miR-183-5p suppressed the invasion and migration of HTR-8/SVneo trophoblast cells partly via targeting MMP-9 in preeclampsia.

Preeclampsia (PE), a common obstetrical disorder, is characterized by impaired migration and invasion abilities of trophoblastic cells. MicroRNA-183-5p (miR-183) was reported to regulate cell migration and invasion in various types of human cancers; however, its role in the pathogenesis of PE remains elusive. Herein, we investigated the role of miR-183 in HTR-8/SVneo trophoblast cells invasion and migration and explored the underlying mechanism. Our results showed that miR-183 was significantly up-regulated in placental tissues from pregnant women compared with that in normal pregnant women. Overexpression of miR-183 inhibited proliferation, migration and invasion, as well as induced apoptosis in HTR-8/SVneo cells. Otherwise, down-regulation of miR-183 achieved the opposite effects. Bioinformatics prediction and luciferase reporter assay confirmed that matrix metalloproteinase-9 (MMP-9) is a target of miR-183. In addition, MMP-9 expression was significantly down-regulated, and inversely correlated with the miR-183 level in placental tissues from pregnant women with severe PE. Down-regulation of MMP-9 suppressed the trophoblast cell invasion and migration, whereas overexpression of MMP-9 promoted cell invasion and migration in HTR-8/SVneo cells. More importantly, up-regulation of MMP-9 reversed the inhibitory effects of miR-183 on cell invasion and migration in trophoblast cells. Collectively, our findings suggested that miR-183 may play critical roles in the pathogenesis of PE and serve as a potential biomarker for severe PE.

Suo M ，Sun Y ，Yang H ，Ji J ，He Y ，Dong L ，Wang Y ，Zhang Y ，Zhang Y ，Hao M ... -  《-》

Up-regulation of miR-299 suppressed the invasion and migration of HTR-8/SVneo trophoblast cells partly via targeting HDAC2 in pre-eclampsia.

Pre-eclampsia (PE), a pregnancy-associated disorder, is a major contributor to maternal mortality and morbidity worldwide. Recently, microRNAs (miRNAs) were found to be associated with the pathogenesis of PE. The present study investigated the function of miR-299 in HTR-8/SVneo trophoblast cells and explored its underlying mechanism in the pathogenesis of PE. The miR-299 and histone deacetylase 2 (HDAC2) mRNA expression levels were determined by quantitative real-time PCR. Transwell invasion and wound healing assays were used to measure cell invasive and migratory ability. Luciferase reporter assay was performed to confirm the downstream targets of miR-299. Western blot was performed to measure the protein expression of HDAC2. The expression level of miR-299 in placental tissues from pregnant women with severe PE was significantly higher than that from normal pregnant women. MiR-299 overexpression suppressed the invasion and migration of trophoblast cells; and knock-down of miR-299 promoted the invasion and migration of trophoblast cells. Bioinformatics prediction and luciferase reporter assay confirmed that miR-299 directly targeted the 3' untranslated region of HDAC2. The mRNA expression level of HDAC2 in placental tissues from pregnant women with severe PE was significantly lower than that from normal pregnant women, and was negatively correlated the expression level of miR-299 in placental tissues from pregnant women with severe PE. HDAC2 siRNA transfection suppressed the mRNA and protein expression levels of HDAC in trophoblast cells compared with control group, and HDAC2 siRNA transfection also suppressed the trophoblast cell invasion and migration compared with control group. Enforced expression of HDAC2 in trophoblast cells attenuated the inhibitory effects of miR-299 overexpression on cell invasion and migration. In summary, the results showed the up-regulation of miR-299 in the placental tissues from women with severe PE and miR-299 suppressed the invasion and migration of trophoblast cells partly via targeting HDAC2.

Gao Y ，She R ，Wang Q ，Li Y ，Zhang H ... -  《-》

MiR-221-3p is down-regulated in preeclampsia and affects trophoblast growth, invasion and migration partly via targeting thrombospondin 2.

Preeclampsia (PE) is a common obstetrical disorder characterized by hypertension and proteinuria. The aberrant expression of miR-221-3p in placental tissues has been implicated in the pathogenesis of PE. This study examined the role of miR-221-3p in trophoblast growth, invasion and migration, and explored the underlying mechanisms. Our results showed that miR-221-3p was down-regulated in placental tissues from PE patients compared to heathy controls as measured by quantitative real-time PCR assay. CCK-8 assay, Transwell invasion assay and wound healing assay showed that miR-221-3p overexpression promoted trophoblast (HTR-8/SVneo cells) growth, invasion and migration, and knockdown of miR-221-3p exerted the opposite effects. Flow cytometry experiments further demonstrated that miR-221-3p overexpression inhibited apoptosis, increased cell population at S phase, and decreased cell population at G0/G1 phase of HTR-8/SVneo cells; while miR-221-3p knockdown exerted the opposite effects. Bioinformatics prediction and luciferase reporter assay showed that miR-221-3p targeted the 3' untranslated region of thrombospondin 2 (THBS2), and qRT-PCR and western blot assays revealed that miR-221-3p negatively regulated the expression of THBS2 in HTR-8/SVneo cells. Furthermore, overexpression of THBS2 attenuated the in vitro effects of miR-221-3p overexpression on cell growth, invasion and migration of HTR-8/SVneo cells. The clinical sample analysis showed that the mRNA expression levels of THBS2 was significantly increased in placental tissues from PE patients and was negatively correlated with expression levels of miR-221-3p. In summary, our results demonstrated that miR-221-3p was down-regulated in PE, and the in vitro functional assays revealed that miR-221-3p promoted trophoblast growth, invasion and migration partly via targeting THBS2.

Yang Y ，Li H ，Ma Y ，Zhu X ，Zhang S ，Li J ... -  《-》

Upregulated long noncoding RNA Linc00261 in pre-eclampsia and its effect on trophoblast invasion and migration via regulating miR-558/TIMP4 signaling pathway.

Pre-eclampsia (PE) is a leading cause of maternal and perinatal morbidity and mortality but the exact underlying mechanisms of PE pathogenesis remain elusive. Accumulated data suggested that the long noncoding RNAs (lncRNAs) play important roles in the pathogenesis of PE. The present study identified the changes of lncRNA Linc00261 in PE and its effects on trophoblasts invasion and migration. Our results showed that the expression of Linc00261 was upregulated in placental tissues of PE women compared with those of healthy pregnant women. Overexpression of Linc00261 suppressed cell invasion and migration, induced cell apoptosis, and caused cell-cycle arrest at G0 /G1 phase of HTR-8/SVneo cells; while knockdown of Linc00261 had the opposite effects on the HTR-8/SVneo cells. Mechanistic studies showed Linc00261 functioned as a competing endogenous RNA for miR-558 in HTR-8/SVneo cells, and miR-558 was negatively regulated by Linc00261. The expression level of miR-558 in the PE group was significantly lower than the control group, and the expression level of Linc00261 was negatively correlated with the expression level of miR-558 in the placental tissues of women with PE. Furthermore, miR-558 was found to negatively regulate the expression of TIMP metallopeptidase inhibitor 4 (TIMP4) via targeting the 3' untranslated region in the HTR-8/SVneo cells. Overexpression of miR-558 increased HTR-8/SVneo cell invasion and migration, which was attenuated by TIMP4 overexpression. More importantly, both overexpression of miR-558 and knockdown of TIMP4 partially reversed the suppressive effects of Linc00261 overexpression on cell invasion and migration of HTR-8/SVneo cells. Collectively, our results for the first time showed the upregulation of Linc00261 in the placental tissues of severe PE patients. The mechanistic results indicated that Linc00261 exerted the suppressive effects on the trophoblast invasion and migration via targeting miR-558/TIMP4 axis, which may involve in the pathogenesis of PE.

Cheng D ，Jiang S ，Chen J ，Li J ，Ao L ，Zhang Y ... -  《-》

Highly expressed miR-182-5p can promote preeclampsia progression by degrading RND3 and inhibiting HTR-8/SVneo cell invasion.

Fang YN ，Huang ZL ，Li H ，Tan WB ，Zhang QG ，Wang L ，Wu JL ... -  《-》