MicroRNA-132 targeting PTEN contributes to cilostazol-promoted vascular smooth muscle cell differentiation
MicroRNA-132 targeting PTEN contributes to cilostazol-promoted vascular smooth muscle cell differentiation
复制标题
DOI：
10.1016/j.atherosclerosis.2018.04.030
复制
发表时间：
2018-07-01
期刊：
影响因子：
5.3
通讯作者：
Yeh, Yung-Hsin
中科院分区：
文献类型：
--
作者：
Chen, Wei-Jan;Chen, Ying-Hwa;Yeh, Yung-Hsin
背景与目的：西洛他唑除具有抗血小板作用外，还能促进血管平滑肌细胞(VSMC)分化。本研究的目的是探讨已知与VSMC分化相关的PTEN及其相关的microRNA(MiRNA)在西洛他唑依赖效应中的潜在作用。方法和结果：通过对球囊损伤的大鼠颈动脉进行比较，发现miR-132在球囊损伤前后的表达存在差异。生物信息学分析预测PTEN将成为miR-132的新靶点。Western印迹、实时定量逆转录-聚合酶链式反应和原位杂交结果表明，西洛他唑治疗使球囊损伤动脉新生内膜PTEN表达增强，miR-132表达降低。用西洛他唑处理培养的大鼠VSMCs后，PTEN mRNA表达上调，miR-132表达下调，这与体外培养的VSMCs的作用有关。共转染实验表明，将miR-132模拟物导入VSMC后，抑制了PTEN 3‘非编码区的活性，进一步表明PTEN是miR-132的直接靶点。在VSMC中过表达miR-132可抑制西洛他唑诱导的PTEN及其下游VSMC分化标志物(Calponin)的表达，证实了miR-132在VSMC分化中的关键作用。瞬时转染研究表明，西洛他唑可降低miR-132启动子的活性，这种作用是通过环AMP反应元件结合蛋白介导的。结论：靶向PTEN的miR-132可能是介导西洛他唑诱导VSMC分化的重要调节因子。(C)2018爱思唯尔B.V.保留所有权利。
Background and aims: Cilostazol, beyond its antiplatelet effect, is also capable of promoting vascular smooth muscle cell (VSMC) differentiation. The aim of this study was to explore the potential role of PTEN, known to associate with VSMC differentiation, and its related microRNA (miRNA) in cilostazol-dependent effects.Methods and results: Microarray analysis in balloon-injured rat carotid arteries comparing with and without balloon injury revealed that miR-132 was differentially expressed. Bioinformatic analysis predicts PTEN as a novel target of miR-132. Western blot and quantitative real-time reverse transcription-polymerase chain reaction along with in situ hybridization documented that cilostazol treatment enhanced PTEN and reduced miR-132 expression in the neointima of balloon-injured arteries. Treatment of cultured rat VSMCs with cilostazol resulted in the up-regulation of PTEN mRNA and the down-regulation of miR-132, supporting an in vitro relevance. Co-transfection experiments showed that transfection of miR-132 mimic into VSMCs suppressed PTEN 3'UTR activities, further reflecting that PTEN is the direct target of miR-132. Over-expression of miR-132 in VSMCs led to an attenuation of cilostazol-induced PTEN and its downstream VSMC differentiation marker (calponin) expression, confirming the critical role of miR-132 in VSMC differentiation. Transient transfection studies demonstrated that cilostazol reduced the activity of miR-132 promoter, which was mediated via cyclic AMP response element-binding protein. Notably, the use of lentivirus to over-express miR-132 in the neointima of balloon-injured arteries could reverse the effect of cilostazol in vivo.Conclusions: These results suggest that miR-132 by targeting PTEN may be an important regulator in mediating cilostazol actions on VSMC differentiation. (C) 2018 Elsevier B.V. All rights reserved.