Department of Obstetrics and Gynecology, The First Hospital of Jilin University, Jilin University, 71 Xinmin Dajie, Chaoyang District, Changchun, 130021, Jilin, China.
Department of Sports Medicine, The First Hospital of Jilin University, Jilin University, Changchun, China.
Reprod Sci. 2022 Oct;29(10):2829-2841. doi: 10.1007/s43032-021-00804-y. Epub 2021 Nov 29.
Accumulating evidence shows that impaired spiral artery remodeling, placental dysfunction, and insufficient trophoblast infiltration contribute to the etiology and pathogenesis of pre-eclampsia (PE). circRNAs are a class of endogenous non-coding RNAs implicated in the pathogenesis of many diseases, including PE. This study aims to investigate the role of circRNA hsa_circ_0008726 in regulating the migration and invasion of extravillous trophoblast cells. RNase R assay was performed to confirm that circ_0008726 was a circular transcript. The expression of circ_0008726, RYBP, and miR-345-3p was examined by qRT-PCR. The functional interaction between miR-345-3p and circ_0008726 or RYBP was confirmed using dual-luciferase reporter assay and RNA immunoprecipitation (RIP). Cell migration and invasion ability was analyzed by Transwell assays. Western blot was used for the quantification of RYBP protein level. Circ_0008726 expression was significantly increased in PE placenta tissues as compared with normal placenta tissues. Circ_0008726 was resistant to RNase R digestion and was predominately located in the cytoplasm of HTR-8/SVneo cells. Silencing circ_0008726 promoted cell migration and EMT (epithelial-mesenchymal transition), while circ_0008726 overexpression suppressed these processes. Mechanistically, circ_0008726 sponged miR-345-3p to negatively regulate its expression, and miR-345-3p negatively modulated the expression of RYBP. In PE samples, the expression level of circ_0008726 was negatively correlated with miR-345-3p level, but was positively correlated with RYBP expression. Transfection of miR-345-3p mimic or RYBP knockdown counteracted the effects of circ_0008726 overexpression on cell migration and EMT. Our data demonstrate the upregulation of circ_0008726 in PE placenta, which inhibits the migration, invasion, and EMT of HTR-8/SVneo cells by targeting miR-345-3p/RYBP axis. These data suggest that circ_0008726 could be a potential biomarker and therapeutic target for PE.
越来越多的证据表明，螺旋动脉重塑受损、胎盘功能障碍和滋养细胞浸润不足导致子痫前期 (PE) 的病因和发病机制。circRNAs 是一类内源性非编码 RNA，与许多疾病的发病机制有关，包括 PE。本研究旨在探讨 circRNA hsa_circ_0008726 在调节绒毛外滋养细胞迁移和侵袭中的作用。通过 RNase R 分析实验确证 circ_0008726 是一种环状转录本。通过 qRT-PCR 检测 circ_0008726、RYBP 和 miR-345-3p 的表达。通过双荧光素酶报告基因检测和 RNA 免疫沉淀 (RIP) 实验验证 miR-345-3p 与 circ_0008726 或 RYBP 的功能相互作用。通过 Transwell 实验分析细胞迁移和侵袭能力。通过 Western blot 检测 RYBP 蛋白水平的定量。与正常胎盘组织相比，PE 胎盘组织中 circ_0008726 的表达明显增加。Circ_0008726 对 RNase R 消化有抗性，并且主要位于 HTR-8/SVneo 细胞的细胞质中。沉默 circ_0008726 促进细胞迁移和 EMT（上皮-间充质转化），而过表达 circ_0008726 则抑制这些过程。机制上，circ_0008726 作为 miR-345-3p 的海绵体，负调控其表达，而 miR-345-3p 负调控 RYBP 的表达。在 PE 样本中，circ_0008726 的表达水平与 miR-345-3p 水平呈负相关，但与 RYBP 表达呈正相关。转染 miR-345-3p 模拟物或 RYBP 敲低可逆转 circ_0008726 过表达对细胞迁移和 EMT 的影响。我们的数据表明，PE 胎盘组织中 circ_0008726 的上调通过靶向 miR-345-3p/RYBP 轴抑制 HTR-8/SVneo 细胞的迁移、侵袭和 EMT。这些数据表明，circ_0008726 可能是 PE 的潜在生物标志物和治疗靶点。
