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微小RNA-214-5p通过靶向锯齿状蛋白1抑制Notch信号通路,从而抑制子痫前期中滋养层细胞的增殖、迁移和侵袭。

miR-214-5p suppresses the proliferation, migration and invasion of trophoblast cells in pre-eclampsia by targeting jagged 1 to inhibit notch signaling pathway.

作者信息

Gong Fengyan, Chai Wei, Wang Junwei, Cheng Huiyan, Shi Yuee, Cui Lifeng, Jia Guifeng

机构信息

Department of Obstetrics and Gynecology, The First Hospital of Jilin University, Changchun 130021, People's Republic of China.

Department of Obstetrics and Gynecology, The First Hospital of Jilin University, Changchun 130021, People's Republic of China.

出版信息

Acta Histochem. 2020 Apr;122(3):151527. doi: 10.1016/j.acthis.2020.151527. Epub 2020 Feb 26.

Abstract

MicroRNA-214-5p has been reported to be expressed in placental tissue and suppressed the proliferation and invasion of various tumor cells. However, the role of miR-214-5p in pre-eclampsia has not been reported. We aimed to explore the effects of miR-214-5p in proliferation, migration, and invasion of placental trophoblast cells. RT-qPCR was used to quantify the miR-214-5p expression level in placental samples and four types of trophoblast cell lines. Cell proliferation was monitored by CCK-8 and Edu staining assays. Flow cytometry was used to determine the cell cycle. Wound healing and transwell assays were performed to measure the migratory and invasive capacities in JEG-3 and BEWO cells. In addition, we investigated whether miR-214-5p targeted Jagged 1 to regulate the Notch signaling pathway to affect trophoblast cells by luciferase assay and western blot. The expression of miR-214-5p was significantly increased in the placenta of patients with PE. Moreover, the proliferation, migration, and invasion of JEG-3 cells transfected with miR-214-5p mimic were inhibited. The results were reversed when BEWO cells were transfected with miR-214-5p inhibitor. The dual-luciferase assay demonstrated that miR-214-5p directly regulated Jagged 1. The expression of the proteins associated with the Notch signaling pathway, Jagged 1, Notch 1, HEY 1 and HES 1 were all decreased when Jagged 1 was negatively regulated by miR-214-5p. miR-214-5p directly down-regulated Jagged 1 expression, then suppressed proliferation, migration, and invasion of human placental trophoblast cells by inhibiting the Notch signaling pathway.

摘要

据报道,MicroRNA-214-5p在胎盘组织中表达,并抑制多种肿瘤细胞的增殖和侵袭。然而,miR-214-5p在子痫前期中的作用尚未见报道。我们旨在探讨miR-214-5p对胎盘滋养层细胞增殖、迁移和侵袭的影响。采用RT-qPCR定量胎盘样本和四种滋养层细胞系中miR-214-5p的表达水平。通过CCK-8和Edu染色试验监测细胞增殖。流式细胞术用于测定细胞周期。进行伤口愈合试验和Transwell试验以测量JEG-3和BEWO细胞的迁移和侵袭能力。此外,我们通过荧光素酶试验和蛋白质印迹法研究miR-214-5p是否靶向Jagged 1以调节Notch信号通路来影响滋养层细胞。子痫前期患者胎盘中miR-214-5p的表达显著增加。此外,转染miR-214-5p模拟物的JEG-3细胞的增殖、迁移和侵袭受到抑制。当用miR-214-5p抑制剂转染BEWO细胞时,结果相反。双荧光素酶试验表明miR-214-5p直接调节Jagged 1。当Jagged 1受到miR-214-5p负调控时,与Notch信号通路相关的蛋白质Jagged 1、Notch 1、HEY 1和HES 1的表达均降低。miR-214-5p直接下调Jagged 1表达,然后通过抑制Notch信号通路抑制人胎盘滋养层细胞的增殖、迁移和侵袭。

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