Department of Obstetrics, Jingmen First People's Hospital, Jingmen City, 448000, Hubei Province, China.
Department of Gynecology, Jingmen First People's Hospital, Jingmen City, 448000, Hubei Province, China.
Reprod Biol. 2021 Dec;21(4):100571. doi: 10.1016/j.repbio.2021.100571. Epub 2021 Nov 3.
Preeclampsia (PE) is the leading cause of maternal deaths in primipara. It is mainly characterized by defect migration and invasion of trophoblast cells. Circular RNAs (circRNAs) have been widely reported to be associated with PE progression. This study is designed to explore the role and mechanism of circ_0032962 on trophoblast cell behavior. Circ_0032962, microRNA-326 (miR-326), and Pre-B-cell leukemia homeobox 3 (PBX3) levels were measured by real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferation ability, migration, and invasion were measured by Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), Colony formation, wound healing, and transwell assays. Protein levels of E-cadherin, Vimentin, N-cadherin, and PBX3 were examined by western blot assay. The binding relationship between miR-326 and circ_0032962 or PBX3 was predicted by circular RNA Interactome or Starbase and then verified by a dual-luciferase reporter assay. Circ_0032962 and PBX3 levels were declined in placenta tissues from preeclampsia patients, and miR-326 was elevated. Apart from that, circ_0032962 knockdown could suppress cell proliferation ability, migration, invasion, and epithelial-mesenchymal transition (EMT) in trophoblast cells. Mechanically, circ_0032962 could affect PBX3 expression through sponging miR-326. Circ_0032962 could contribute to trophoblast cell growth ability and metastasis partly by regulating the miR-326/PBX3 axis, providing a novel insight into the pathogenesis and treatment of PE.
子痫前期 (PE) 是初产妇死亡的主要原因。它主要表现为滋养细胞迁移和侵袭缺陷。环状 RNA (circRNA) 已被广泛报道与 PE 进展有关。本研究旨在探讨 circ_0032962 对滋养细胞行为的作用和机制。通过实时定量聚合酶链反应 (RT-qPCR) 测量 circ_0032962、microRNA-326 (miR-326) 和 Pre-B 细胞白血病 homeobox 3 (PBX3) 的水平。通过细胞计数试剂盒-8 (CCK-8)、5-乙炔基-2'-脱氧尿苷 (EdU)、集落形成、划痕愈合和 Transwell 测定测量细胞增殖能力、迁移和侵袭。通过 Western blot 测定检测 E-钙粘蛋白、波形蛋白、N-钙粘蛋白和 PBX3 的蛋白水平。通过环状 RNA 相互作用或 Starbase 预测 miR-326 与 circ_0032962 或 PBX3 的结合关系,然后通过双荧光素酶报告基因检测验证。子痫前期患者胎盘组织中 circ_0032962 和 PBX3 水平下降,miR-326 水平升高。除此之外,circ_0032962 敲低可抑制滋养细胞的增殖能力、迁移、侵袭和上皮-间充质转化 (EMT)。从机制上讲,circ_0032962 可以通过海绵 miR-326 来影响 PBX3 的表达。circ_0032962 可以通过调节 miR-326/PBX3 轴部分促进滋养细胞生长能力和转移,为 PE 的发病机制和治疗提供新的见解。