MicroRNA-221 promotes proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs) by targeting tissue inhibitor of metalloproteinases-3 (TIMP3)
MicroRNA-221 promotes proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs) by targeting tissue inhibitor of metalloproteinases-3 (TIMP3)
复制标题
MicroRNA-221 通过靶向金属蛋白酶-3 (TIMP3) 组织抑制剂促进肺动脉平滑肌细胞 (PASMC) 的增殖和迁移
DOI：
10.21037/cdt-20-328
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发表时间：
2020-08-01
影响因子：
2.4
通讯作者：
Zhang, Haifeng
中科院分区：
文献类型：
--
作者：
Yan, Yan;Xu, Ying;Zhang, Haifeng
背景资料：血管平滑肌细胞（VSMC）的异常增殖和迁移在包括肺动脉高压（PAH）在内的心血管疾病的发生发展中起着重要作用。microRNA（miRNAs，miRs）被认为与PAH发病机制的进展有关。本研究旨在阐明miR-221在肺动脉平滑肌细胞（PASMCs）增殖和迁移中的作用，并寻找参与这一生物学过程的靶基因。5-乙炔基-2 '-脱氧尿苷（EdU）法检测细胞增殖;通过划痕试验测定细胞迁移。实时定量PCR检测miR-221的表达，Western blot检测TIMP 3的表达。荧光素酶法证实TIMP 3是miR-221的直接靶基因。建立野百合碱（MCT）诱导的PAH大鼠模型，检测肺组织和PASMC中miR-221和TIMP 3的表达。miR-221在PASMCs中对TIMP 3表达有负调控作用。此外，基于荧光素酶测定，TIMP 3被鉴定为PASMCs中miR-221的直接靶基因。TIMP 3敲低可消除miR-221抑制剂对PASMCs增殖和迁移的抑制作用，提示TIMP 3介导miR-221在PASMCs中的作用。结论：miR-221通过靶向TIMP 3促进PASMCs增殖和迁移。miR-221和TIMP 3可能是治疗PAH的潜在靶点。
Background: Aberrant vascular smooth muscle cell (VSMC) proliferation and migration play an important role in the development of cardiovascular diseases including pulmonary arterial hypertension (PAH). MicroRNAs (miRNAs, miRs) have been considered to be implicated in the progression of PAH pathogenesis. In this study, we aim to clarify the role of miR-221 on proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs) and identify the target genes involved in this biological process.Methods: PASMCs were isolated from the pulmonary arteries of male Sprague-Dawley (SD) rats. Cell proliferation of PASMCs was detected by 5-ethynyl-2'-deoxyuridine (EdU) assay. Cell migration was determined by a scratch wound assay. Quantitative real-time PCR was used to determine the expression of miR-221 while western blot analysis was used to determine the expression of TIMP3. Luciferase assay was used to confirm that TIMP3 was a direct target gene of miR-221. Monocrotaline (MCT) induced-PAH rat model was established and miR-221 and TIMP3 levels were checked in lung tissue and PASMCs from PAH rats.Results: miR-221 was able to promote the proliferation and migration PASMCs. TIMP3 were negatively regulated by miR-221 at the protein level in PASMCs. In addition, TIMP3 was identified to be a direct target gene of miR-221 in PASMCs based on luciferase assays. TIMP3 knockdown abolished the inhibitory effect of rniR-221 inhibitor on PASMCs proliferation and migration, suggesting TIMP3 mediated the effects of miR-221 in PASMCs. Finally, we found that miR-221 was increased while TLMP3 was down-regulated in PASMCs in MCT-treated rats.Conclusions: In conclusion, miR-221 promotes PASMCs proliferation and migration by targeting TIMP3. MiR-221 and TIMP3 could be potential therapeutic targets for the treatment of PAH.