Junli Yue1, Puyu Wang1, Qingchun Hong1, Qian Liao1, Li Yan1, Weizhe Xu1, Xi Chen1, Qinghua Zheng1, Lan Zhang2, Dingming Huang3. 1. State Key Laboratory of Oral Diseases, Department of Endodontics, West China Hospital of Stomatology, Sichuan University, Sichuan, China. 2. State Key Laboratory of Oral Diseases, Department of Endodontics, West China Hospital of Stomatology, Sichuan University, Sichuan, China. Electronic address: zlnancy914@sina.com. 3. State Key Laboratory of Oral Diseases, Department of Endodontics, West China Hospital of Stomatology, Sichuan University, Sichuan, China. Electronic address: dingminghuang@163.com.
Abstract
INTRODUCTION: MicroRNA-335-5p has been reported to regulate osteogenic and chondrogenic differentiations of mesenchymal stem cells. The aim of this study was to explore the function and regulation mechanism of miR-335-5p in apical periodontitis (AP). METHODS: Total RNAs were extracted from human periodontal ligament fibroblasts (HPDLFs), 10 AP tissues, and 6 healthy periodontal ligament tissues using lysis buffer. Gene expression was detected using real-time polymerase chain reaction. The Dual Luciferase Assay (Promega, Madison, WI) was used to test miR-335-5p directly targeted urokinase-type plasminogen activator receptor (uPAR) and the receptor activator of nuclear factor kappa-B ligand (RANKL). Western Blot was used to detect protein expressions of RANKL, uPAR, and the fragile X-related 1 gene (FXR1). The enzyme-linked immunosorbent assay was used to detect the secretions of interleukin 6, tumor necrosis factor alpha, and RANKL. Data were analyzed using the Student t test. RESULTS: miR-335-5p acted as a positive mediator in HPDLF inflammation (P < .05). Two targets of miR-335-5p, uPAR and RANKL, were identified. Interestingly, uPAR was repressed by miR-335-5p at the basal level, but it can be relieved from miR-335-5p-mediated repression, which is called derepression, when HPDLFs were subjected to lipopolysaccharide stimulation. miR-335-5p promoted RANKL in HPDLFs regardless of whether or not it was under inflammatory conditions (P < .05). We proved FXR1 was responsible for the derepression of uPAR from miR-335-5p (P < .01). Both FXR1 and uPAR were positive mediators in HPDLF inflammation (P < .05). miR-335-5p, uPAR, RANKL, and FXR1 had the same expression profiles in HPDLF inflammation and AP tissues (P < .05). CONCLUSIONS: Our data showed that miR-335-5p may play dual roles in AP, and it might be considered as a target for therapeutic potency in clinical applications.
INTRODUCTION: MicroRNA-335-5p has been reported to regulate osteogenic and chondrogenic differentiations of mesenchymal stem cells. The aim of this study was to explore the function and regulation mechanism of miR-335-5p in apical periodontitis (AP). METHODS: Total RNAs were extracted from human periodontal ligament fibroblasts (HPDLFs), 10 AP tissues, and 6 healthy periodontal ligament tissues using lysis buffer. Gene expression was detected using real-time polymerase chain reaction. The Dual Luciferase Assay (Promega, Madison, WI) was used to test miR-335-5p directly targeted urokinase-type plasminogen activator receptor (uPAR) and the receptor activator of nuclear factor kappa-B ligand (RANKL). Western Blot was used to detect protein expressions of RANKL, uPAR, and the fragile X-related 1 gene (FXR1). The enzyme-linked immunosorbent assay was used to detect the secretions of interleukin 6, tumor necrosis factor alpha, and RANKL. Data were analyzed using the Student t test. RESULTS:miR-335-5p acted as a positive mediator in HPDLF inflammation (P < .05). Two targets of miR-335-5p, uPAR and RANKL, were identified. Interestingly, uPAR was repressed by miR-335-5p at the basal level, but it can be relieved from miR-335-5p-mediated repression, which is called derepression, when HPDLFs were subjected to lipopolysaccharide stimulation. miR-335-5p promoted RANKL in HPDLFs regardless of whether or not it was under inflammatory conditions (P < .05). We proved FXR1 was responsible for the derepression of uPAR from miR-335-5p (P < .01). Both FXR1 and uPAR were positive mediators in HPDLF inflammation (P < .05). miR-335-5p, uPAR, RANKL, and FXR1 had the same expression profiles in HPDLF inflammation and AP tissues (P < .05). CONCLUSIONS: Our data showed that miR-335-5p may play dual roles in AP, and it might be considered as a target for therapeutic potency in clinical applications.
Authors: Jennie Ong; Anke van den Berg; Alen Faiz; Ilse M Boudewijn; Wim Timens; Cornelis J Vermeulen; Brian G Oliver; Klaas Kok; Martijn M Terpstra; Maarten van den Berge; Corry-Anke Brandsma; Joost Kluiver Journal: Int J Mol Sci Date: 2019-10-18 Impact factor: 5.923
Authors: Tadkamol Krongbaramee; Min Zhu; Qingwen Qian; Zeyuan Zhang; Steven Eliason; Yi Shu; Fang Qian; Adil Akkouch; Dan Su; Brad A Amendt; Ling Yang; Liu Hong Journal: Mol Ther Nucleic Acids Date: 2021-02-04 Impact factor: 8.886