Hsa-miR-3928–3p targets the CCL3/CCR5 axis to induce amniotic epithelial cell senescence involved in labor initiation

IF 3 2区 医学 Q2 DEVELOPMENTAL BIOLOGY
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Abstract

Introduction

Senescence in human amniotic epithelial cells (hAECs) and increased sterile inflammation in the amniotic cavity can lead to the initiation of term labor (TL). We investigated the possible roles of hsa-miR-3928–3p and chemokine ligand 3 (CCL3) in labor initiation and the underlying molecular mechanisms.

Methods

Microarray chip screening was used to analyse the differential expression of miRNAs in amniotic fluid exosomes from women in TL and term not-in-labor. The GEO and miRWalk databases were used to identify differential genes, and a dual luciferase assay was used to verify the relationship. Reverse transcription quantitative PCR (RT-qPCR) and immunofluorescence were used to determine the expression and localization of CCL3/CCR5 in fetal membranes. RT-qPCR and western blotting were used to detect the expression of CCL3/CCR5 in hAECs with hsa-miR-3928–3p knockdown/overexpression. Cell counting kit 8, flow cytometry, EdU proliferation, senescence-associated β-galactosidase, and enzyme-linked immunosorbent assays were performed to detect the impact of hsa-miR-3928–3p on hAEC function.

Results

hsa-miR-3928–3p expression was downregulated in TL. CCL3 (macrophage inflammatory protein-1α) was identified as a differentially expressed target gene. hsa-miR-3928–3p targeted the 3′ UTR of CCL3. Downregulation of hsa-miR-3928–3p expression increased CCL3 expression. CCL3, via its CCR5 receptor, decreased the proliferation, but increased the senescence, apoptosis rate, secretion of inflammatory factors (IL-8, TNF-α, and IL-6), and expression of senescence-associated protein p21 in hAECs.

Discussion

hsa-miR-3928–3p negatively regulates CCL3, promoting hAEC senescence through the CCL3-CCR5 axis and inducing signals for labor initiation. These findings provide novel insights for labor initiation in clinical settings.

Abstract Image

Hsa-miR-3928-3p 以 CCL3/CCR5 轴为靶标,诱导羊膜上皮细胞衰老参与分娩启动
引言人类羊膜上皮细胞(hAECs)的衰老和羊膜腔无菌炎症的增加可导致临产(TL)的开始。我们研究了 hsa-miR-3928-3p 和趋化因子配体 3 (CCL3) 在临产中的可能作用及其潜在的分子机制。利用 GEO 和 miRWalk 数据库确定差异基因,并利用双荧光素酶试验验证两者之间的关系。反转录定量 PCR(RT-qPCR)和免疫荧光用于确定 CCL3/CCR5 在胎膜中的表达和定位。采用 RT-qPCR 和 Western 印迹法检测 hsa-miR-3928-3p 基因敲除/缺失表达的 hAECs 中 CCL3/CCR5 的表达。为了检测 hsa-miR-3928-3p 对 hAEC 功能的影响,还进行了细胞计数试剂盒 8、流式细胞术、EdU 增殖、衰老相关β-半乳糖苷酶和酶联免疫吸附试验。CCL3(巨噬细胞炎症蛋白-1α)被确定为差异表达的靶基因。下调 hsa-miR-3928-3p 的表达可增加 CCL3 的表达。讨论hsa-miR-3928-3p负调控CCL3,通过CCL3-CCR5轴促进hAEC衰老并诱导分娩启动信号。这些发现为临床中的分娩启动提供了新的见解。
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来源期刊
Placenta
Placenta 医学-发育生物学
CiteScore
6.30
自引率
10.50%
发文量
391
审稿时长
78 days
期刊介绍: Placenta publishes high-quality original articles and invited topical reviews on all aspects of human and animal placentation, and the interactions between the mother, the placenta and fetal development. Topics covered include evolution, development, genetics and epigenetics, stem cells, metabolism, transport, immunology, pathology, pharmacology, cell and molecular biology, and developmental programming. The Editors welcome studies on implantation and the endometrium, comparative placentation, the uterine and umbilical circulations, the relationship between fetal and placental development, clinical aspects of altered placental development or function, the placental membranes, the influence of paternal factors on placental development or function, and the assessment of biomarkers of placental disorders.
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