MicroRNA-221/222 upregulation indicates the activation of stellate cells and the progression of liver fibrosis
MicroRNA-221/222 upregulation indicates the activation of stellate cells and the progression of liver fibrosis
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DOI：
10.1136/gutjnl-2011-300717
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发表时间：
2012-11-01
期刊：
影响因子：
24.5
通讯作者：
Kawada, Norifumi
中科院分区：
文献类型：
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作者：
Ogawa, Tomohiro;Enomoto, Masaru;Kawada, Norifumi
背景微小RNA(miRNA)在肝脏病理生理学和肝癌的发生发展中具有重要作用。目的探索在慢性肝病引起的肝纤维化进展中受调控的miRNA。设计通过微阵列分析鉴定感染丙型肝炎病毒的人肝脏中受调控的miRNA。通过实时 RT-PCR 验证了它们在患有非酒精性脂肪性肝炎的人类肝脏、来自两种纤维化模型的小鼠肝脏和培养的星状细胞中的表达。测定了核因子 kappa B (NF-kappa B) 对星状细胞中 miR-222 表达的调节。最后，确定miR-222前体或抑制剂对细胞周期蛋白依赖性激酶抑制剂1B（CDKN1B）表达和LX-2细胞生长的影响。结果发现，miR-199a-5p/199a-3p和miR-221/222在人肝脏中以纤维化进展依赖性方式上调。在这些 miRNA 中，miR-221/222 在 LX-2 细胞中上调，并在小鼠原代星状细胞的培养依赖性激活过程中增加，其方式类似于 α1(I) 胶原蛋白和 α-平滑肌肌动蛋白 mRNA 的表达。肝纤维化小鼠模型中 miR-221/222 的表达增加。相比之下，NF-κ B 抑制剂显着抑制了培养物中通过转化生长因子 α 或肿瘤坏死因子 α 刺激的 miR-222 诱导。虽然miR-222的过表达或下调未能调节LX-2细胞的生长，但miR-222与CDKN1B 3'UTR结合并调节相应蛋白的表达。 结论 miR-221/222可能是星状细胞活化和肝纤维化进展的新标志物。
Background MicroRNAs (miRNAs) are important in hepatic pathophysiology and the development of liver cancer.Objective To explore miRNAs that are regulated with the progression of liver fibrosis caused by chronic liver disease.Design The regulated miRNAs in human livers infected with hepatitis C virus were identified by microarray analysis. Their expression in human livers with nonalcoholic steatohepatitis, mouse livers from two fibrosis models and cultured stellate cells was validated by realtime RT-PCR. The regulation of miR-222 expression in stellate cells by nuclear factor kappa B (NF-kappa B) was assayed. Finally, the effects of an miR-222 precursor or inhibitor on the expression of cyclin-dependent kinase inhibitor 1B (CDKN1B) and the growth of LX-2 cells were determined.Results It was found that miR-199a-5p/199a-3p and miR-221/222 were upregulated in the human liver in a fibrosis progressionedependent manner. Among these miRNAs, miR-221/222 were upregulated in LX-2 cells and increased during the course of culture-dependent activation of mouse primary stellate cells, in a manner similar to the expression of alpha 1(I) collagen and alpha-smooth muscle actin mRNAs. The expression of miR-221/222 increased in mouse models of liver fibrosis. In contrast, an NF-kappa B inhibitor significantly suppressed the miR-222 induction that was stimulated in culture by transforming growth factor alpha or tumour necrosis factor alpha. Although overexpression or downregulation of miR-222 failed to regulate the growth of LX-2 cells, miR-222 bound to the CDKN1B 3'UTR and regulated the expression of the corresponding protein.Conclusion miR-221/222 may be new markers for stellate cell activation and liver fibrosis progression.