Department of Obstetrics, Qingpu Branch of Zhongshan Hospital Affiliated to Fudan University, No.1158 Park East Road, Qingpu District, Shanghai, 201700, China.
Reprod Sci. 2022 Mar;29(3):923-935. doi: 10.1007/s43032-021-00715-y. Epub 2021 Aug 30.
Preeclampsia is the main reason for maternal and fetal deaths during the second half of pregnancy. Trophoblast cells play a pivotal role in preeclampsia progression. Circular RNA (circRNA) circ_0111277 has been reported to be related to the development of trophoblast cells. This study is designed to explore the role and mechanism of circ_0111277 on trophoblast cell behavior in preeclampsia. Circ_0111277, microRNA-424-5p (miR-424-5p), and nuclear factor of activated T-cell 5 (NFAT5) levels were measured by real-time quantitative polymerase chain reaction (RT-qPCR). Cell viability, migration, invasion, and angiogenesis were measured by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay, transwell assay, tube formation assay, and wound healing assay. Protein levels of matrix metallopeptidase 2 (MMP2), vascular endothelial growth factor-A (VEGF-A), NFAT5, phospho-phosphatidylinositol 3 kinase (p-PI3K), PI3K, phospho-protein kinase B (p-AKT), and AKT were examined by western blot assay. The binding relationship between miR-424-5p and circ_0111277 or NFAT5 was predicted by circBank or starBase and then verified by a dual-luciferase reporter assay. Circ_0111277 and NFAT5 expression were increased in placenta tissues of preeclampsia patients, and miR-424-5p was decreased. Moreover, circ_0111277 knockdown could boost cell viability, migration, invasion, and angiogenesis in trophoblast cells. The mechanical analysis discovered that circ_0111277 acted as a sponge of miR-424-5p to regulate NFAT5 expression. Besides, circ_0111277 silencing promoted the PI3K/AKT signaling pathway in trophoblast cells. Circ_0111277 downregulation could facilitate cell growth and metastasis in trophoblast cells partly by regulating the miR-424-5p/NFAT5 axis, providing an underlying circRNA-targeted therapy for preeclampsia.
子痫前期是妊娠后半期母婴死亡的主要原因。滋养细胞在子痫前期的进展中起着关键作用。环状 RNA（circRNA）circ_0111277 已被报道与滋养细胞的发育有关。本研究旨在探讨 circ_0111277 对滋养细胞行为在子痫前期中的作用和机制。通过实时定量聚合酶链反应（RT-qPCR）测量 circ_0111277、微小 RNA-424-5p（miR-424-5p）和活化 T 细胞核因子 5（NFAT5）的水平。通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）测定细胞活力、迁移、侵袭和血管生成，通过 Transwell 测定、管形成测定和划痕愈合测定。通过 Western blot 测定基质金属蛋白酶 2（MMP2）、血管内皮生长因子-A（VEGF-A）、NFAT5、磷酸化磷脂酰肌醇 3 激酶（p-PI3K）、PI3K、磷酸化蛋白激酶 B（p-AKT）和 AKT 的蛋白水平。通过 circBank 或 starBase 预测 miR-424-5p 与 circ_0111277 或 NFAT5 的结合关系，然后通过双荧光素酶报告基因测定验证。子痫前期患者胎盘组织中 circ_0111277 和 NFAT5 的表达增加，而 miR-424-5p 的表达降低。此外，circ_0111277 敲低可促进滋养细胞的细胞活力、迁移、侵袭和血管生成。力学分析发现，circ_0111277 作为 miR-424-5p 的海绵调节 NFAT5 的表达。此外，circ_0111277 沉默促进了滋养细胞中的 PI3K/AKT 信号通路。circ_0111277 下调可能部分通过调节 miR-424-5p/NFAT5 轴促进滋养细胞的细胞生长和转移，为子痫前期提供了一种潜在的基于环状 RNA 的治疗方法。
