Tan Wang, Fu Huijia, Zhou Xiaobo, Duan Yuhan, Yin Nanlin, Huang Jiayu, Liu Xiru
Reproductive Medicine Center, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
The Chongqing Key Laboratory of Translational Medicine in Major Metabolic Diseases, Chongqing, China.
J Gene Med. 2022 May;24(5):e3416. doi: 10.1002/jgm.3416. Epub 2022 Mar 5.
Inadequate trophoblast invasion is associated with preeclampsia (PE). Ankyrin repeat domain protein 37 (ANKRD37) has been reported to be abnormally expressed in PE placentas. However, the role of ANKRD37 in trophoblasts has not been investigated. We aimed to determine the functions of ANKRD37 in PE and to explore the molecular mechanisms.
Here, fluorescence in situ hybridization, immunohistochemistry, Western blotting and quantitative real-time polymerase chain reaction were used to detect protein and mRNA expression levels. Cell counting kit-8 assay, 5-ethynyl-2'-deoxyuridine assay, flow cytometry, wound healing assay, transwell assay and RNA sequencing were performed to investigate the role of ANKRD37 and the underlying mechanism in HTR8/SVneo and JEG-3 cells, and extravillous explant cultures were used to evaluate the migration and invasion abilities of extravillous cytotrophoblasts.
We found that ANKRD37 expression was upregulated in PE placentas compared to normal pregnancy placentas. ANKRD37 knockdown enhanced trophoblast migration and invasion, promoted extravillous explant outgrowth, and regulated the expression of key invasion proteins, whereas ANKRD37 overexpression exerted the opposite effects. RNA sequencing indicated that nuclear factor-kappa B (NF-κB) was the potential downstream pathway of ANKRD37, which was confirmed by the change in p-p65 and p-IκBα expression in JEG-3 and HTR8/SVneo cells.
Our findings suggest that high expression of ANKRD37 inhibits trophoblast cell migration and invasion possibly via the NF-κB pathway, and may be related to the development of PE.
滋养层细胞浸润不足与子痫前期（PE）相关。据报道，锚蛋白重复结构域蛋白37（ANKRD37）在PE胎盘组织中表达异常。然而，ANKRD37在滋养层细胞中的作用尚未得到研究。我们旨在确定ANKRD37在PE中的功能并探索其分子机制。
在这里，采用荧光原位杂交、免疫组织化学、蛋白质印迹法和定量实时聚合酶链反应来检测蛋白质和mRNA表达水平。进行细胞计数试剂盒-8检测、5-乙炔基-2'-脱氧尿苷检测、流式细胞术、伤口愈合检测、Transwell检测和RNA测序，以研究ANKRD37在HTR8/SVneo和JEG-3细胞中的作用及其潜在机制，并且使用绒毛外植体培养来评估绒毛外细胞滋养层细胞的迁移和侵袭能力。
我们发现，与正常妊娠胎盘相比，ANKRD37在PE胎盘组织中的表达上调。ANKRD37基因敲低增强了滋养层细胞的迁移和侵袭能力，促进了绒毛外植体的生长，并调节了关键侵袭蛋白的表达，而ANKRD37过表达则产生相反的效果。RNA测序表明，核因子-κB（NF-κB）是ANKRD37潜在的下游信号通路，这在JEG-3和HTR8/SVneo细胞中p-p65和p-IκBα表达的变化中得到证实。
我们的研究结果表明，ANKRD37的高表达可能通过NF-κB信号通路抑制滋养层细胞的迁移和侵袭，并且可能与PE的发生发展有关。
