Department of Obstetrics and Gynecology, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, 250021, China; Department of Obstetrics and Gynecology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, 250021, China; Department of Obstetrics and Gynecology, Feixian County People's Hospital, Linyi, Shandong, 273400, China.
Department of Obstetrics and Gynecology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, 250021, China.
Placenta. 2024 Oct;156:98-107. doi: 10.1016/j.placenta.2024.09.008. Epub 2024 Sep 10.
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.
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.
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.
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.
人羊膜上皮细胞（hAECs）衰老和羊膜腔内无菌性炎症增加可导致足月产（TL）的启动。我们研究了 hsa-miR-3928-3p 和趋化因子配体 3（CCL3）在分娩启动中的可能作用及其潜在的分子机制。
采用微阵列芯片筛选技术分析 TL 和足月未临产孕妇羊水中外泌体中 miRNA 的差异表达。利用 GEO 和 miRWalk 数据库鉴定差异基因，并通过双荧光素酶报告基因实验验证。逆转录定量 PCR（RT-qPCR）和免疫荧光法检测胎儿膜中 CCL3/CCR5 的表达和定位。采用 RT-qPCR 和 Western blot 检测 hAECs 中 hsa-miR-3928-3p 敲低/过表达后 CCL3/CCR5 的表达。采用细胞计数试剂盒 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检测 hsa-miR-3928-3p 对 hAEC 功能的影响。
TL 中 hsa-miR-3928-3p 的表达下调。鉴定出 CCL3（巨噬细胞炎性蛋白-1α）为差异表达的靶基因。hsa-miR-3928-3p 靶向 CCL3 的 3'UTR。下调 hsa-miR-3928-3p 的表达增加了 CCL3 的表达。CCL3 通过其 CCR5 受体降低 hAEC 的增殖能力，但增加衰老、凋亡率、炎症因子（IL-8、TNF-α和 IL-6）的分泌以及衰老相关蛋白 p21 的表达。
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。这些发现为临床分娩启动提供了新的见解。
