Dysregulated miR-124 and miR-200 expression contribute to cholangiocyte proliferation in the cholestatic liver by targeting IL-6/STAT3 signalling
Dysregulated miR-124 and miR-200 expression contribute to cholangiocyte proliferation in the cholestatic liver by targeting IL-6/STAT3 signalling
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DOI：
10.1016/j.jhep.2014.10.033
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发表时间：
2015-04-01
影响因子：
25.7
通讯作者：
Cai, Wei
中科院分区：
文献类型：
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作者：
Xiao, Yongtao;Wang, Jun;Cai, Wei
背景与目的：胆汁淤积性肝病与microRNAs （miRNAs）表达异常有关。然而，目前尚不清楚mirna是否参与胆汁淤积诱导的胆管细胞增殖。在这项研究中，我们验证了mirna通过影响IL-6途径调节胆管细胞增殖的假设，IL-6途径是胆管细胞增殖的已知调节剂。方法：观察胆道闭锁（BA）患者和胆管结扎（BDL）大鼠IL-6、Foxa2和磷酸化信号转导激活因子3 （STAT3）的表达。采用miRNA阵列和实时荧光定量PCR检测BA患者和BDL大鼠的miRNA表达。采用免疫印迹法、免疫组化法和增殖法研究mirna的生物学功能。荧光素酶报告基因检测和Western blot检测miRNA靶标。结果：与对照组相比，BA患者和BDL大鼠肝脏白细胞介素-6 （IL-6）表达显著升高，miR-124表达显著降低。此外，STAT3和IL-6受体（IL-6R）的mRNA水平与miR-124呈负相关。miR-124的异位表达通过直接靶向STAT3和IL-6R的3'-非翻译区，抑制il -6介导的体外胆管细胞增殖和体内胆管细胞增生。我们进一步证明miR-200家族成员在胆汁淤淤中显著上调，抑制胆管细胞中FOXA2的表达，从而进一步增强il的表达。结论：我们的研究结果表明，miR-124的下调和miR-200的上调通过IL-6/STAT3途径协同促进胆管增殖。(C) 2014欧洲肝脏研究协会。Elsevier B.V.版权所有。
Background & Aims: Cholestatic liver disease is associated with dysregulated expression of microRNAs (miRNAs). However, it remains unknown whether miRNAs are involved in the cholestasis-induced proliferation of cholangiocytes. In this study, we tested the hypothesis that miRNAs modulate cholangiocyte proliferation through effects on the IL-6 pathway, a known regulator of cholangiocyte proliferation.Methods: Expression of IL-6, Foxa2, and phosphorylated signal transducer activator of transcription 3 (STAT3) was investigated in patients with biliary atresia (BA) and in rats subjected to bile duct ligation (BDL). miRNA expression was determined in BA patients and BDL rats, with miRNA array and quantitative real-time PCR. Biological functions of miRNAs were studied using immunoblot, immunohistochemical and proliferation assays. Luciferase reporter assays and Western blots were performed to identify miRNA targets.Results: Hepatic interleukin-6 (IL-6) expression was significantly elevated in BA patients and BDL rats, while the expression of miR-124 was dramatically decreased in comparison to controls. Moreover, mRNA levels of STAT3 and IL-6 receptor (IL-6R) were inversely correlated with those of miR-124. Ectopic expression of miR-124 inhibited IL-6-mediated cholangiocyte proliferation in vitro and cholangiocyte hyperplasia in vivo, through a mechanism involving direct targeting of the 3'-untranslated region of STAT3 and IL-6R. We further demonstrated that miR-200 family members were significantly upregulated in cholestasis and inhibited FOXA2 expression in cholangiocytes, which further enhanced the expression of IL-Conclusions: Our findings suggest that downregulation of miR-124 and upregulation of miR-200 collaboratively promote bile duct proliferation through the IL-6/STAT3 pathway. (C) 2014 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.