MicroRNA-30a Suppresses the Activation of Hepatic Stellate Cells by Inhibiting Epithelial-to-Mesenchymal Transition
MicroRNA-30a Suppresses the Activation of Hepatic Stellate Cells by Inhibiting Epithelial-to-Mesenchymal Transition
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MicroRNA-30a 通过抑制上皮间质转化来抑制肝星状细胞的活化
DOI：
10.1159/000488411
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Zhou, Mengtao
中科院分区：
文献类型：
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作者：
Zheng, Jianjian;Wang, Wei;Zhou, Mengtao
背景/目的：肝星状细胞（HSCs）的激活被认为是肝纤维化的关键事件，已有报道称上皮-间质转化（EMT）过程参与了肝星状细胞的激活。众所周知，microRNAs （miRNAs）在HSC活化中起促纤维化或抗纤维化作用。最近，新的研究表明，miR-30a在人类癌症中下调，过表达miR-30a通过抑制EMT过程抑制肿瘤生长和侵袭。然而，miR-30a是否能够调控HSC激活中的EMT过程尚不清楚。方法：采用real-time PCR方法定量检测四氯化碳（CCl4）诱导的大鼠肝纤维化、活化的hsc和肝硬化患者中miR-30a的表达。我们还分析了miR-30a在体内和体外肝纤维化中的作用。荧光素酶活性测定检测了miR-30a与蜗牛家族转录抑制因子1 (Snai1) 3 ' -未翻译区域的结合。结果：miR-30a在人肝硬化组织中下调。在CCl4大鼠中，在纤维化肝组织和分离的hsc中发现了miR-30a的降低。在培养期间，原代造血干细胞中miR-30a水平显著降低。miR-30a过表达可抑制ccl4诱导的肝纤维化。恢复miR-30a可抑制HSC活化，包括细胞增殖、α-SMA和胶原表达。值得注意的是，miR-30a抑制EMT过程，TGF-β1和Vimentin减少，GFAP和E-cadherin增加。与对照组相比，miR-30a诱导Snai1蛋白表达显著降低。有趣的是，在肝纤维化过程中，Snail蛋白表达增加，表明miR-30a水平与Snai1蛋白表达可能存在负相关。进一步的研究表明Snai1是miR-30a的靶点。结论：我们的研究结果表明，miR-30a至少在一定程度上通过降低Snai1抑制EMT过程，从而抑制肝纤维化中HSC的激活。
Background/Aims: The activation of hepatic stellate cells (HSCs) is considered as a pivotal event in liver fibrosis and epithelial-mesenchymal transition (EMT) process has been reported to be involved in HSC activation. It is known that microRNAs (miRNAs) play a pro-fibrotic or anti-fibrotic role in HSC activation. Recently, emerging studies show that miR-30a is down-regulated in human cancers and over-expression of miR-30a inhibits tumor growth and invasion via suppressing EMT process. However, whether miR-30a could regulate EMT process in HSC activation is still unclear. Methods: miR-30a expression was quantified using real-time PCR in carbon tetrachloride (CCl4)-induced rat liver fibrosis, activated HSCs and patients with cirrhosis. Roles of miR-30a in liver fibrosis in vivo and in vitro were also analyzed. Luciferase activity assays were performed to examine the binding of miR-30a to the 3′-untranslated region of snail family transcriptional repressor 1 (Snai1). Results: miR-30a was down-regulated in human cirrhotic tissues. In CCl4 rats, reduced miR-30a was found in fibrotic liver tissues as well as isolated HSCs. There was a significant reduction in miR-30a in primary HSCs during culture days. miR-30a over-expression resulted in the suppression of CCl4-induced liver fibrosis. Restoration of miR-30a led to the inhibition of HSC activation including cell proliferation, α-SMA and collagen expression. Notably, miR-30a inhibited EMT process, with a reduction in TGF-β1 and Vimentin as well as an increase in GFAP and E-cadherin. miR-30a induced a significant reduction in Snai1 protein expression when compared with the control. Interestingly, Snail protein expression was increased during liver fibrosis, indicating that there may be a negative correlation between miR-30a level and Snai1 protein expression. Further studies demonstrated that Snai1 was a target of miR-30a. Conclusion: Our results suggest that miR-30a inhibits EMT process, at least in part, via reduction of Snai1, leading to the suppression of HSC activation in liver fibrosis.