Gao Tian, Deng Mou, Wang Qian
Department of Gynecology and Obstetrics, the First Affiliated Hospital, Chongqing Medical University, Chongqing, China.
J Obstet Gynaecol Res. 2018 Apr;44(4):756-763. doi: 10.1111/jog.13560. Epub 2018 Jan 18.
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.
微小RNA（miR）在滋养层细胞功能调节中起重要作用。我们探究了miR-320a在人胎盘中的表达。此外，我们报告了miR-320a在滋养层细胞中的显著作用及靶功能环。
通过定量实时聚合酶链反应研究子痫前期和健康胎盘组织中miR-320a的表达，以确定miR-320a如何影响滋养层细胞的侵袭、增殖和迁移。在滋养层细胞中建立脂多糖（LPS）模型，以揭示补充LPS如何刺激miR-320a的表达。应用蛋白质免疫印迹法测量蛋白质表达，该蛋白质参与子痫前期胎盘中由miR-320a调节的信号通路。
子痫前期患者胎盘标本中miR-320a表达增强。过量的miR-320a表达显著抑制滋养层细胞侵袭，但不影响迁移或增殖。然而，用miR-320a抑制剂转染可增强体外滋养层细胞的侵袭能力。荧光素酶报告基因检测证实雌激素相关受体γ（ERRγ）是miR-320a的直接靶点。定量实时聚合酶链反应和蛋白质免疫印迹法表明过量的miR-320a表达下调了ERRγ转录和翻译。此外，补充LPS显示miR-320a表达过量和ERRγ下调。与对照组相比，子痫前期胎盘中ERRγ受损且miR-320a表达增强。Pearson相关性和线性回归分析显示，在正常和子痫前期胎盘中，miR-320a表达与ERRγ表达呈负相关。
这些发现表明miR-320a过表达通过靶向ERRγ导致胎盘形成异常。我们的研究揭示了miR-320a和ERRγ功能环对胎盘形成的显著作用。
