Mitomycin C induces fibroblasts apoptosis and reduces epidural fibrosis by regulating miR-200b and its targeting of RhoE
Mitomycin C induces fibroblasts apoptosis and reduces epidural fibrosis by regulating miR-200b and its targeting of RhoE
复制标题
Mitomycin C 通过调节 miR-200b 及其对 RhoE 的靶向作用，诱导成纤维细胞凋亡并减少硬膜外纤维化。
DOI：
10.1016/j.ejphar.2015.08.002
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发表时间：
2015-10-15
影响因子：
5
通讯作者：
Lu, Chun
中科院分区：
文献类型：
--
作者：
Sun, Yu;Ge, Yingbin;Lu, Chun
丝裂霉素C（MMC）已知可减少硬膜外纤维化，但其潜在机制尚未阐明。miR-200b的异常表达在多种疾病的多种纤维化组织中均有报道。本研究的目的是阐明MMC诱导成纤维细胞凋亡和减轻硬膜外纤维化的机制。MMC处理后测定人成纤维细胞中miR-200 b的表达，并使用荧光素酶活性测定法测定miR-200 b和RhoE之间的靶向关联。采用流式细胞术和western blot分析评价MMC和miR-200 b对人成纤维细胞凋亡的影响。通过Rydell分级、羟脯氨酸含量、凋亡细胞计数和组织学分析评价MMC和miR-200 b对硬膜外纤维化的影响。MMC可显著下调miR-200 b的表达，诱导人成纤维细胞凋亡。miR-200 b的直接下调可诱导人成纤维细胞凋亡。此外，我们在RhoE的3 '非翻译区内鉴定了miR-200b的结合序列。RhoE被证实是miR-200b的直接靶点，RhoE本身作为成纤维细胞凋亡的促进剂。抑制miR-200 b可增加大鼠成纤维细胞凋亡，减轻硬膜外纤维化，这与MMC的作用一致。本研究提示MMC通过调控miR-200 b的表达及其对RhoE的靶向作用，诱导成纤维细胞凋亡，减轻硬膜外纤维化。(C)2015爱思唯尔B.V.保留所有权利。
Mitomycin C (MMC) is known to reduce epidural fibrosis, but the underlying mechanisms have not yet been elucidated. Aberrant miR-200b expressions have been reported in multiple types of fibrotic tissues from many diseases. The aim of this study was to clarify the mechanism by which MMC induces fibroblasts apoptosis and reduces epidural fibrosis. The expression of miR-200b in human fibroblasts was determined after MMC treatment, and the targeted association between miR-200b and RhoE was determined using the luciferase activity assay. The effects of MMC and miR-200b on human fibroblasts apoptosis were evaluated using flow cytometry and western blot analysis. The effects of MMC and miR-200b on epidural fibrosis were evaluated using the Rydell classification, hydroxyproline content, apoptotic cell count and histological analysis. The study revealed that MMC could significantly downregulate miR-200b expression and induce human fibroblasts apoptosis. The direct downregulation of miR-200b could induce human fibroblasts apoptosis. Furthermore, we identified the binding sequence for miR-200b within the 3' untranslated region of RhoE. RhoE was confirmed to be a direct target of miR-200b, and RhoE itself acted as a promoter of fibroblasts apoptosis. The inhibition of miR-200b increased fibroblasts apoptosis and reduced epidural fibrosis in rats, which was in accordance with the effect of MMC. This study suggests that MMC induces fibroblasts apoptosis and reduces epidural fibrosis by regulating miR-200b expression and its targeting of RhoE. (C) 2015 Elsevier B.V. All rights reserved.