环状 LRRK1 通过调控 PI3K/AKT 信号通路靶向 miR-223-3p 抑制滋养层细胞的增殖、迁移和侵袭。
CircLRRK1 targets miR-223-3p to inhibit the proliferation, migration and invasion of trophoblast cells by regulating the PI3K/AKT signaling pathway.
机构信息
Department of Obstetrics, Yantaishan Hospital of Yantai, Yantai, 264001, Shandong, China.
Department of Gynaecology, Qingdao Women and Children's Hospital, Qingdao, 266011, Shandong, China.
出版信息
Placenta. 2021 Jan 15;104:110-118. doi: 10.1016/j.placenta.2020.12.003. Epub 2020 Dec 7.
INTRODUCTION
Many studies have shown that circular RNAs (circRNAs) are related to the occurrence of preeclampsia (PE). However, the role of circLRRK1 in the progression of PE is unclear.
METHODS
The identification and localization of circLRRK1 were verified by Actinomycin D (ActD) assay, Ribonuclease R (RNase R) digestion assay and subcellular localization assay. Moreover, the proliferation of trophoblast cells was detected by 3-(45)-dimethylthiahiazo (-z-y1)-35-di-phenytetrazoliumromide (MTT) assay and colony formation assay. Furthermore, the migration and invasion of trophoblast cells were determined by wound healing assay and transwell assay. Meanwhile, Western blot (WB) analysis was used to examine the protein levels of migration markers and PI3K/AKT signaling pathway markers. In addition, the interaction between circLRRK1 and miR-223-3p was confirmed by dual-luciferase reporter assay and biotin-labeled RNA pull-down assay.
RESULTS
Our results showed that circLRRK1 was significantly highly expressed in PE patients. Silenced circLRRK1 could markedly enhance the proliferation, migration and invasion of trophoblast cells. Additionally, we found that circLRRK1 could target miR-223-3p. MiR-223-3p overexpression also promoted the proliferation, migration and invasion of trophoblast cells. The rescue experiments revealed that miR-223-3p inhibitor could reverse the promoting effect of circLRRK1 silencing on the proliferation, migration and invasion of trophoblast cells. Furthermore, circLRRK1 silencing could activate the PI3K/AKT signaling pathway by targeting miR-223-3p.
DISCUSSION
CircLRRK1 could suppress the proliferation, migration and invasion of trophoblast cells by regulating the PI3K/AKT signaling pathway via targeting miR-223-3p, suggesting that circLRRK1 might be a potential biomarker for the treatment of PE.
简介
许多研究表明，环状 RNA（circRNAs）与子痫前期（PE）的发生有关。然而，circLRRK1 在 PE 进展中的作用尚不清楚。
方法
通过放线菌素 D（ActD）试验、核糖核酸酶 R（RNase R）消化试验和亚细胞定位试验验证 circLRRK1 的鉴定和定位。此外，通过 3-(45)-二甲基噻唑 (-z-y1)-35-二苯四唑溴盐（MTT）试验和集落形成试验检测滋养细胞的增殖。进一步通过划痕愈合试验和 Transwell 试验测定滋养细胞的迁移和侵袭。同时，通过 Western blot（WB）分析检测迁移标志物和 PI3K/AKT 信号通路标志物的蛋白水平。此外，通过双荧光素酶报告基因试验和生物素标记的 RNA 下拉试验证实 circLRRK1 与 miR-223-3p 之间的相互作用。
结果
我们的结果表明，circLRRK1 在 PE 患者中显著高表达。沉默 circLRRK1 可显著增强滋养细胞的增殖、迁移和侵袭。此外，我们发现 circLRRK1 可以靶向 miR-223-3p。miR-223-3p 过表达也促进了滋养细胞的增殖、迁移和侵袭。挽救实验表明，miR-223-3p 抑制剂可以逆转 circLRRK1 沉默对滋养细胞增殖、迁移和侵袭的促进作用。此外，circLRRK1 沉默通过靶向 miR-223-3p 可以激活 PI3K/AKT 信号通路。
讨论
circLRRK1 通过靶向 miR-223-3p 调控 PI3K/AKT 信号通路，抑制滋养细胞的增殖、迁移和侵袭，提示 circLRRK1 可能是治疗 PE 的潜在生物标志物。
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