miR-15b-AGO2 play a critical role in HTR8/SVneo invasion and in a model of angiogenesis defects related to inflammation.

microRNAs (miRs) have been shown to play critical roles in the regulation of trophoblast and endothelial cell functions, and one significant finding concerning the miR-15/16 family is that most members of this family are highly expressed in endothelial cells and contribute to functions, such as tube formation. The interaction between trophoblast and endothelial cell play an important role in normal placentation process. Therefore, the aims of this study were to investigate the expression of miR-15b in human placenta and to uncover the potential role of miR-15b as well as its target functional loop in trophoblast and endothelial cells. Whether inflammation could modulate the expression of miR-15b and its down-stream target was further investigated. Additionally, the potential link between miR-15b deregulation and preeclampsia was also explored in the placenta of patients diagnosed with preeclampsia. The expression of miR-15b was studied in the placental tissue of a normal pregnancy using in situ hybridization, and the effects of miR-15b on proliferation, invasion, and angiogenesis were further explored in vitro using HTR-8/SVneo and HUVEC cell line models. A Lipopolysaccharides (LPS) treatment model in HTR-8/SVneo cell was utilized to explore the mechanism of how LPS treatment could lead to the activation of miR-15b expression. Western blot was used to detect the expression of proteins related to miR-15b mediated pathway in preeclamptic placentas. miR-15b inhibits trophoblast cell invasion and endothelial cell tube formation by suppressing the expression of Argonaute 2 (AGO2), a major miRNA effecter protein. AGO2 is specifically localized to human placenta cytotrophoblast and endothelial cells, and it plays important roles in trophoblast cell invasion and endothelial cell tube formation. LPS treatment may lead to the overexpression of miR-15b and down-regulation of AGO2, which may be involved in shallow trophoblast cell invasion associated with the pathogenesis of preeclampsia. Chromatin immunoprecipitation assay indicates that increased occupancy of AGO2 to miR-15b promoter is responsible for the increased expression of miR-15b under the condition of LPS treatment. Furthermore, preeclamptic placentas have decreased expression of AGO2, but increased expression of miR-15b and TLR-4 compared to normal controls. This is the first report about the function of AGO2 in human trophoblast and endothelial cells in the placenta. The data indicates that the aberrant expression of miR-15b contributes to abnormal placentation by targeting AGO2 mRNA. This study provides insight into the potential role of the miR-15b and AGO2 functional loop in the placentation process.

Yang M ，Chen Y ，Chen L ，Wang K ，Pan T ，Liu X ，Xu W ... -  《-》

MicroRNA-210 regulates human trophoblast cell line HTR-8/SVneo function by attenuating Notch1 expression: Implications for the role of microRNA-210 in pre-eclampsia.

Successful pregnancy depends on the precise regulation of extravillous trophoblast cell invasion ability. MicroRNA-210-3p (miR-210), which is increased in the placenta of pre-eclampsia. Furthermore, miR-210 could inhibit trophoblasts invasion and might act as a serum biomarker for pre-eclampsia. Previous studies have demonstrated that miR-210 regulates HUVEC (human umbilical vein endothelial cell)-mediated angiogenesis by regulating the NOTCH1 signaling pathway. Studies by our group have previously identified that NOTCH1 plays a positive role in regulating trophoblast functions. However, the miR-210/NOTCH1 signaling pathway in the regulation of trophoblasts and pre-eclampsia has not been characterized. Therefore, this study was conducted to investigate the role of miR-210 and its relationship with NOTCH1 in trophoblasts. We first examined the expression levels of miR-210 and NOTCH1 in pre-eclamptic and normals placentas. Next, the expression and location of miR-210 and NOTCH1 in the first-trimester villi, maternal decidua, and placenta of late pregnancy were shown via in situ hybridization and immunohistochemistry. The trophoblast cell line HTR-8/SVneo was used to investigate the effects of miR-210 on the expression of NOTCH1 and cell bioactivity by upregulation and downregulation strategies. The results showed that miR-210 expression was increased, whereas NOTCH1 expression was decreased in pre-eclamptic placenta compared with controls. Upregulation of miR-210 decreased NOTCH1 expression, impaired HTR-8/SVneo proliferation, migration, invasion, and tube-like formation capabilities, and promoted apoptosis. In contrast, downregulation of miR-210 resulted in the opposite effects. These findings suggested that miR-210 might act as a contributor to trophoblast dysfunction by attenuating NOTCH1 expression.

Wang R ，Liu W ，Liu X ，Liu X ，Tao H ，Wu D ，Zhao Y ，Zou L ... -  《-》

MiR-195 participates in the placental disorder of preeclampsia via targeting activin receptor type-2B in trophoblastic cells.

Wu H ，Wang H ，Liu M ，Bai Y ，Li YX ，Ji L ，Peng C ，Yu Y ，Wang YL ... -  《-》

Wnt5a inhibited human trophoblast cell line HTR8/SVneo invasion: implications for early placentation and preeclampsia.

Chen Y ，Zhang Y ，Deng Q ，Shan N ，Peng W ，Luo X ，Zhang H ，Baker PN ，Tong C ，Qi H ... -  《-》

MiRNA-320a inhibits trophoblast cell invasion by targeting estrogen-related receptor-gamma.

MicroRNAs (miRs) play an essential role in the modulation of trophoblast function. We explored miR-320a expression in the human placenta. In addition, we report the promising effect and target functional loop of miR-320a in trophoblasts. MiR-320a expression was investigated in both pre-eclamptic and healthy placenta tissues by quantitative real time-polymerase chain reaction to determine how miR-320a affected invasion, proliferation and migration in trophoblasts. A lipopolysaccharide (LPS) model was established in trophoblasts to reveal how LPS supplementation stimulated miR-320a expression. Western blot was applied to measure protein expression, which was involved in pathways modulated by miR-320a in pre-eclamptic placentas. MiR-320a expression was enhanced in the placental specimens of pre-eclamptic patients. Excessive miR-320a expression remarkably suppressed trophoblast invasion but did not affect migration or proliferation. However, transfection with miR-320a inhibitor reinforced trophoblast invasion in vitro. Luciferase assays verified that estrogen-related receptor-gamma (ERRγ) served as a direct target of miR-320a. Quantitative real-time polymerase chain reaction and Western blot demonstrated that excessive miR-320a expression downregulated ERRγ transcription and translation. Additionally, LPS supplementation showed excessive miR-320a expression and ERRγ downregulation. Impaired ERRγ and enhanced miR-320a expression occurred in PE placentas compared to controls. Pearson correlation and linear regression analysis revealed that miR-320a expression was negatively related to ERRγ expression in normal and pre-eclamptic placentas. These findings indicate that miR-320a overexpression causes anomalous placentation by targeting ERRγ. Our research reveals the promising effect of miR-320a and the ERRγ functional loop on placentation.

Gao T ，Deng M ，Wang Q 《-》