Liu Xiaoming, Su Kai, Kuang Shijun, Fu Min, Zhang Zhiguang
Oral Medicine Center, Oral Disease Research Institute, University of Chinese Academy of Sciences Shenzhen Hospital 4253#, Songbai Road, Matian Street, Guangming District, Shenzhen 518106, China.
Department of Mandibular Surgery, Hospital of Stomatology, Sun Yat-sen University 56#, Lingyuan West Road, Yuexiu District, Guangzhou 510055, China.
Int J Clin Exp Pathol. 2020 May 1;13(5):901-911. eCollection 2020.
Periodontitis is the second most common dental disease worldwide. TNF-α is up-regulated in periodontal disease and induces inflammation and cell apoptosis in gingival epithelial cells (GECs). miRNAs/mRNA axis play an important role in cell progression and inflammation. However, studies on the pathogenesis of periodontitisare still scarce, especially in the regulation mechanism of miRNAs.
The expression and protein level of miR-16-5p, miR-145-5p, BACH2, and caspase 3 were determined by quantitative real time PCR and western blot, respectively. Cell viability was measured by MTT assay. Cell apoptosis was detected by flow cytometry. Dual-luciferase assay was applied to verify miR-16-5p and miR-145-5p target to the 3'UTR of BACH2.
TNF-α induced miR-16-5p, miR-145-5p and caspase 3 expression, inhibited cell viability, promoted cell apoptosis in GECs. However, down-regulated miR-16-5p and miR-145-5p can restore the effects of TNF-α on GECs. In addition, dual-luciferase assay determined that BACH2 was a common target of miR-16-5p and miR-145-5p. Knockdown of BACH2 induced GECs apoptosis. Of note, cell apoptosis induced by miR-16-5p mimic, miR-145-5p mimic, and TNF-α was significantly reversed by up-regulating BACH2.
miR-16-5p and miR-145-5p mediate apoptosis induced by TNF-α in human gingival epithelial cells by targeting BACH2.
牙周炎是全球第二常见的牙科疾病。肿瘤坏死因子-α（TNF-α）在牙周疾病中上调，并诱导牙龈上皮细胞（GECs）发生炎症和细胞凋亡。微小RNA（miRNAs）/信使核糖核酸（mRNA）轴在细胞进程和炎症中发挥重要作用。然而，关于牙周炎发病机制的研究仍然匮乏，尤其是在miRNAs的调控机制方面。
分别通过定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测miR-16-5p、miR-145-5p、BACH2和半胱天冬酶3（caspase 3）的表达及蛋白水平。采用MTT法检测细胞活力。通过流式细胞术检测细胞凋亡。应用双荧光素酶报告基因检测法验证miR-16-5p和miR-145-5p对BACH2的3'非翻译区（3'UTR）具有靶向作用。
TNF-α诱导miR-16-5p、miR-145-5p和caspase 3表达，抑制GECs的细胞活力，促进细胞凋亡。然而，下调miR-16-5p和miR-145-5p可恢复TNF-α对GECs的影响。此外，双荧光素酶报告基因检测法确定BACH2是miR-16-5p和miR-145-5p的共同靶点。敲低BACH2可诱导GECs凋亡。值得注意的是，上调BACH2可显著逆转miR-16-5p模拟物、miR-145-5p模拟物和TNF-α诱导的细胞凋亡。
miR-16-5p和miR-145-5p通过靶向BACH2介导TNF-α诱导的人牙龈上皮细胞凋亡。
