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MicroRNA-141 (miR-141-3p) is upregulated in preeclampsia. This study investigated the effect of methylation of the miR-141-3p promoter on cell viability, invasion capability, and inflammasomes in vitro. The expression of miR-141-3p and methylation status of the miR-141-3p promoter were examined by RT-qPCR and pyrosequencing in villus tissues of women with spontaneous delivery (VTsd), villus tissues of women with preeclampsia (VTpe), and also in HTR-8/SVneo cells treated with a miR-141-3p inhibitor and 20 μmol/L 5-aza-2′-deoxycytidine (5-Aza), a DNA methyltransferase inhibitor. Cell viability and invasion were evaluated by CCK-8 and transwell assays. In addition, the levels of CXCL12, CXCR4, CXCR2, MMPs, NLRP3, and ASC expression were assessed by western blotting, and IL-1β and IL-18 concentrations were assayed by ELISA. miR-141-3p expression was upregulated, and the levels of miR-141-3p promoter methylation and CXCL12, CXCR4, and CXCR2 expression were decreased in VTpe relative to VTsd. In HTR-8/SVneo cells, hypomethylation caused by 5-Aza treatment increased miR-141-3p expression, while DNA methyltransferase 3 (DNMT3) transfection decreased miR-141-3p expression. miRNA-141-3p induced NLRP3, IL-1β, and IL-18 production, decreased CXCR4, MMP, and MMP2 production, and suppressed cell growth and invasion. Furthermore, we observed that NLRP3 plays an important mediatory role in the effects of miR-141-3p described above. Decreased methylation of the miR-141-3p promoter increases miR-141-3p expression, which in turn increases NLRP3 expression, resulting in higher IL-1β and IL-18 levels and lower levels of MMP2/9 and CXCR4. We conclude that modification of the miR-141-3p promoter might be a curial mediator in preeclampsia.

Background MiR-135a-5p is an important regulator of cell migration and invasion in several diseases. However, the biological functions and mechanisms of miR-135a-5p in women with preeclampsia (PE) remain unclear. Methods The levels of miR-135a-5p and beta-transducin repeat containing E3 ubiquitin protein ligase (β-TrCP) expression in samples of placenta tissue from PE patients and healthy control subjects were determined by quantitative real-time PCR. The effects of miR-135a-5p and β-TrCP on cell migration, invasion, and epithelial-mesenchymal transition (EMT) in two trophoblast cell lines (HTR-8/SVneo and TEV-1) were examined using wound healing, Transwell, and western blot assays, respectively. A luciferase reporter assay was performed to confirm the association between miR-135a-5p and β-TrCP, and an in vivo mouse model was established and used to analyze the effect of β-TrCP on PE clinical phenotypes. Results We found that miR-135a-5p expression was significantly decreased and negatively correlated with β-TrCP expression in the placental tissues of pregnant women with PE. Cellular function experiments showed that overexpression of miR-135a5p promoted the migration and invasion of trophoblast cells in vitro. Furthermore, β-TrCP was confirmed as a target gene of miR-135a-5p in trophoblast cells. Notably, overexpression of β-TrCP significantly reversed the effect of miR-135a-5p on migration and invasion of trophoblast cells. At the molecular level, decreases in E-cadherin levels and increases in N-cadherin, Vimentin, and β-catenin levels that were induced by miR-135a-5p overexpression were attenuated by β-TrCP overexpression. Conclusions Our findings demonstrate that miR-135a-5p promotes the migration and invasion of trophoblast cells by targeting β-TrCP.

Endometrial cancer, one of the most common gynecological cancers in women. Patients with advanced or recurrent disease have poor long-term outcomes. The current experiment explore the roles of cationic microbubbles (CMBs) carrying paclitaxel (PTX) and CRISPR/Cas9 plasmids on the xenotransplantation model of mice with endometrial cancer. The tumor histology, tumor cell viability, cell cycle, and invasion ability were investigated. Meanwhile, the P27, P21, GSK-3, Bcl-2 associated death promoter (Bad), mammalian target of rapamycin (mTOR), and C-erbB-2 expressions were evaluated by qRT-PCR and western blotting, respectively. CMB-PTX-CRISPR/Cas9 had an inhibitory action on the tumor growth, tumor cell viability, cell cycle, and invasion ability of the mouse xenograft model of endometrial cancer. The CMB-PTX-CRISPR/Cas9 increased the GSK-3, P21, P27, and Bad expression levels, while reduced the C-erbB-2 and mTOR expressions. CMBs loaded with both PTX and CRISPR/Cas9 plasmids may be a new combination treatment with much potential. CMB-PTX-CRISPR/Cas9 may regulate the tumor cell viability, invasion, and metastasis of endometrial cancer naked mouse model by upregulating expressions of GSK-3, P21, P27, and Bad.

Preeclampsia is a pregnancy disorder that is primarily associated with maternal and neonatal or fetal morbidity and mortality. The discovery of dysregulated microRNAs (miRs) and their roles in preeclampsia has provided new insight into the mechanisms involved in pregnancy-related disorders. In the present study, quantitative PCR demonstrated that the expression levels of miR-524-5p were lower in patients with preeclampsia than those in normal pregnant women. Cell Counting Kit-8 and Transwell assays indicated that overexpression of miR-524-5p promoted the proliferation and invasion of HTR-8/SVneo cells, whereas inhibition of miR-524-5p suppressed HTR-8/SVneo cell proliferation and invasion. Furthermore, NUMB endocytic adaptor protein (NUMB), a negative regulator of the Notch signaling pathway and a target gene of miR-524-5p, limited the effects of miR-524-5p on HTR-8/SVneo cell invasion and migration. The present study demonstrated that miR-524-5p regulated the proliferation and invasion of HTR-8/SVneo cells at least partly by targeting NUMB to regulate the Notch signaling pathway.

Insufficient trophoblast invasion has been shown to contribute to the occurrence and progression of preeclampsia (PE). Recently, beta-transducin repeat containing E3 ubiquitin protein (β-TrCP) was shown to function as a ubiquitination regulator in regulating the proliferation and invasion of various cell types. In this study, we employed an in vitro model of trophoblasts to investigate the role played by β-TrCP in the pathogenesis of PE. The levels of β-TrCP in newly delivered placentas from 15 pregnant women with PE and 15 healthy pregnant women were detected by quantitative real-time PCR and western blot assays. The effects of β-TrCP on cell migration, invasion, and epithelial-mesenchymal transition (EMT) in two trophoblast cell lines (HTR-8/SVneo and TEV-1) were examined using wound healing assays, Transwell assays, and western blot assays, respectively. Rescue experiments were performed by treating β-TrCP knockdown or β-TrCP expressing trophoblasts with si-Snail transfection or a proteasome inhibitor (MG132). β-TrCP mRNA and protein expression levels were significantly increased in the PE placentas when compared to the normal control placentas. β-TrCP overexpression significantly inhibited cell migration and invasion, while silencing of β-TrCP promoted cell migration and invasion of the two trophoblast cell lines. Furthermore, we demonstrated that β-TrCP-mediated ubiquitination might inhibit the EMT process of trophoblasts by down-regulating Snail expression. Our results suggest that both β-TrCP mRNA and protein expression were up-regulated in the PE placentas. β-TrCP impeded the migration and invasion of trophoblasts by suppressing Snail expression. This implicates the ubiquitin-proteasome pathway in the pathogenesis of PE, and suggests β-TrCP as a potential target for treating PE.

Objectives: To investigate the relationship between hsa_circ_0051326 and HLA-G expression in the blood of patients with pre-eclampsia. Material and methods: Real-time PCR (qRT-PCR) was used to detect the hsa_circ_0051326 expression level. Enzyme-linked immunosorbent assay (ELISA), qRT-PCR, and western blotting were used to detect HLA-G expression in blood in 50 patients with pre-eclampsia and 50 normal pregnant women. Results: HLA-G protein expression level was decreased significantly in the blood of patients with pre-eclampsia. hsa_circ_0051326 and HLA-G mRNA in blood were decreased significantly in the pre-eclampsia patients compared with normal pregnant women. There was a positive correlation between the expression of serum hsa_circ_0051326 with HLA-G mRNA. Conclusions: Serum hsa_circ_0051326 was a new diagnostic biomarker for pre-eclampsia with equivalent efficiency of HLA-G. Maternal serum hsa_circ_0051326 level may pre-diagnose potentially pre-eclampsia before delivery.

Background/Aims: Cancer stem-like cells are the main cause of tumor occurrence, progression, and therapeutic resistance. However, the precise signals required for the maintenance of the stem-like traits of these cells in ovarian cancer remain elusive. We have thus worked to elucidate the functional role of Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ), a gene encoding the 14-3-3ζ protein, in the regulation of multidrug resistance and stem cell-like traits in ovarian cancer. Methods: We detected the YWHAZ levels in human ovarian cancer specimens and cell lines using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blots. MTS assays, soft agar colony formation assays, migration assays, cell cycle analysis, sphere formation assays, and flow cytometry were applied to investigate the functional role of YWHAZ in ovarian cancer. Results: Our data reveals substantially increased YWHAZ expression in both cisplatin- and paclitaxel-resistant ovarian cancer cells. Silencing YWHAZ restored the sensitivity of resistant ovarian cancer cells to cisplatin and paclitaxel. Furthermore, in vitro studies showed that down-regulation of YWHAZ inhibited cell cycle progression, migration, and the expression of stem cell markers. Moreover, tumorigenicity was suppressed in tumor-bearing BALB/c nude mice following YWHAZ knockdown. Additionally, we demonstrated that the expression of YWHAZ was directly down-regulated by miR-30e in resistant ovarian cancer cells. Conclusion: Our results have led to new insights into the essential role of YWHAZ in the regulation of tumourigenesis, stem-like traits, and drug resistance in ovarian cancer, thereby helping to identify a potential target for ovarian cancer therapy.

Background An impaired trophoblast invasion ability contributes to the development of pre-eclampsia (PE), and can be induced by the altered expression of various microRNAs (miRs). MiR-141 and CXCL12β (C-X-C motif chemokine ligand 12) signaling regulate trophoblast invasion and vascularization capabilities during PE pathogenesis; however, their interactions and underlying mechanisms of action remain unclear. We investigated how miR-141 modulates trophoblast invasion, with a focus on its interaction with CXCL12β signaling. Methods A PE model was established by using HTR-8/SVneo cells, which were first cultured with 2% O2 for 48 h, and then with 5% O2. The expression of miR-141 in human villous trophoblast HTR-8/SVneo cells was modulated with mimics or an inhibitor, and analyzed by quantitative RT-PCR. CXCL12β levels were determined by ELISA. Cell apoptosis was determined by flow cytometry, and the invasion and vascularization capabilities of trophoblasts were evaluated by Transwell and tube formation assays, respectively. Binding of miR-141 with CXCL12β mRNA was verified by the dual luciferase assay. Protein levels were estimated by western blotting. Results MiR-141 expression was significantly induced by hypoxia in HTR-8/SVneo cells. MiR-141 was found to promote apoptosis and inhibit the invasion and vascularization abilities of HTR-8/SVneo cells under conditions of hypoxia. MiR-141 could directly bind with the 3′UTR region of CXCL12β mRNA and inhibit its translation. In addition, we proved that miR-141 could inhibit the invasion and vascularization abilities, and promote the apoptosis of HTR-8/SVneo cells by targeting CXCL12β under hypoxic conditions. Furthermore, we demonstrated that arachidonic acid could reverse the invasion and apoptosis abilities of HTR-8/SVneo cells mediated by CXCL12β during hypoxia. In terms of mechanism, MiR-141 could downregulate MMP2, p62, and LC3B expression, and upregulate ROCK1 and RhoA expression in HTR-8/SVneo cells by targeting the CXCL12β gene during hypoxia. The effects of CXCL12βon HTR-8/SVneo cells could be reversed by arachidonic acid (ARA). Conclusion Induction of miR-141 by hypoxia promotes apoptosis, and inhibits the invasion and vascularization capabilities of HTR-8/SVneo cells by suppressing the CXCL12β and CXCR2/4 signaling pathways.

It has been well established that dietary patterns play important roles in the pathogenesis and development of hypertension. Our aim was to investigate the association between pregnancy dietary patterns and the risk of hypertension among nulliparous pregnant Chinese women. A cross-sectional, case-control study. Three hospitals in Haikou, the capital of Hainan Province, South China. A total of 2580 participants who reported dietary intake using a validated food frequency questionnaire (FFQ). Four primary dietary patterns were identified by principal component factor analysis and labeled as traditional Chinese, animal food, Western food, and salty snacks patterns. Women with high scores on pattern characterized by salty snacks were at increased risk. This study suggests that dietary pattern characterized by salty snack increases the risk of hypertension during pregnancy.

Introduction CXCR2, the receptor of the CXC chemokines, plays a critical role in cell migration and invasion in many types of cancer. It is unclear what impact CXCR2 may have on Preeclampsia (PE), a pregnancy-specific disease, which is related to insufficient trophoblast invasion. The aim of this study was to investigate the expression pattern of CXCR2 in the placentas of healthy and PE pregnancies, and to investigate the molecular mechanism of CXCR2 involvement in the development of PE. Methods CXCR2 expression levels in newly delivered placentas from 38 pregnant women with PE and 21 healthy pregnant women were detected using quantitative real-time PCR, immunohistochemistry and Western blot assays. The effect of CXCR2 on trophoblast invasion and the underlying mechanisms were examined in two trophoblast cell lines (HTR-8/SVneo and TEV-1 cells). Results CXCR2 mRNA and protein expression levels were significantly decreased in preeclamptic placentas than normal control. The invasive abilities of the two trophoblast cell lines were significantly inhibited when CXCR2 was silenced, but that CXCR2 overexpression promoted trophoblast cells invasion. In addition, silencing CXCR2 reduced the expression of matrix metalloproteinase 2 and 9 (MMP2 and MMP9) and phosphorylated Akt (p-Akt). Furthermore, an Akt inhibitor suppressed the expression of MMP-2 and MMP-9. Discussion Our results suggest that the decreased CXCR2 may contribute to the development of preeclampsia through impairing trophoblast invasion by down-regulating MMP-2 and MMP-9 via the Akt signaling pathway.

To identify the association between preeclampsia (PE) and selective intrauterine growth restriction (sIUGR) in twin pregnancies.This was a retrospective cohort study of 1004 twin pregnancies from 2008 to 2014. We specifically compared the incidence, clinical characteristics and outcomes of PE between sIUGR and normal-growth twin pregnancies.PE occurred more frequently in sIUGR pregnancies [29.0% (51/176)] than in normal-growth twin pregnancies [13.1% (99/756), p 0.001, adjusted odds ratio 3.29]. Among sIUGR, the incidence of PE was significantly higher in dichorionic (DC) pregnancies (37.5%, 30/80) than in monochorionic (MC) pregnancies (21.9%, 21/96). The rates of onset at32 weeks (p = 0.045) and of severe PE (p = 0.025) were higher in sIUGR pregnancies with PE. The systolic blood pressure was also higher in sIUGR pregnancies with PE (152.6 ± 11.8 mmHg) than in normal-growth pregnancies with PE (148.0 ± 8.2 mmHg) (p = 0.042). Additionally, more sIUGR pregnancies were delivered at 32-36 weeks (p = 0.001), and fewer were delivered at ≥36 weeks (p 0.001). Moreover, the prevalence of severe neonatal asphyxia was higher in sIUGR pregnancies with PE than in normal-growth pregnancies with PE (8.8% versus 2.5%, p = 0.020).sIUGR is associated with increased odds of developing severe PE in twin pregnancies, leading to poorer perinatal outcomes.