Zhou Meixi, Yu Xiang, Li Chenping, Lou Lejing, Yang Song, Cai Jihao, Cai Chang
Department of Gynecology and Obstetrics, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
Department of pulmonary and critical care medicine, Yongjia Hospital of traditional Chinese Medicine, Wenzhou, China.
Am J Reprod Immunol. 2023 Mar;89(3):e13657. doi: 10.1111/aji.13657. Epub 2022 Nov 29.
Preeclampsia (PE) is the main factor threatening the life of primipara. Defective migration and invasion of trophoblast cells was one of the causes of PE. Circ_0111277 had been reported to be related to the development of PE, but the mechanism of its effect on trophoblast cells needed further study.
The expression of circ_0111277, microRNA-188-3p (miR-188-3p) and grainyhead-like 2 (GRHL2) mRNA were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell Counting Kit-8 (CCK-8) assay, 5-Ethynyl-20-deoxyuridine (EdU) and colony formation assay were used to examine cell proliferation ability. Tube formation and transwell assay were performed to assess the angiogenesis and metastasis ability of cells. Western blot was applied to measure the levels of epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin and Vimentin) and GRHL2 protein. The relationship between miR-188-3p and circ_0111277 or GRHL2 was verified by the dual luciferase reporter experiment.
Circ_0111277 and GRHL2 were elevated, and miR-188-3p was declined in PE patients. Overexpression of circ_0111277 could inhibit the proliferation, angiogenesis, migration, invasion and EMT of trophoblast cells (HTR-8/Svneo). Circ_0111277 was the molecular sponge of miR-188-3p. MiR-188-3p up-regulation could reduce the inhibition of HTR-8/Svneo cell growth caused by overexpression of circ_0111277. GRHL2 was a target gene of miR-188-3p, and GRHL2 silencing relieved the adverse effects of miR-188-3p inhibitors on HTR-8/Svneo. In general, circ_0111277 up-regulated GRHL2 expression through sponge miR-188-3p.
Highly expressed circ_0111277 up-regulated the expression of GRHL2 through sponge miR-188-3p, thereby inhibiting trophoblast cells function, which suggested a new molecular mechanism for the pathogenesis of PE.
子痫前期（PE）是威胁初产妇生命的主要因素。滋养层细胞迁移和侵袭缺陷是PE的病因之一。已有报道称Circ_0111277与PE的发生发展有关，但其对滋养层细胞的作用机制尚需进一步研究。
采用定量实时聚合酶链反应（qRT-PCR）检测Circ_0111277、微小RNA-188-3p（miR-188-3p）和颗粒头样蛋白2（GRHL2）mRNA的表达。使用细胞计数试剂盒-8（CCK-8）检测法、5-乙炔基-2'-脱氧尿苷（EdU）和集落形成试验检测细胞增殖能力。进行管腔形成试验和Transwell试验评估细胞的血管生成和转移能力。采用蛋白质免疫印迹法检测上皮-间质转化（EMT）相关蛋白（E-钙黏蛋白和波形蛋白）和GRHL2蛋白的水平。通过双荧光素酶报告基因实验验证miR-188-3p与Circ_0111277或GRHL2之间的关系。
PE患者中Circ_0111277和GRHL2升高，miR-188-3p降低。Circ_0111277过表达可抑制滋养层细胞（HTR-8/Svneo）的增殖、血管生成、迁移、侵袭和EMT。Circ_0111277是miR-188-3p的分子海绵。上调miR-188-3p可减轻Circ_0111277过表达对HTR-8/Svneo细胞生长的抑制作用。GRHL2是miR-188-3p的靶基因，GRHL2沉默可缓解miR-该研究旨在探讨C1rc_0111277在子痫前期发病机制中的作用及其分子机制。
高表达的Circ_0111277通过海绵化miR-188-3p上调GRHL2表达，从而抑制滋养层细胞功能，这提示了PE发病机制的一种新的分子机制。
