Several array-based microRNA (miRNA) expression studies independently showed increased expression of miRNAs hsa-miR-130a-3p, -142-3p, -144-3p, -144-5p, -223-3p, -17-5p, and -30e-5p in gingiva affected by periodontal inflammation. We aimed to determine direct target genes and signaling pathways regulated by these miRNAs to identify processes relevant to gingival inflammatory responses and tissue homeostasis. We transfected miRNA mimics (mirVana) for each of the 7 miRNAs separately into human primary gingival fibroblasts cultured from 3 different donors. Following RNA sequencing, differential gene expression and second-generation gene set enrichment analyses were performed. miRNA inhibition and upregulation was validated at the transcript and protein levels using quantitative reverse transcriptase polymerase chain reaction, Western blotting, and reporter gene assays. All 7 miRNAs significantly increased expression of the gene MET proto-oncogene, receptor tyrosine kinase (MET). Expression of known periodontitis risk genes CPEB1, ABCA1, and ATP6V1C1 was significantly repressed by hsa-miR-130a-3p, -144-3p, and -144-5p, respectively. The genes WASL, ENPP5, ARL6IP1, and IDH1 showed the most significant and strongest downregulation after hsa-miR-142-3p, -17-5p, -223-3p, and -30e-5p transfection, respectively. The most significantly regulated gene set of each miRNA related to cell cycle (hsa-miRNA-144-3p and -5p [Padj = 4 × 10-40 and Padj = 4 × 10-6], -miR-17-5p [Padj = 9.5 × 10-23], -miR-30e-5p [Padj = 8.2 × 10-18], -miR-130a-3p [Padj = 5 × 10-15]), integrin cell surface interaction (-miR-223-3p [Padj = 2.4 × 10-7]), and interferon signaling (-miR-142-3p [Padj = 5 × 10-11]). At the end of acute inflammation, gingival miRNAs bring together complex regulatory networks that lead to increased expression of the gene MET. This underscores the importance of mesenchymal cell migration and invasion during gingival tissue remodeling and proliferation in restoring periodontal tissue homeostasis after active inflammation. MET, a receptor of the mitogenic hepatocyte growth factor fibroblast secreted, is a core gene of this process.

中文翻译：

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发炎牙龈中的 miRNA 将基因信号转导至 MET 表达增加。

几项基于阵列的 microRNA (miRNA) 表达研究独立显示 miRNA hsa-miR-130a-3p、-142-3p、-144-3p、-144-5p、-223-3p、-17-5p 和受牙周炎症影响的牙龈中的-30e-5p。我们的目的是确定这些 miRNA 调节的直接靶基因和信号通路，以确定与牙龈炎症反应和组织稳态相关的过程。我们将 7 种 miRNA 中每一种的 miRNA 模拟物 (mirVana) 分别转染到从 3 个不同供体培养的人原代牙龈成纤维细胞中。RNA测序后，进行差异基因表达和第二代基因集富集分析。使用定量逆转录酶聚合酶链式反应、蛋白质印迹和报告基因测定在转录物和蛋白质水平上验证 miRNA 的抑制和上调。所有 7 个 miRNA 均显着增加了 MET 原癌基因、受体酪氨酸激酶 (MET) 的表达。已知的牙周炎风险基因 CPEB1、ABCA1 和 ATP6V1C1 的表达分别被 hsa-miR-130a-3p、-144-3p 和 -144-5p 显着抑制。基因 WASL、ENPP5、ARL6IP1 和 IDH1 在 hsa-miR-142-3p、-17-5p、-223-3p 和 -30e-5p 转染后分别表现出最显着和最强的下调。与细胞周期相关的每个 miRNA 中最显着调控的基因集（hsa-miRNA-144-3p 和 -5p [Padj = 4 × 10-40 和 Padj = 4 × 10-6]、-miR-17-5p [Padj = 9.5 × 10-23]，-miR-30e-5p [Padj = 8.2 × 10-18]，-miR-130a-3p [Padj = 5 × 10-15]），整合素细胞表面相互作用（-miR-223） -3p [Padj = 2.4 × 10-7]）和干扰素信号传导（-miR-142-3p [Padj = 5 × 10-11]）。在急性炎症结束时，牙龈 miRNA 汇集了复杂的调控网络，导致 MET 基因表达增加。这强调了牙龈组织重塑和增殖过程中间充质细胞迁移和侵袭对于活动性炎症后恢复牙周组织稳态的重要性。MET是成纤维细胞分泌的促有丝分裂肝细胞生长因子的受体，是该过程的核心基因。