnormal function of trophoblast cells is one of the important causes of preeclampsia (PE). Circular RNA (circRNA) is thought to be involved in the regulation of various diseases progression, including PE. However, the role of circRNA pregnancy-associated plasma protein A (circPAPPA) in PE is less studied. The expression levels of circPAPPA, miR-3127-5p, and homeobox A7 (HOXA7) were determined by quantitative real-time PCR. Cell proliferation was evaluated using MTT assay and colony formation assay. Besides, flow cytometry was used to detect cell apoptosis and cell cycle distribution. In addition, the interaction between miR-3127-5p and circPAPPA or HOXA7 was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation assay. CircPAPPA was lowly expressed in the placental tissues of PE patients. Knockdown of circPAPPA inhibited proliferation, migration, and invasion, while induced apoptosis and cell cycle arrest in trophoblast cells. MiR-3127-5p could be targeted by circPAPPA, and its inhibitor reversed the effect of circPAPPA silencing on the biological function of trophoblast cells. Moreover, HOXA7 was a target of miR-3127-5p. HOXA7 overexpression reversed the effect of miR-3127-5p on the biological function of trophoblast cells. Our research indicated that circPAPPA positively regulated the biological function of trophoblast cells to mediate the progression of PE by miR-3127-5p/HOXA7 axis, which suggested that circPAPPA might be a potential biomarker for PE.

Interleukin-17 promotes proliferation, migration, and invasion of trophoblasts via regulating PPAR-γ/RXR-α/Wnt signaling

ABSTRACT To investigate the effect of Interleukin 17 (IL-17) on the invasive capacity of trophoblast cells and the underlying mechanism, we collected placental tissues samples from pregnant women with preeclampsia (PE) and healthy pregnant women. The expression levels of IL-17 mRNA and protein in tissue samples were determined using qRT-PCR and Western blot, respectively. Cell viability and cell proliferation was determined using CCK-8 assay, and colony formation assay, respectively. Cell migration and invasion capacity were determined using transwell cell migration assay. Our results showed that the mRNA expression of IL-17 was significantly increased in PE patients and may be used as a sensitive biomarker for PE (P < 0.01). IL-17 overexpression promoted cell viability, migration, and invasion of human extravillous trophoblast cell line, HTR8/SVneo; however, IL-17 knockdown inhibited these effects. Additionally, IL-17 activated PPAR-γ/RXR-α signaling pathway, which promoted proliferation, migration, and invasion of trophoblast cells. Moreover, PPAR-γ/RXR-α heterodimers activated Wnt signaling. In conclusion, our study provides evidence that IL-17 is overexpressed in PE and promotes proliferation, migration and invasion of trophoblast cells via activating PPAR-γ/RXR-α/Wnt signaling.

Overexpressed IncRNA GATA3-AS1 in Preeclampsia and Its Effects on Trophoblast Proliferation and Migration by the miR-488-3p/ROCK1 Axis.

Recently, dysregulation of long noncoding RNAs (lncRNAs) has been reported to be involved in the pathogenesis of preeclampsia (PE). Here, the role and molecular mechanisms of lncRNA GATA3 antisense RNA 1, GATA3-AS1 in PE were explored. The expression of GATA3-AS1, miR-488-3p and Rho-associated coiled-coil-containing protein kinase 1 (ROCK1) in placental tissues from patients with PE was measured by reverse transcription quantitative PCR (RT-qPCR). Proliferation, migration, invasion, and apoptosis of trophoblast cells were examined by 5-ethynyl-2'-deoxyuridine (EdU), wound healing, Transwell, and flow cytometry analyses. The subcellular localization of GATA3-AS1 in trophoblast cells was determined by fluorescent hybridization (FISH) assay. The interactions among GATA3-AS1, miR-488-3p and ROCK1 were identified by luciferase reporter and RNA pulldown assays. Our results showed that GATA3-AS1 and ROCK1 were overexpressed while miR-488-3p was downregulated in placental samples with PE. Functionally, GATA3-AS1 overexpression promoted trophoblast cell apoptosis and inhibited cell proliferation, migration, and invasion. Mechanically, GATA3-AS1 acted as a molecular sponge of miR-488-3p and miR-488-3p targeted ROCK1 in trophoblast cells. In rescue assays, ROCK1 overexpression or miR-488-3p downregulation reversed the effects of GATA3-AS1 silencing on trophoblast cell phenotypes. GATA3-AS1 is overexpressed in PE and promotes PE progression by the miR-488-3p/ROCK1 axis.

MiR-515-5p Suppresses Trophoblast Cell Invasion and Proliferation Through XIAP Regulation in Preeclampsia

miR-515-5p Suppresses Trophoblast Cell Invasion and Proliferation Through XIAP Regulation in Preeclampsia

Circ_0007445 Inhibits Trophoblast Cell Proliferation, Migration and Invasion by Targeting HTRA1 Through the Regulation of miR-4432 in Pre-Eclampsia

Circ_0007445 Inhibits Trophoblast Cell Proliferation, Migration and Invasion by Targeting HTRA1 Through the Regulation of miR-4432 in Pre-Eclampsia

Re-expression of circ_0043610 contributes to trophoblast dysfunction through the miR-558/RYBP pathway in preeclampsia.

An increasing number of data have shown the pathogenesis of preeclampsia (PE) involves circular RNA (circRNA). The study aims to investigate the function and the potential mechanism of circ_0043610 in PE. The study was performed on two human placental trophoblastic cell lines (JEG-3 and HTR-8/SVneo). The expression of circ_0043610, microRNA-558 (miR-558), and RING1 and YY1 binding protein (RYBP) was detected by quantitative real-time polymerase chain reaction. The protein levels of N-cadherin, E-cadherin, and RYBP were assessed by Western blotting. Cell viability, proliferation, apoptosis, invasion, and migration were evaluated by cell counting kit-8, 5-Ethynyl-29-deoxyuridine, flow cytometry analysis, transwell invasion assay, and wound-healing assay, respectively. Dual-luciferase reporter assay, RNA immunoprecipitation assay, and RNA pull-down assay were performed to identify the associations among circ_0043610, miR-558, and RYBP. Compared with normal placental controls, the increased expression of circ_0043610 and RYBP and the decreased miR-558 expression were detected in PE placental tissues. The overexpression of circ_0043610 led to decreased trophoblast cell proliferation, invasion, and migration but increased cell apoptosis. Mechanistically, circ_0043610 acted as a miR-558 sponge, and miR-558 bound to RYBP. Besides, miR-558 introduction remitted circ_0043610-mediated effects in JEG-3 and HTR-8/SVneo cells. Moreover, RYBP participated in the regulation of miR-558 on trophoblast cell behaviors. Further, the ectopic expression of circ_0043610 led to RYBP upregulation through miR-558. Circ_0043610 induced RYBP production to promote trophoblast dysfunction by binding to miR-558 in PE.

Protein tyrosine phosphatase receptor type O (PTPRO) knockdown enhances the proliferative, invasive and angiogenic activities of trophoblast cells by suppressing ER resident protein 44 (ERp44) expression in preeclampsia

ABSTRACT Preeclampsia (PE), a pregnancy-specific syndrome, is the primary cause of maternal mortality. This work was designed to investigate the specific functions of PTPRO/ ERp44 in the biological behaviors of trophoblast cells and elucidate the underlying molecular mechanism. Constructed siRNA-PTPRO and ERp44 overexpression plasmids were transfected into HTR-8/SVneo and JEG-3 cells for further functional experiments. Subsequently, the proliferation and invasion of trophoblast cells were identified by performing CCK-8, flow cytometry and transwell assay. In addition, tube formation assay was employed to estimate the angiogenic ability of HUVECs incubated with the conditioned media (CM) of HTR-8/SVneo or JEG-3 cells. Importantly, the interaction between PTPRO and ERp44 was analyzed through Co-IP. In the current investigation, it was discovered that downregulation of PTPRO notably facilitated the proliferation and invasion of trophoblast cells and induced a stronger in vitro angiogenesis. Moreover, PTPRO interacted with ERp44 to regulate ERp44 expression. ERp44 overexpression suppressed the proliferative, invasive and angiogenic activities of trophoblast cells. As a result, functions of PTPRO knockdown in the biological behaviors of trophoblast cells were partially abrogated upon elevation of ERp44. To sum up, this current research systematically evidenced that PTPRO could regulate the biological behaviors of trophoblast cells by modulating ERp44. Findings may contribute to a novel therapeutic strategy for PE.

Circ_0037078 promotes trophoblast cell proliferation, migration, invasion and angiogenesis by miR-576-5p/IL1RAP axis.

Preeclampsia (PE) is a common hypertensive disorder of pregnancy. Recent studies have suggested that circular RNAs (circRNAs) play a pathological role in PE. Herein, this study aimed to investigate the action and mechanism of circ_0037078 in PE process. The quantitative real-time PCR (qRT-PCR) and Western blot were used to determine the expression levels of RNAs and genes. Cell proliferation, migration, invasion and angiogenesis were evaluated by using cell counting kit-8 (CCK-8), colony formation, transwell, and tube formation assays, respectively. The target relation between miR-576-5p and IL1RAP (Interleukin-1 receptor accessory protein) and circ_0037078 was predicted by bioinformatics analysis and verified by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Circ_0037078 expression was higher in placental tissues of patients with PE than that of normal control. Knockdown of circ_0037078 led to an enhancement of the proliferation, migration, invasion, and angiogenesis in trophoblast cells. Mechanistically, circ_0037078 acted as a sponge for miR-576-5p, thus elevating the expression of IL1RAP, which was targeted by IL1RAP. Further rescue experiments suggested that miR-576-5p inhibition reversed the effects of circ_0037078 knockdown on above behaviors of trophoblast cells. Moreover, miR-576-5p overexpression enhanced the proliferative, migratory, invasive, angiogenic phenotypes of trophoblast cells, which were attenuated by IL1RAP up-regulation. Circ_0037078 knockdown promotes trophoblast cell proliferation, migration, invasion, and angiogenesis in vitro by miR-576-5p/IL1RAP axis, providing a novel insight into the etiology of PE.

BPDE induces human trophoblast cell ferroptosis by up-regulating iron metabolism and promoting GPX4 proteasomal degradation

Human trophoblast cells play important role in embryo-fetal development. However, trophoblast cells are sensitive to environmental carcinogens. Recently, we have discovered that BPDE inhibits trophoblast cell migration, invasion and proliferation, and also increases trophoblast cell apoptosis. Ferroptosis is a newly identified iron-dependent non-apoptotic programmed cell death. Whether BPDE might induce trophoblast cell ferroptosis is completely unknown. In this work, we have discovered that ferroptosis does occur in BPDE-treated human trophoblast cells. Iron metabolism up-regulates intracellular free Fe2+ level, produces excessive ROS (reactive oxygen species), and accelerates lipid peroxidation. GPX4 efficiently eliminates ROS and inhibits lipid peroxidation. Thus, ferroptosis is well suppressed due to the balance of these two pathways in healthy trophoblast cells. However, BPDE exposure could promote iron metabolism, increase intracellular free Fe2+ level, produce excessive ROS, and result in lipid peroxidation. Meanwhile, BPDE also down-regulates GPX4 expression level by promoting its proteasomal degradation, which eventually produces excessive ROS and induces lipid peroxidation. Thus, BPDE exposure induces trophoblast cell ferroptosis by unbalancing these two pathways, providing new insight in understanding BPDE-induced dysfunctions of human trophoblast cells.

LncRNA DANCR promotes the migration an invasion and of trophoblast cells through microRNA-214-5p in preeclampsia

ABSTRACT Studies have shown that lncRNA DANCR is down-regulated in placental tissues of patients with preeclampsia (PE). The aim of this study was to explore the effect of lncRNA DANCR on trophoblast cells as well as its acting mechanism. We disrupted or overexpressed lncRNA DANCR in trophoblast cells HTR-8/SVneo and JEG-3 and detected the associated cellular functional changes by MTT, flow cytometry, Transwell experiment, and scratch experiment. The results showed that overexpression of lncRNA DANCR significantly increased the proliferation, invasion, migration, and EMT process of trophoblast cells. Interfering with lncRNA DANCR showed the opposite result. Further, the targeted interaction between lncRNA DANCR and miR-214-5p was confirmed by the dual-luciferase reporter gene assay. In addition, the expression of PI3K/AKT signaling pathway-related proteins was analyzed by Western blot. Overexpression of lncRNA DANCR can increase the phosphorylation of PI3K/AKT protein and activate this signaling pathway. In conclusion, the enforcing of lncRNA DANCR activates the activation of the PI3K/AKT pathway by down-regulating miR-214-5p, and promotes the migration and invasion of chorionic trophoblast cells. This provides a potential new target for PE therapy.

The MEG3 lncRNA promotes trophoblastic cell growth and invasiveness in preeclampsia by acting as a sponge for miR-21, which regulates BMPR2 levels.

Preeclampsia (PE) is one of the leading causes of maternal morbidity and mortality in pregnant women. This study aimed to investigate the potential impact and regulatory mechanisms of bone morphogenetic protein receptor 2 (BMPR2) on the progression of PE. We obtained placental tissues from pregnant women with PE and normal pregnant women, and the results showed that BMPR2 was expressed at low levels in the tissue from PE women. Genetic knockdown of BMPR2 increased the proliferation and invasion of cultured trophoblast cells, whereas its overexpression reduced these characteristics. Bioinformatics analysis and luciferase reporter gene assays confirmed that BMPR2 is a direct target of miR-21. Overexpression of a miR-21 inhibitor promoted the growth and invasiveness of trophoblast cells, whereas the opposite results were observed for the miR-21 mimic. Furthermore, miR-21 was sponged by the lncRNA MEG3, and shRNA inhibition of MEG3 reduced trophoblast cell growth and invasiveness. miR-21 was upregulated in the tissues from PE women, whereas MEG3 was downregulated, and the two were negatively correlated. Collectively, this study demonstrates that the lncRNA MEG3 acts as a sponge for miR-21, which regulates BMPR2 expression and promotes trophoblast cell proliferation and invasiveness, thereby preventing the development of PE. These findings provide novel insight into a targeted therapy that could be used to treat or prevent the development of PE.

The potential risk factors of placenta increta and the role of octamethylcyclotetrasiloxane.

The study aimed to investigate the potential risk factors for the placenta accreta spectrum (PAS), determine the predictive value of a diagnostic model, and evaluate the effects of octamethylcyclotetrasiloxane (OMCTS) on trophoblast proliferation and migration. This case-control study included 244 pregnant women with PAS and 327 normal pregnant women who visited Guangzhou Women and Children's Medical Centre, China, from January 2014 to December 2017. Blood was collected from 42 women with PAS and 77 controls, and plasma specimens were analyzed by gas chromatography-time-of-flight mass spectrometry. In addition, the proliferation and migration of trophoblast cells were examined after treatment with OMCTS. We found an association between the risk of PAS and clinical factors related to fasting blood glucose levels (BS0, OR = 5.78), as well as factors related to endometrial injury [history of cesarean section (OR = 179.59), uterine scarring (OR = 68.37), and history of abortion (OR = 5.66)]. Equally important, pregnant women with PAS had significantly higher plasma OMCTS concentrations than controls. In vitro, we found that OMCTS could promote the proliferation and migration of HTR8/SVneo cells. The model of combining clinical factors and OMCTS had a good performance in PAS prediction (AUC = 0.97, 95% CI 0.78-0.93). The early diagnosis of PAS in pregnant women requires assessing risk factors, metabolic status, and BS0 levels before 20weeks of gestation. OMCTS may be related to the development of PAS by promoting trophoblast cell proliferation and migration.

PUM1 modulates trophoblast cell proliferation and migration through LRP6.

Preeclampsia is a severe pregnancy complication characterized by hypertension and may cause maternal morbidity and mortality. A better understanding of the essential genes involved in preeclampsia pathophysiology is urgently needed. This study investigated the function and molecular mechanisms of pumilio RNA binding family member 1 (PUM1) in extravillous trophoblast cells (EVTs). The interaction between protein and mRNA was verified by RNA pull-down assays, RNA immunoprecipitation assays, and luciferase reporter assays. The mRNA and protein levels of the genes involved were determined by RT-qPCR and western blot assays, respectively. Our results demonstrated that PUM1 could bind to the 3'-untranslated region of low-density lipoprotein receptor-related protein 6 (LRP6) mRNA, resulting in reduced expression of LRP6 mRNA and protein. Repression of PUM1 resulted in enhanced colony formation, cell proliferation, migration, and invasion of EVTs. The PUM1-depletion-mediated promotion effects on EVTs could be abrogated by LRP6 knockdown. PUM1 regulates the growth and mobility of EVTs by modulating LRP6 expression. Developing strategies to balance PUM1 and LRP6 levels may be beneficial for the management of preeclampsia patients.

Porcine uterine luminal fluid-derived extracellular vesicles improve conceptus-endometrial interaction during implantation

Successful implantation of porcine conceptus requires synergistic interaction with various signal molecules in the maternal uterus. Extracellular vesicles (EVs) in uterine luminal fluid (ULF) of mice play important roles in conceptus development. However, studies have not explored the roles of extracellular vesicles (EV) in ULF of pigs. The aim of this study was to identify characteristics, origin, and roles of ULF-derived EVs on day 9 of the estrous cycle and on day 9,12 and 15 of pregnancy in pigs. Western blot, BCA assay and HE staining analysis showed increase in EVs concentration in ULF began from day 12 of pregnancy. Immunofluorescence staining and transmission electron microscopy analysis showed that EVs were mainly derived from endometrial epithelial cells. Fluorescent labeling, CCK-8 and transwell migration assays showed that these EVs were delivered to the trophoblast or parthenogenetic activation embryos to regulate proliferation and migration of trophoblast cells. A total of 305 miRNAs were identified using small RNA sequencing analysis. Functional enrichment analysis showed that miRNAs in these EVs potentially play vital regulatory functions in EV transportation or conceptus implantation. QRT-PCR analysis was used to further verify the RNA-seq data. The findings of this study provide information on the functions of porcine ULF-derived EVs and provide a reference dataset for future translational studies on porcine ULF-derived EVs.

Increased LINC00922 in preeclampsia regulates the proliferation, invasion, and migration of placental trophoblast cells.

BackgroundRecent studies have shown that the abnormal expression of long-chain non-coding RNAs (lncRNAs) can significantly affect the biological function of trophoblast cells and lead to the occurrence of preeclampsia (PE). This study explores the expression of lncRNA LINC00922 in PE and its effect on the function of placental trophoblast cells, along with the corresponding molecular mechanism, providing a theoretical basis and molecular target for understanding the occurrence, early diagnosis, and targeted therapy of PE.MethodsFluorescence quantitative PCR was used to detect the expression of LINC00922 in 30 cases of PE tissues and normal tissues. The CCK-8 assay, clone formation experiment, and flow cytometry were used to detect the effects of LINC00922 knockdown or overexpression on the proliferation, colony formation, and cell cycle of HTR-8/SVneo placental trophoblast cells. The Transwell assay was used to detect the effects of LINC00922 knockdown or overexpression on the invasion and migration of HTR 8/SVneo cells, and western blot was used to detect the expression of cell cycle-related proteins and invasion and migration-related proteins.ResultsLINC00922 was highly expressed in PE tissues. Knockdown of LINC00922 significantly inhibited the proliferation, invasion, and migration of HTR-8/SVneo cells, along with colony formation and the ability to induce cell cycle arrest in the G0/G1 phase. However, overexpression of LINC00922 had the opposite effect. Knockdown or overexpression of LINC00922 significantly affected the expression of cell cycle-related proteins cyclin-dependent kinase 2 (CDK2), G1/S-specific cyclin-D1 (Cyclin D1), p21, proliferating cell nuclear antigen (PCNA), matrix metallopeptidase 9 (MMP-9), vimentin, and E-cadherin, but had no significant effect on the expression of matrix metallopeptidase 2 (MMP-2).ConclusionsLINC00922 was highly expressed in PE, and functional experiments showed that LINC00922 could significantly affect the proliferation and invasion abilities of placental trophoblast cells, suggesting that LINC00922 may play an important role in the occurrence, early diagnosis, and treatment of PE.

Circular RNA hsa_circ_0026552 inhibits the proliferation, migration and invasion of trophoblast cells via themiR‑331‑3p/TGF‑βR1 axis in pre‑eclampsia.

Globally, pre-eclampsia (PE) is a gestational disorder that causes increased morbidity of the fetus and mortality induced by pregnancy. Despite various studies, the understanding of the causes or mechanism of the development of PE remains elusive. Thus, the present study aimed to investigate the role of circular (circ)RNA hsa_circ_0026552 (hsa_circ_0026552) in the development of PE and its mechanism of regulation. hsa_circ_0026552 differential expression in PE tissue data and clinical samples were analyzed and it was observed that hsa_circ_0026552 is highly upregulated in PE samples. Furthermore, miR-331-3p was detected as an hsa_circ_0026552 target miRNA and TGF-βR1 gene as a target of miR-331-3p. These results were confirmed using various assays, including dual-luciferase reporter, reverse transcription-quantitative PCR and RNA pull-down assay. It was observed that miR-331-3p expression was negatively correlated to hsa_circ_0026552 relative expression, while TGF-βR1 expression was positively correlated to hsa_circ_0026552 expression evaluated by Pearson's correlation test. The functional experiments, including Cell Counting Kit-8, colony formation and Transwell assay, showed that silencing hsa_circ_0026552 could significantly strengthen the proliferation, migration and invasion of the trophoblastic HTR-8/SVneo cells, but the subsequent overexpression of hsa_circ_0026552 reversed this. Mechanistically, it was concluded that hsa_circ_0026552 acts as a miR-331-3p sponge to upregulate TGF-βR1 expression in trophoblasts and is involved significantly in PE development and progression in pregnant women. The circRNA hsa_circ_0026552 could be a novel therapeutic target and prognostic biomarker for PE.

MiR-146a-5p-mediated suppression on trophoblast cell progression and epithelial-mesenchymal transition in preeclampsia

ObjectiveThis study aims to identify the effect of miR-146a-5p on trophoblast cell invasion as well as the mechanism in preeclampsia (PE).MethodsExpression levels of miR-146a-5p and Wnt2 in preeclamptic and normal placentae were quantified. Trophoblast cells (HTR-8) were separately transfected with miR-146a-5p mimic, miR-146a-5p inhibitor, pcDNA3.1-Wnt2 or sh-Wnt2, and then the expression levels of miR-146a-5p, Wnt2, and epithelial-mesenchymal transition (EMT)-related proteins (Vimentin, N-cadherin and E-cadherin) were measured. Moreover, the proliferative, migratory and invasive capacities of trophoblast cells were detected, respectively. Dual luciferase reporter assay determined the binding of miR-146a-5p and Wnt2.ResultsCompared with normal placental tissues, the placentae from PE patients showed higher miR-146a-5p expression and lower Wnt2 expression. Transfection of miR-146a-5p inhibitor or pcDNA3.1-Wnt2 exerted pro-migratory and pro-invasive effects on HTR-8 cells and encouraged EMT in HTR-8 cells; transfection with miR-146a-5p mimic or sh-Wnt2 weakened the proliferative, migratory and invasive capacities as well as reduced EMT process of HTR-8 cells. Moreover, Wnt2 overexpression could partially counteract the suppressive effects of miR-146a-5p overexpression on the progression and EMT of HTR-8 cells.ConclusionmiR-146a-5p mediates trophoblast cell proliferation and invasion through regulating Wnt2 expression.

Circular RNA circ_0111277 Serves as ceRNA, Targeting the miR-424-5p/NFAT5 Axis to Regulate the Proliferation, Migration, and Invasion of Trophoblast Cells in Preeclampsia.

Preeclampsia is the main reason for maternal and fetal deaths during the second half of pregnancy. Trophoblast cells play a pivotal role in preeclampsia progression. Circular RNA (circRNA) circ_0111277 has been reported to be related to the development of trophoblast cells. This study is designed to explore the role and mechanism of circ_0111277 on trophoblast cell behavior in preeclampsia. Circ_0111277, microRNA-424-5p (miR-424-5p), and nuclear factor of activated T-cell 5 (NFAT5) levels were measured by real-time quantitative polymerase chain reaction (RT-qPCR). Cell viability, migration, invasion, and angiogenesis were measured by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay, transwell assay, tube formation assay, and wound healing assay. Protein levels of matrix metallopeptidase 2 (MMP2), vascular endothelial growth factor-A (VEGF-A), NFAT5, phospho-phosphatidylinositol 3 kinase (p-PI3K), PI3K, phospho-protein kinase B (p-AKT), and AKT were examined by western blot assay. The binding relationship between miR-424-5p and circ_0111277 or NFAT5 was predicted by circBank or starBase and then verified by a dual-luciferase reporter assay. Circ_0111277 and NFAT5 expression were increased in placenta tissues of preeclampsia patients, and miR-424-5p was decreased. Moreover, circ_0111277 knockdown could boost cell viability, migration, invasion, and angiogenesis in trophoblast cells. The mechanical analysis discovered that circ_0111277 acted as a sponge of miR-424-5p to regulate NFAT5 expression. Besides, circ_0111277 silencing promoted the PI3K/AKT signaling pathway in trophoblast cells. Circ_0111277 downregulation could facilitate cell growth and metastasis in trophoblast cells partly by regulating the miR-424-5p/NFAT5 axis, providing an underlying circRNA-targeted therapy for preeclampsia.

Circ_0007121 Facilitates Trophoblastic Cell Proliferation, Migration, and Invasion via the Regulation of the miR-421/ZEB1 Axis in Preeclampsia.

Noncoding circular RNAs (circRNAs) have participated in the progression of preeclampsia (PE) via inhibiting microRNAs (miRNAs) to regulate gene expression. This study was designed to explore the miRNA/mRNA mechanism of hsa_circ_0007121 (circ_0007121) in PE. The expression detection of circ_0007121, microRNA-421 (miR-421), and zinc finger E-box binding homeobox 1 (ZEB1) was performed by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was analyzed by Cell Counting Kit-8 (CCK-8) assay. Transwell assay was used to determine cell migration and invasion. Cell apoptosis was evaluated using flow cytometry. The protein levels of epithelial-mesenchymal transition (EMT) markers and ZEB1 were measured via western blot. The interaction between miR-421 and circ_0007121 or ZEB1 was validated by dual-luciferase reporter assay. The expression detection indicated that circ_0007121 was downregulated in PE patients and the clinical data revealed that circ_0007121 was related to PE. The upregulation of circ_0007121 promoted cell proliferation, migration, invasion, and EMT in trophoblastic cells. Furthermore, circ_0007121 was identified as a sponge of miR-421 and the function of circ_0007121 was dependent on the sponge effect on miR-421. Moreover, ZEB1 was a target of miR-421 and circ_0007121/miR-421 axis could regulate the expression of ZEB1. In addition, miR-421 overexpression repressed trophoblastic cell behaviors through downregulating the ZEB1 level. Altogether, circ_0007121 contributed to the development of trophoblastic cells by regulating the miR-421/ZEB1 axis.