Molecular mechanism underlying miR‑130b‑Sp1 transcriptional regulation in LPS‑induced upregulation of MUC5AC in the bile duct epithelium.
Molecular mechanism underlying miR‑130b‑Sp1 transcriptional regulation in LPS‑induced upregulation of MUC5AC in the bile duct epithelium.
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DOI：
10.3892/mmr.2020.11745
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发表时间：
2021-03
影响因子：
3.4
通讯作者：
Wu S
中科院分区：
文献类型：
--
作者：
Wu X;Yao C;Kong J;Tian Y;Fan Y;Zhang Z;Han J;Wu S
肝内胆管结石是一种常见病，严重威胁着中国人群的健康。肝内胆管结石的病理机制与肝内胆管细菌感染、慢性炎症和粘蛋白5AC（MUC 5AC）过度表达密切相关。然而，负责脂多糖（LPS）诱导的MUC 5AC上调的确切机制尚未阐明。特异性蛋白1（Sp1）是一种普遍存在的转录因子，在调节许多负责正常细胞功能的基因中起着至关重要的作用。microRNA（miR/miRNA）-130b是miRNA家族的成员。miRNAs可与靶基因的3′-非翻译区（3′-UTR）结合，影响其表达水平。本研究发现LPS通过影响Sp1的分泌而增加MUC 5AC的表达。染色质免疫沉淀-定量PCR实验进一步验证了MUC 5AC启动子序列中的3个Sp1结合位点可以调控MUC 5AC的表达。进一步分析表明Sp1的表达受miR-130 b的调控。荧光素酶实验在Sp1 3′-UTR区域鉴定了一个miR-130 b结合位点。体内实验也证实了miR-130 b-Sp1-MUC 5AC信号通路在胆管结石形成中的作用，并表明该通路可能为肝内胆管结石的治疗提供靶向治疗策略。
Hepatolithiasis is a common disease that represents a serious health threat to the Chinese population. The pathological mechanism underlying hepatolithiasis is closely related to bacterial infections of the intrahepatic bile duct, followed by chronic inflammation and the overexpression of mucin 5AC (MUC5AC). However, the exact mechanism responsible for the lipopolysaccharide (LPS)-induced upregulation of MUC5AC has yet to be elucidated. Specificity protein 1 (Sp1) is a ubiquitous transcription factor that plays a vital role in the regulation of a number of genes that are responsible for normal cellular function. microRNA (miR/miRNA)-130b is a member of the miRNA family. miRNAs can bind to the 3′-untralsated region (3′-UTR) of a target gene and influence its expression levels. The present study found that LPS increases the expression of MUC5AC by influencing Sp1 secretion. Chromatin immunoprecipitation-quantitative PCR experiments further verified three Sp1 binding sites in the MUC5AC promoter sequence that can regulate the expression of MUC5AC. Further analysis demonstrated that Sp1 expression was regulated by miR-130b. Luciferase experiments identified one miR-130b binding site in the Sp1 3′-UTR region. In vivo experiments also confirmed the role of the miR-130b-Sp1-MUC5AC signaling pathway in the formation of biliary stones and indicated that this pathway may provide targeted therapeutic strategies for the treatment of intrahepatic bile duct stones.