Chou Ming-Yung, Lee Chia-Hsuan, Hsieh Pei-Ling, Chao Shih-Chi, Yu Chuan-Hang, Liao Yi-Wen, Lee Shiao-Pieng, Yu Cheng-Chia, Fan Jun-Yang
School of Dentistry, Chung Shan Medical University, Taichung, Taiwan.
Department of Dentistry, Chung Shan Medical University Hospital, Taichung, Taiwan.
J Dent Sci. 2024 Oct;19(4):1999-2006. doi: 10.1016/j.jds.2024.07.002. Epub 2024 Jul 14.
BACKGROUND/PURPOSE: Oral submucous fibrosis (OSF) is a condition characterized by inflammation and excessive collagen deposition, which has been identified as a potentially malignant disorder. Recently, several microRNAs (miRNAs) have been shown to be implicated in various disorders associated with fibrosis. However, how these miRNAs modulate OSF development is poorly understood. Therefore, the study aimed to identify the specific miRNAs that contribute to the progression of OSF and to investigate their molecular mechanisms in promoting fibrosis.
The expression and clinical significance of potential pro-fibrosis miRNA in the OSF cohort and primary buccal mucosal fibroblasts were confirmed through RNA sequencing and qRT-PCR. Luciferase reporter activity assay, miRNA mimic or inhibitor, and short-hairpin RNA silencing were used to elucidate the molecular mechanism of miRNA. Transwell migration, collagen contraction, and reactive oxygen species (ROS) generation detection were used to investigate the effects of this mechanism on the myofibroblast phenotype and cellular pro-fibrosis capacity.
This study demonstrated that miR-190a was overexpressed in fibrotic buccal mucosal fibroblasts (fBMFs). Transfecting fBMFs with miR-190a inhibitor resulted in reduced cell migration, collagen gel contraction, ROS generation, and expression of fibrotic markers. Furthermore, miR-190a exerted this pro-fibrosis property by direct binding to its target, Krüppel-like factor 15 (KLF15). The results also indicated that the aberrant upregulation of miR-190a, in turn, downregulated the expression of KLF15, which resulted in the activation of myofibroblast.
Our findings demonstrated that miR-190a was involved in myofibroblast activation, suggesting that targeting the miR-190a/KLF15 axis may be a feasible approach in the therapy of OSF.
背景/目的：口腔黏膜下纤维化（OSF）是一种以炎症和胶原过度沉积为特征的疾病，已被确定为一种潜在的恶性疾病。最近，几种微小RNA（miRNA）已被证明与各种纤维化相关疾病有关。然而，这些miRNA如何调节OSF的发展尚不清楚。因此，本研究旨在确定促成OSF进展的特定miRNA，并研究它们促进纤维化的分子机制。
通过RNA测序和qRT-PCR证实了潜在促纤维化miRNA在OSF队列和原代颊黏膜成纤维细胞中的表达及其临床意义。使用荧光素酶报告基因活性测定、miRNA模拟物或抑制剂以及短发夹RNA沉默来阐明miRNA的分子机制。采用Transwell迁移、胶原收缩和活性氧（ROS）生成检测来研究该机制对肌成纤维细胞表型和细胞促纤维化能力的影响。
本研究表明，miR-190a在纤维化颊黏膜成纤维细胞（fBMFs）中过表达。用miR-190a抑制剂转染fBMFs可导致细胞迁移、胶原凝胶收缩、ROS生成减少以及纤维化标志物表达降低。此外，miR-190a通过直接与其靶标Krüppel样因子15（KLF15）结合发挥这种促纤维化特性。结果还表明，miR-190a的异常上调反过来下调了KLF15的表达，从而导致肌成纤维细胞的激活。
我们的研究结果表明，miR-190a参与了肌成纤维细胞的激活，这表明靶向miR-190a/KLF15轴可能是治疗OSF的一种可行方法。
