MicroRNA-221-3p promotes pulmonary artery smooth muscle cells proliferation by targeting AXIN2 during pulmonary arterial hypertension
MicroRNA-221-3p promotes pulmonary artery smooth muscle cells proliferation by targeting AXIN2 during pulmonary arterial hypertension
复制标题
MicroRNA-221-3p 在肺动脉高压期间通过靶向 AXIN2 促进肺动脉平滑肌细胞增殖
DOI：
10.1016/j.vph.2017.07.002
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发表时间：
2019-05-01
影响因子：
4
通讯作者：
Chen, Jingyu
中科院分区：
文献类型：
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作者：
Nie, Xiaowei;Chen, Yuan;Chen, Jingyu
肺动脉高压（PAH）是一种以肺动脉平滑肌细胞（PASMC）过度增殖和迁移为特征的病理状态。多环芳烃的发病机制与细胞过度增殖和细胞凋亡抵抗等癌症有相似之处。本课课组之前的一项研究表明，肿瘤抑制因子axin2（轴抑制蛋白2）的表达降低是PASMC增殖增强和细胞凋亡抑制的原因。然而，调控PAH中AXIN2下调的机制仍不明确。本研究的数据表明，miR-221-3p作为AXIN2的上游调节因子，具有诱导PASMC增殖的功能。我们首先发现miR-221-3p在PAH患者的肺组织和PASMC以及PAH动物模型中表达升高。分别用miR-221-3p模拟物和miR-221-3p抑制剂转染人PASMC，并通过BrdU掺入、PCNA染色和伤口愈合试验评估其对增殖和迁移的影响。此外，我们研究了miR-221-3p在PASMC中促进细胞增殖的分子机制，并通过双荧光素酶报告基因检测、qRT-qPCR和western blotting鉴定出AXIN2是miR-221-3p的直接靶基因。此外，我们发现XAV-939异位表达AXIN2或药理抑制β -catenin可以减弱miR-221-3p对PASMC细胞增殖的影响。此外，静脉注射miR-221-3p抑制剂可以减轻su5416 -缺氧诱导的大鼠PAH的进展。本研究的结果确定了miR-221-3p和AXIN2调节PASMC增殖的新调控轴。
Pulmonary arterial hypertension (PAH) is a pathological condition characterized by excessive cell proliferation and migration of pulmonary arterial smooth muscle cells (PASMC). PAH pathogenesis shares similarities with cancers such as excessive cell proliferation and apoptosis resistance. A previous study by our group revealed that decreased expression of a tumor suppressor-AXIN2 (Axis inhibition protein 2) was responsible for enhanced PASMC proliferation and suppressed apoptosis. Nevertheless, the mechanisms that regulate the downregulation of AXIN2 in PAH remain elusive. Data from the present study demonstrated that miR-221-3p acts as an upstream regulator of AXIN2 and functions to induce PASMC proliferation. We first showed that miR-221-3p expression was elevated in lung tissue and PASMC of PAH patients as well as in animal models of PAH. Human PASMC were transfected with a miR-221-3p mimic and miR-221-3p inhibitor, respectively, and their effects on the proliferation and migration was assessed using BrdU incorporation, PCNA staining and wound healing assays. In addition, we investigated the molecular mechanism through which miR-221-3p contributes to cell proliferation in PASMC and identified AXIN2 as a direct target gene of miR-221-3p by dual luciferase reporter gene assays, qRT-qPCR and western blotting. Furthermore, we found that ectopic expression of AXIN2 or pharmacological inhibition of beta-catenin by XAV-939 can attenuate the effect of miR-221-3p on cell proliferation in PASMC. Moreover, intravenous injection of miR-221-3p inhibitor attenuated the progression of SU5416-hypoxia-induced PAH in rats. The results of the present study identified a new regulatory axis in which miR-221-3p and AXIN2 regulate the proliferation of PASMC.