hsa-miR-93 通过 WNT/β-catenin 通路调节肝内胆管结石病中黏蛋白家族基因的表达。

hsa-miR-93 regulates MUCIN family gene expression via WNT/β-catenin pathway in intrahepatic stone disease.

机构信息

Scientific Research Experiment Center, Kunming Medical University, No. 295, Xichang Road, 65, Kunming, China.

Department of Hepatobiliary Surgery, The First Affiliated Hospital of Kunming Medical University, Kunming, China.

出版信息

Clin Res Hepatol Gastroenterol. 2018 Oct;42(5):453-461. doi: 10.1016/j.clinre.2018.03.013. Epub 2018 Apr 26.

DOI:10.1016/j.clinre.2018.03.013

PMID:29705272

Abstract

BACKGROUND

Mucin plays an essential role in the intrahepatic stone formation, but the mechanism of mucin regulation is unclear.

OBJECTIVE

To investigate the potential implication of miR-93 and WNT pathway in the regulation of intrahepatic bile duct mucin expression.

METHODS

Thirty patients with or without intrahepatic bile duct stones are involved; Reverse-transcription polymerase chain reaction was performed to evaluate the expression of MUC3, MUC4, MUC5B, MUC5AC mRNA and miR-93 levels. miR-NC or miR-93 mimics was transfected into intrahepatic biliary epithelial cells. Then mucins and Wnt pathway proteins were detected by the immunoblotting, and the target gene TCF7 were validated using the dual luciferase assay. β-catenin, wnt4, and mucins were an immunohistochemical stain of the intrahepatic biliary epithelial tissues.

RESULTS

The expression levels of MUC3, MUC4, MUC5B, and MUC5AC in patients with intrahepatic bile duct stones are higher than control, as well as Wnt pathway proteins (especially β-catenin and wnt4). Mucins levels increased in wnt4, wnt5a or SB216763-treated HIBECs, and reduced by miR-93 mimics transfection. miR-93 directly targeted TCF7 and repressed Wnt pathway protein expression, which reversed the upregulation of mucin levels induced by wnt4 or wnt5a, but not SB216763.

CONCLUSION

These results suggest a new potential mechanism in intrahepatic stones, regulating by miR-93/TCF7, non-canonical Wnt pathway, and mucins.

摘要

背景

黏蛋白在肝内结石形成中起重要作用，但黏蛋白调节的机制尚不清楚。

目的

探讨 miR-93 和 WNT 通路在调节肝内胆管黏蛋白表达中的潜在作用。

方法

纳入 30 例有或无肝内胆管结石患者，采用逆转录聚合酶链反应检测 MUC3、MUC4、MUC5B、MUC5ACmRNA 及 miR-93 水平。将 miR-NC 或 miR-93 模拟物转染至肝内胆管上皮细胞。然后通过免疫印迹检测黏蛋白和 Wnt 通路蛋白，并用双荧光素酶报告基因检测验证靶基因 TCF7。采用免疫组织化学染色检测肝内胆管上皮组织中β-catenin、wnt4 和黏蛋白。

结果

肝内胆管结石患者的 MUC3、MUC4、MUC5B 和 MUC5AC 表达水平高于对照组，Wnt 通路蛋白（特别是β-catenin 和 wnt4）也是如此。wnt4、wnt5a 或 SB216763 处理的 HIBECs 中黏蛋白水平升高，miR-93 模拟物转染后降低。miR-93 可直接靶向 TCF7 并抑制 Wnt 通路蛋白表达，逆转 wnt4 或 wnt5a 诱导的黏蛋白水平上调，但不能逆转 SB216763 的作用。