Background Preeclampsia (PE) is a prevalent pregnancy disorder that has been one of the leading causes of maternal and perinatal mortality worldwide. Circular RNAs (circRNAs) have recently considered as important regulators in PE pathogenesis. In the current study, we aimed to explore the impact and mechanisms of circRNA zinc finger DHHC-type palmitoyltransferase 20 (circZDHHC20) in PE pathogenesis. Methods RNase R assay and reverse transcription with Oligo(dT)18 primers were performed to confirm that circZDHHC20 was indeed circular transcript. The expression of circZDHHC20, grainyhead-like 2 (GRHL2) and miR-144 were assessed by quantitative real-time polymerase chain reaction (qRT-PCR). Subcellular localization assay was used to determine whether circZDHHC20 was predominantly present in the cytoplasm. The target correlations between miR-144 and circZDHHC20 or GRHL2 were confirmed using dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Cell proliferation, migration, and invasion were detected by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetr-azolium (MTS), wound healing and transwell assays, respectively. Western blot was used for the quantification of GRHL2 protein level. Results Our data indicated that circZDHHC20 was up-regulated and miR-144 was down-regulated in PE placenta. CircZDHHC20 sequestered miR-144 by acting as a miR-144 sponge. CircZDHHC20 overexpression repressed trophoblast cell proliferation, migration, and invasion, while its knockdown exerted opposite effects. Moreover, miR-144 mediated the regulation of circZDHHC20 on trophoblast cell behaviors. GRHL2 was directly targeted and inhibited by miR-144. MiR-144 exerted regulatory effects on trophoblast cell proliferation, migration and invasion by GRHL2. Furthermore, circZDHHC20 modulated GRHL2 expression through sponging miR-144. Conclusion Our study suggested that a high level of circZDHHC20 inhibited the proliferation, migration, and invasion in trophoblast cells at least partially through sponging miR-144 and up-regulating GRHL2, providing a novel mechanism of PE pathogenesis.

中文翻译：

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CircZDHHC20 通过 miR-144/GRHL2 轴抑制滋养层细胞的增殖、迁移和侵袭。

背景 先兆子痫 (PE) 是一种普遍的妊娠疾病，已成为全世界孕产妇和围产期死亡的主要原因之一。环状 RNA (circRNA) 最近被认为是 PE 发病机制中的重要调节因子。在本研究中，我们旨在探讨circRNA锌指DHHC型棕榈酰转移酶20（circZDHHC20）在PE发病机制中的影响和机制。方法 用Oligo(dT)18引物进行RNase R检测和逆转录，证实circZDHHC20确实是环状转录物。通过定量实时聚合酶链反应 (qRT-PCR) 评估 circZDHHC20、grainyhead-like 2 (GRHL2) 和 miR-144 的表达。亚细胞定位分析用于确定 circZDHHC20 是否主要存在于细胞质中。使用双荧光素酶报告基因和 RNA 免疫沉淀 (RIP) 测定证实了 miR-144 和 circZDHHC20 或 GRHL2 之间的靶标相关性。3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetr-azolium (MTS) 检测细胞增殖、迁移和侵袭。愈合和transwell检测，分别。Western印迹用于定量GRHL2蛋白水平。结果我们的数据表明，在 PE 胎盘中 circZDHHC20 上调，miR-144 下调。CircZDHHC20 通过充当 miR-144 海绵隔离 miR-144。CircZDHHC20 过表达抑制滋养层细胞的增殖、迁移和侵袭，而其敲低则产生相反的效果。此外，miR-144 介导了 circZDHHC20 对滋养层细胞行为的调节。GRHL2 被 miR-144 直接靶向和抑制。MiR-144 通过 GRHL2 对滋养层细胞的增殖、迁移和侵袭发挥调节作用。此外，circZDHHC20 通过海绵化 miR-144 调节 GRHL2 的表达。结论 我们的研究表明，高水平的 circZDHHC20 至少部分通过海绵化 miR-144 和上调 GRHL2 抑制滋养层细胞的增殖、迁移和侵袭，为 PE 发病机制提供了新的机制。