Shen Mengdie, Meng Li'Na
Department of Gastroenterology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310006, P.R. China.
Exp Ther Med. 2019 Aug;18(2):1486-1492. doi: 10.3892/etm.2019.7707. Epub 2019 Jun 25.
The present study aimed to investigate the expression pattern and underlying mechanism of microRNA-372 (miR-372) in the progression of ulcerative colitis (UC). Reverse transcription-quantitative polymerase chain reaction was used to measure miR-372 expression levels in the blood and colonic mucosa tissue samples from patients with UC. The present study demonstrated that levels of miR-372 were significantly increased in the blood and colonic mucosa tissue samples from patients with UC compared with healthy controls. Furthermore, the level of serum miR-372 was positively correlated with the level of serum c-reactive protein. Receiver operating characteristic analysis indicated that levels of miR-372 detected in serum and tissue samples could be used to screen for patients with UC from healthy controls. These results indicated a potential role of miR-372 as a diagnostic marker and therapeutic target for patients with UC. Furthermore, a conserved miR-372 binding site in the 3'untranslated region of the NLR family pyrin domain containing 12 (NLRP12) was identified. Dual luciferase assay demonstrated that overexpression of miR-372 significantly reduced the relative luciferase activity of pmirGLO-NLRP12-3'UTR compared with control pmirGLO. In addition, western blot analysis indicated that overexpression of miR-372 significantly decreased the protein expression level of NLRP12. Therefore it was hypothesized that miR-372 may promote the progression of UC by suppressing NLRP12 protein expression and thereby inducing the excessive production of inflammatory cytokines. In conclusion, high levels of miR-372 detected in peripheral blood samples may serve a role as a potential biomarker to screen potential patients with UC from healthy controls.
本研究旨在探讨微小RNA-372(miR-372)在溃疡性结肠炎(UC)进展中的表达模式及潜在机制。采用逆转录-定量聚合酶链反应检测UC患者血液和结肠黏膜组织样本中miR-372的表达水平。本研究表明,与健康对照相比,UC患者血液和结肠黏膜组织样本中miR-372水平显著升高。此外,血清miR-372水平与血清C反应蛋白水平呈正相关。受试者工作特征分析表明,血清和组织样本中检测到的miR-372水平可用于从健康对照中筛选UC患者。这些结果表明miR-372作为UC患者诊断标志物和治疗靶点的潜在作用。此外,在含NLR家族pyrin结构域12(NLRP12)的3'非翻译区鉴定出一个保守的miR-372结合位点。双荧光素酶测定表明,与对照pmirGLO相比,miR-372过表达显著降低了pmirGLO-NLRP12-3'UTR的相对荧光素酶活性。此外,蛋白质印迹分析表明,miR-372过表达显著降低了NLRP12的蛋白表达水平。因此,推测miR-372可能通过抑制NLRP12蛋白表达从而诱导炎性细胞因子过度产生来促进UC的进展。总之,在外周血样本中检测到的高水平miR-372可能作为一种潜在生物标志物,用于从健康对照中筛选潜在的UC患者。