LncRNA Mical2/miR-203a-3p sponge participates in epithelial-mesenchymal transition by targeting p66Shc in liver fibrosis.
LncRNA Mical2/miR-203a-3p sponge participates in epithelial-mesenchymal transition by targeting p66Shc in liver fibrosis.
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DOI：
10.1016/j.taap.2020.115125
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发表时间：
2020-07
影响因子：
3.8
通讯作者：
Yan Zhao;Zhecheng Wang;Junjun Zhou;Dongcheng Feng;Yang Li;Yan Hu;Feng Zhang;Zhao Chen;
中科院分区：
文献类型：
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作者：
Yan Zhao;Zhecheng Wang;Junjun Zhou;Dongcheng Feng;Yang Li;Yan Hu;Feng Zhang;Zhao Chen;
肝纤维化中上皮-间质转化（EMT）受活性氧（ROS）调节。p66 Shc是一种氧化还原酶，但其在肝纤维化中EMT的作用尚不清楚。长链非编码RNA（longnoncodingRNA，lncRNA）在许多生理和病理过程中起着重要的调节作用，通常作为microRNA（microRNA，miRNA）海绵来调节基因表达。本研究旨在探讨p66 Shc在肝纤维化EMT中的作用及lncRNA海绵对p66 Shc的调控作用。在体内，p66 Shc沉默阻止四氯化碳（CCl 4）诱导的EMT，如通过上调E-钙粘蛋白，下调波形蛋白和N-钙粘蛋白，抑制氧化应激和细胞外基质（ECM）成分所证明的。此外，在体外，TGF-β1显着增强ECM成分，以及EMT表型的发展。p66 Shc下调可消除这些效应，而p66 Shc过表达可加重这些效应。从机制上讲，p66 Shc通过介导ROS促进EMT，如在外源性过氧化氢（H2 O2）刺激下p66 Shc下调抑制EMT所证明的。此外，我们发现与CasL 2（Mical 2）lncRNA相互作用的分子作为内源性miR-203 a-3 p海绵来调节p66 Shc表达。Mical 2沉默和miR-203 a-3 p agomiR处理均下调p66 Shc表达，从而在体内和体外抑制EMT。值得注意的是，鼠纤维化中增加的p66 Shc和Mical 2水平和降低的miR-203 a-3 p水平与肝纤维化患者中的那些一致。总之，我们的研究揭示了p66 Shc在肝纤维化中是关键的，并且Mical 2、miR-203 a-3 p和p66 Shc构成了肝纤维化中的新的调节途径。
Epithelial-mesenchymal transition (EMT) is regulated by reactive oxygen species (ROS) in liver fibrosis. p66Shc is a redox enzyme, but its role of EMT is unclear in liver fibrosis. Long noncoding RNAs (lncRNAs) have been implicated as important regulators in numerous physiological and pathological processes and generally acting as a microRNA (miRNA) sponge to regulate gene expression. The aim of the current study was to evaluate the contribution of p66Shc to EMT in liver fibrosis and the regulation of p66Shc by lncRNA sponge. In vivo, p66Shc silencing prevented carbon tetrachloride (CCl4)-induced EMT as evidenced by the upregulation of E-cadherin, downregulation of Vimentin and N-cadherin, and inhibition of oxidative stress and extracellular matrix (ECM) components. Moreover, in vitro, TGF-β1 significantly enhanced ECM components, as well as the development of the EMT phenotype. These effects were abrogated by p66Shc downregulation and aggravated by p66Shc overexpression. Mechanistically, p66Shc contributed to EMT via mediating ROS, as evidenced by p66Shc downregulation inhibiting EMT under exogenous hydrogen peroxide (H2O2) stimulation. Furthermore, we found that molecule interacting with CasL2 (Mical2) lncRNA functioned as an endogenous miR-203a-3p sponge to regulate p66Shc expression. Both Mical2 silencing and miR-203a-3p agomiR treatment downregulated p66Shc expression, thus suppressing EMT in vivo and in vitro. Notably, the increased p66Shc and Mical2 levels and decreased miR-203a-3p levels in murine fibrosis were consistent with those in patients with liver fibrosis. In sum, our study reveals that p66Shc is critical for liver fibrosis and that Mical2, miR-203a-3p and p66Shc compose a novel regulatory pathway in liver fibrosis.