Department of Obstetrics, The Affiliated Huaian No. 1 People's Hospital of Nanjing Medical University, No. 1 Huanghe West Road, Huai'an, 223300, Jiangsu, China.
Department of Pathology, The Affiliated Huaian No. 1 People's Hospital of Nanjing Medical University, Huai, 223300, China.
Reprod Sci. 2022 Jan;29(1):100-109. doi: 10.1007/s43032-021-00713-0. Epub 2021 Aug 30.
Noncoding circular RNAs (circRNAs) have participated in the progression of preeclampsia (PE) via inhibiting microRNAs (miRNAs) to regulate gene expression. This study was designed to explore the miRNA/mRNA mechanism of hsa_circ_0007121 (circ_0007121) in PE. The expression detection of circ_0007121, microRNA-421 (miR-421), and zinc finger E-box binding homeobox 1 (ZEB1) was performed by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was analyzed by Cell Counting Kit-8 (CCK-8) assay. Transwell assay was used to determine cell migration and invasion. Cell apoptosis was evaluated using flow cytometry. The protein levels of epithelial-mesenchymal transition (EMT) markers and ZEB1 were measured via western blot. The interaction between miR-421 and circ_0007121 or ZEB1 was validated by dual-luciferase reporter assay. The expression detection indicated that circ_0007121 was downregulated in PE patients and the clinical data revealed that circ_0007121 was related to PE. The upregulation of circ_0007121 promoted cell proliferation, migration, invasion, and EMT in trophoblastic cells. Furthermore, circ_0007121 was identified as a sponge of miR-421 and the function of circ_0007121 was dependent on the sponge effect on miR-421. Moreover, ZEB1 was a target of miR-421 and circ_0007121/miR-421 axis could regulate the expression of ZEB1. In addition, miR-421 overexpression repressed trophoblastic cell behaviors through downregulating the ZEB1 level. Altogether, circ_0007121 contributed to the development of trophoblastic cells by regulating the miR-421/ZEB1 axis.
非编码环状 RNA（circRNA）通过抑制 microRNA（miRNA）来调节基因表达，从而参与子痫前期（PE）的进展。本研究旨在探讨 hsa_circ_0007121（circ_0007121）在 PE 中的 miRNA/mRNA 机制。通过实时定量聚合酶链反应（qRT-PCR）检测 circ_0007121、microRNA-421（miR-421）和锌指 E 盒结合同源盒 1（ZEB1）的表达。通过细胞计数试剂盒-8（CCK-8）测定细胞增殖。使用 Transwell 测定法确定细胞迁移和侵袭。通过流式细胞术评估细胞凋亡。通过 Western blot 测定上皮-间充质转化（EMT）标志物和 ZEB1 的蛋白水平。通过双荧光素酶报告基因测定验证 miR-421 与 circ_0007121 或 ZEB1 的相互作用。表达检测表明 circ_0007121 在 PE 患者中下调，临床数据表明 circ_0007121 与 PE 相关。circ_0007121 的上调促进滋养细胞的增殖、迁移、侵袭和 EMT。此外，circ_0007121 被鉴定为 miR-421 的海绵，circ_0007121 的功能依赖于对 miR-421 的海绵作用。此外，ZEB1 是 miR-421 的靶标，circ_0007121/miR-421 轴可以调节 ZEB1 的表达。此外，miR-421 的过表达通过下调 ZEB1 水平抑制滋养细胞行为。总之，circ_0007121 通过调节 miR-421/ZEB1 轴促进滋养细胞的发展。
