Zhu Qi, Dong Yi-Chao, Zhang Lu, Ma Xu, Xia Hong-Fei
Reproductive and Genetic Center of National Research Institute for Family Planning, Beijing, China.
Graduate Schools, Peking Union Medical College, Beijing, China.
Reproduction. 2020 May;159(5):525-537. doi: 10.1530/REP-19-0282.
Missed abortion (MA) is a common disease in obstetrics and gynecology. More and more studies have focused on the relationship between miRNAs and pregnancy maintenance and its related diseases. The aim of this article is to explore the relationship between miRNA and MA. The expression of miR-98 were detected by in situ hybridization and real-time PCR. Cell proliferation, activity and migration were measured via Edu, MTT, and transwell assays. The target genes of miR-98 are identified by dual-luciferase activity assay. And the expression levels of target genes were determined by Western blot, real-time PCR and immunohistochemistry. miR-98 was significantly up-regulated in placental villi from over 35 years old MA patients compared with the age-matched normal pregnant women. Up-regulation of miR-98 suppressed the proliferation, activity and migration of the human trophoblast HTR-8/SVneo cell in vitro. miR-98 could bind to GDF6 and FAPP2 mRNA 3'-UTR and negatively regulate their expression. The downregulation of miR-98 promoted cell proliferation, then knockdown of GDF6 or FAPP2 inhibited miR-98-mediated cell proliferation. GDF6 and FAPP2 expression in the placental villi from MA patients were decreased compared to normal placental tissues. The expression of miR-98 in MA had an opposite relationship with the expression of GDF6 and FAPP2. Overexpression of miR-98 is associated with the occurrence of MA. miR-98 prevents proliferation, viability and migration of trophoblast cells partially through targeting GDF6 and FAPP2.
稽留流产（MA）是妇产科的一种常见疾病。越来越多的研究聚焦于微小RNA（miRNAs）与妊娠维持及其相关疾病之间的关系。本文旨在探讨miRNA与稽留流产之间的关系。通过原位杂交和实时定量聚合酶链反应（PCR）检测miR-98的表达。采用EdU、MTT和Transwell实验检测细胞增殖、活性和迁移情况。通过双荧光素酶活性实验鉴定miR-98的靶基因。并通过蛋白质免疫印迹法（Western blot）、实时定量PCR和免疫组织化学法测定靶基因的表达水平。与年龄匹配的正常孕妇相比，35岁以上稽留流产患者胎盘绒毛中miR-98显著上调。miR-98的上调抑制了人滋养层细胞HTR-8/SVneo在体外的增殖、活性和迁移。miR-98可与生长分化因子6（GDF6）和含FAPP结构域蛋白2（FAPP2）的信使核糖核酸（mRNA）的3'-非翻译区（UTR）结合，并负向调节它们的表达。miR-98的下调促进细胞增殖，然后敲低GDF6或FAPP2可抑制miR-98介导的细胞增殖。与正常胎盘组织相比，稽留流产患者胎盘绒毛中GDF6和FAPP2的表达降低。稽留流产中miR-98的表达与GDF6和FAPP2的表达呈相反关系。miR-98的过表达与稽留流产的发生有关。miR-98部分通过靶向GDF6和FAPP2来阻止滋养层细胞的增殖、活力和迁移。
