The Role of ADAR1 and ADAR2 in the Regulation of miRNA-21 in Idiopathic Pulmonary Fibrosis
The Role of ADAR1 and ADAR2 in the Regulation of miRNA-21 in Idiopathic Pulmonary Fibrosis
复制标题
DOI：
10.1007/s00408-018-0115-9
复制
发表时间：
2018-08-01
期刊：
影响因子：
5
通讯作者：
Ruiz, Victor
中科院分区：
文献类型：
--
作者：
Diaz-Pina, Gabriela;Ordonez-Razo, Rosa Ma.;Ruiz, Victor
microRNA (miRNA) 是小型非编码 1RNA，可在转录后调节基因表达。最近的证据表明作用于 RNA 的腺苷脱氨酶 (ADAR) 可以编辑 miRNA。 miRNA 参与不同疾病的发展，例如特发性肺纤维化 (IPF)。在 IPF 中，大约 40% 的 miRNA 与对照相比有差异表达。在这些 miRNA 中，miRNA-21 被发现在 IPF 中过度表达，其靶标是抗纤维化分子，例如 PELI1 和 SPRY2。本研究的目的是通过定量 ADAR1 和 2 的基因表达、蛋白水平以及过表达来确定 ADAR1 和 2 对人肺成纤维细胞中 miRNA-21 表达的作用。使用六个对照和六个纤维化原代成纤维细胞培养物进行 RNA 提取，测量 ADAR1、ADAR2、PELI1、SPRY2、miRNA-21 和 pri-miRNA-21 的表达。随后，使用两种纤维化成纤维细胞培养物过度表达ADAR1和ADAR2，并用TGFβ1刺激它们。进行实时PCR和Western印迹。ADAR1在IPF成纤维细胞中显着下调； ADAR1和ADAR2的过度表达重建了IPF患者成纤维细胞中miRNA-21、PELI1和SPRY2的表达水平。这些miRNA加工的变化在包括肺部疾病在内的病理诊断中具有重要价值，并且在理解不同病理发展的分子机制以及代表新的治疗靶点方面发挥着重要作用。
microRNAs (miRNAs) are small non-coding 1RNAs that post-transcriptionally regulate gene expression. Recent evidence shows that adenosine deaminases that act on RNA (ADAR) can edit miRNAs. miRNAs are involved in the development of different diseases, such as idiopathic pulmonary fibrosis (IPF). In IPF, about 40% of the miRNAs are differentially expressed with respect to controls. Among these miRNAs, miRNA-21 has been found over-expressed in IPF and its targets are anti-fibrosing molecules such as PELI1 and SPRY2. The objective of this study is to determine the role of ADAR1 and 2 on the expression of miRNA-21 in human lung fibroblasts trough quantification of gene expression, protein levels, and overexpression of ADAR1 and 2.Six control and six fibrotic primary fibroblast cell cultures were used for RNA extraction, ADAR1, ADAR2, PELI1, SPRY2, miRNA-21, and pri-miRNA-21 expression was measured. Subsequently, two fibrotic fibroblast cultures were used for overexpression of ADAR1 and ADAR2, and they were stimulated with TGF beta 1. Real-time PCR and Western blot were performed.ADAR1 is significantly downregulated in IPF fibroblasts; the overexpression of ADAR1 and ADAR2 reestablishes the expression levels of miRNA-21, PELI1, and SPRY2 in fibroblasts of patients with IPF.These changes in the processing of miRNAs have great value in pathology diagnosis, including lung diseases, and play an important role in the understanding of molecular mechanisms involved in the development of different pathologies, as well as representing new therapeutic targets.