Sixth Medical Department, Hunan Chest Hospital, Changsha, Hunan 410013, P.R. China.
Endoscopy Center, Hunan Chest Hospital, Changsha, Hunan 410013, P.R. China.
Int J Mol Med. 2021 May;47(5). doi: 10.3892/ijmm.2021.4913. Epub 2021 Mar 24.
Idiopathic pulmonary fibrosis (IPF) is the most common form of idiopathic interstitial pneumonia with an increasing incidence. In the present study, Genome Expression Omnibus (GEO) datasets (GSE10667, GSE24206 and GSE32537) were applied to identify lncRNA DLEU2 in IPF. Through prediction using starBase, TargetScan, miRTarBase and miRDB, tripartite motif containing 2 (TRIM2) and prostaglandin F2 receptor inhibitor (PTGFRN) were found to be upregulated in IPF. DLEU2 expression, the mRNA expression of TRIM2 and PTGFRN, and miR‑369‑3p expression in A549 cells and lung tissues were detected by RT‑qPCR. The protein expression of TRIM2 and PTGFRN in lung tissues and A549 cells was detected by western blot analysis. The proliferation and migration of A549 cells was respectively detected by CCK‑8 assay and wound healing assay. The expression of collagen I, α‑smooth muscle actin (SMA) and E‑cadherin was detected by immunofluorescence assay in A549 cells, and collagen I expression was detected by immunohistochemistry assay in lung tissues. The expression of collagen I, α‑SMA and E‑cadherin was also detected by western blot analysis in A549 cells and lung tissues. Dual‑luciferase reporter assay was used to confirm the association between DLEU2 and miR‑369‑3p, and miR‑369‑3p and TRIM2. As a result, DLEU2 expression was found to be upregulated in IPF and in transforming growth factor (TGF)‑β1‑stimulated A549 cells. The silencing of DLEU2 inhibited the TGF‑β1‑induced proliferation, migration and epithelial‑mesenchymal transition (EMT) of A549 cells and bleomycin (BLM)‑induced pulmonary fibrosis in mice. TRIM2 expression was increased and miR‑369‑3p expression was decreased in the lung tissues of mice with BLM‑induced fibrosis and in TGF‑β1‑stimulated A549 cells. DLEU2 directly targeted miR‑369‑3p. The effect of the silencing of DLEU2 on TGF‑β1‑stimulated A549 cells was suppressed by the silencing of miR‑369‑3p. TRIM2 was the target protein of miR‑369‑3p. On the whole, the present study demonstrates that the silencing of DLEU2 suppressed IPF by upregulating miR‑369‑3p expression and downregulating TRIM2 expression.
特发性肺纤维化(IPF)是特发性间质性肺炎中最常见的一种,发病率呈上升趋势。本研究应用基因组表达综合数据库(GEO)数据集(GSE10667、GSE24206 和 GSE32537)鉴定出 IPF 中的 lncRNA DLEU2。通过 starBase、TargetScan、miRTarBase 和 miRDB 的预测,发现三肽基含 2 结构域蛋白 2(TRIM2)和前列腺素 F2 受体抑制剂(PTGFRN)在 IPF 中上调。通过 RT-qPCR 检测 A549 细胞和肺组织中的 DLEU2 表达、TRIM2 和 PTGFRN 的 mRNA 表达以及 miR-369-3p 的表达。通过 Western blot 分析检测肺组织和 A549 细胞中 TRIM2 和 PTGFRN 的蛋白表达。通过 CCK-8 测定法和划痕愈合测定法分别检测 A549 细胞的增殖和迁移。通过免疫荧光法检测 A549 细胞中胶原 I、α-平滑肌肌动蛋白(SMA)和 E-钙黏蛋白的表达,并通过免疫组织化学法检测肺组织中胶原 I 的表达。通过 Western blot 分析检测 A549 细胞和肺组织中胶原 I、α-SMA 和 E-钙黏蛋白的表达。双荧光素酶报告基因测定法用于证实 DLEU2 与 miR-369-3p 以及 miR-369-3p 与 TRIM2 之间的关联。结果发现,IPF 患者和转化生长因子(TGF)-β1 刺激的 A549 细胞中 DLEU2 的表达上调。DLEU2 的沉默抑制了 TGF-β1 诱导的 A549 细胞增殖、迁移和上皮-间充质转化(EMT)以及博来霉素(BLM)诱导的小鼠肺纤维化。BLM 诱导的纤维化小鼠和 TGF-β1 刺激的 A549 细胞中 TRIM2 表达增加,miR-369-3p 表达降低。DLEU2 可直接靶向 miR-369-3p。沉默 miR-369-3p 可抑制 DLEU2 沉默对 TGF-β1 刺激的 A549 细胞的影响。TRIM2 是 miR-369-3p 的靶蛋白。总的来说,本研究表明,沉默 DLEU2 通过上调 miR-369-3p 的表达和下调 TRIM2 的表达来抑制 IPF。