Center for Reproductive Medicine, Renmin Hospital of Wuhan University, Wuhan, 430060, China.
Department of Obstetrics, Renmin Hospital of Wuhan University, Wuhan, 430060, China.
Placenta. 2024 Jun;151:27-36. doi: 10.1016/j.placenta.2024.04.006. Epub 2024 Apr 18.
This study aimed to screen circRNAs involved in gestational diabetes mellitus (GDM)-related macrosomia. One differentially expressed circRNA (DEC), hsa_circ_0024838, was further tested for its potential role and mechanism in trophoblasts.
DECs in GDM were selected through GSE182737 and GSE194119. The targets were predicted for DECs and microRNAs (miRNAs), to complete the construction of the circRNA-miRNA-gene network. Functional annotation and related biological pathway enrichment analysis were performed on the target genes of miRNAs in the network. Subsequently, the expression levels of hsa_circ_0024838, miR-543, and HIF1A mRNA were identified by real-time quantitative real-time PCR (RT-qPCR) in GDM patients. Trophoblast activity was assessed via CCK-8 assay, apoptosis assay, and Matrigel invasion assay. Finally, interactions between miR-543 and either hsa_circ_0024838 or HIF1A were confirmed using dual-luciferase reporter assays.
A GDM-related circRNA-miRNA-genes interaction network was constructed, consisting of 35 circRNAs, 46 miRNAs, and 122 target genes. Functional enrichment revealed that the enriched pathways were involved in GDM. Hsa_circ_0024838 and HIF1A mRNA expression levels were upregulated in GDM, while miR-543 expression levels were downregulated. A significant positive correlation between hsa_circ_0024838 and newborn weight was observed. Both hsa_circ_0024838 and HIF1A possessed binding sites for miR-543. Overexpressing hsa_circ_0024838 in high-glucose (HG)-cultured trophoblasts can partially reverse HG-induced reduction in trophoblast cell proliferation/migration and increase apoptosis. But this reversal can be negated by co-transfection with miR-543 mimics. The effects of miR-543 can be counteracted by HIF1A.
Hsa_circ_0024838 can regulate the expression of HIF1A by interacting with miR-543. This regulates the HIF1A signaling pathway and enhance vitality in trophoblast cells.
本研究旨在筛选与妊娠糖尿病（GDM）相关巨大儿相关的 circRNA。进一步检测了一个差异表达的 circRNA（DEC），hsa_circ_0024838，以研究其在滋养细胞中的潜在作用和机制。
通过 GSE182737 和 GSE194119 筛选 GDM 中的 DEC。针对 DEC 和 microRNAs（miRNAs）进行靶标预测，以完成 circRNA-miRNA-基因网络的构建。对网络中 miRNA 的靶基因进行功能注释和相关生物途径富集分析。随后，通过实时定量聚合酶链反应（RT-qPCR）检测 GDM 患者中 hsa_circ_0024838、miR-543 和 HIF1A mRNA 的表达水平。通过 CCK-8 测定、凋亡测定和 Matrigel 侵袭测定评估滋养细胞活性。最后，通过双荧光素酶报告基因测定证实 miR-543 与 hsa_circ_0024838 或 HIF1A 之间的相互作用。
构建了一个与 GDM 相关的 circRNA-miRNA-基因相互作用网络，该网络由 35 个 circRNAs、46 个 miRNAs 和 122 个靶基因组成。功能富集显示，富集途径与 GDM 有关。GDM 中 hsa_circ_0024838 和 HIF1A mRNA 的表达水平上调，而 miR-543 的表达水平下调。hsa_circ_0024838 与新生儿体重呈显著正相关。hsa_circ_0024838 和 HIF1A 均具有 miR-543 的结合位点。在高葡萄糖（HG）培养的滋养细胞中过表达 hsa_circ_0024838 可部分逆转 HG 诱导的滋养细胞增殖/迁移减少和凋亡增加。但这一逆转可以通过共转染 miR-543 模拟物来否定。miR-543 的作用可以被 HIF1A 抵消。
hsa_circ_0024838 可以通过与 miR-543 相互作用来调节 HIF1A 的表达。这调节了 HIF1A 信号通路并增强了滋养细胞的活力。
