Xie Bing, Yang Zhen-Yu, Tang Xu-Na
Department of Dental and Endodontic Diseases, Nanjing Stomatological Hospital, Medical School of Nanjing University. Nanjing 210008, Jiangsu Province, China. E-mail:
Shanghai Kou Qiang Yi Xue. 2023 Apr;32(2):147-153.
To investigate the possible role of circRASA2 in periodontitis and its potential regulatory mechanism.
Periodontitis cell model was established by lipopolysaccharide(LPS)-induced periodontal ligament cells(PDLCs). Cell proliferation activity was detected by CCK-8 assay, cell migration ability was detected by Transwell chamber assay, and the expression of osteogenic differentiation-related proteins in cells was detected by Western blot. The target miRNA of circRASA2 and its downstream target genes were predicted using the databases circinteractome and starBase, respectively, and the targeting relationship between the target genes was verified by dual-luciferase reporter gene experiment. GraphPad Prism 8.0 software package was used to analyze the data.
circRASA2 was highly expressed in LPS-treated PDLCs cells. LPS-induced PDLCs cell proliferation activity, migration ability and osteogenic differentiation ability decreased, while knockdown of circRASA2 promoted proliferation, migration and osteogenic differentiation ability of PDLCs under LPS treatment. circRASA2 targeted and negatively regulated the expression of miR-543, and overexpression of miR-543 promoted proliferation, migration and osteogenic differentiation of PDLCs under LPS treatment. TRAF6 was a downstream target gene of miR-543, knockdown of circRASA2 down-regulated the expression of TRAF6 through the sponge action of miR-543. Overexpression of TRAF6 reversed the promotion of circRASA2 knockdown on proliferation, migration and osteogenic differentiation of PDLCs.
circRASA2 accelerated the pathological process of periodontitis in vitro through miR-543/TRAF6 axis, and might improve periodontitis by targeting down the expression of circRASA2.
探讨环状RASA2（circRASA2）在牙周炎中的可能作用及其潜在调控机制。
通过脂多糖（LPS）诱导牙周膜细胞（PDLCs）建立牙周炎细胞模型。采用CCK-8法检测细胞增殖活性，Transwell小室法检测细胞迁移能力，蛋白质免疫印迹法检测细胞中成骨分化相关蛋白的表达。分别使用circinteractome和starBase数据库预测circRASA2的靶标微小RNA（miRNA）及其下游靶基因，并通过双荧光素酶报告基因实验验证靶基因之间的靶向关系。采用GraphPad Prism 8.0软件包进行数据分析。
circRASA2在LPS处理的PDLCs细胞中高表达。LPS诱导的PDLCs细胞增殖活性、迁移能力和成骨分化能力降低，而敲低circRASA2可促进LPS处理下PDLCs的增殖、迁移和成骨分化能力。circRASA2靶向并负调控miR-543的表达，miR-543过表达可促进LPS处理下PDLCs的增殖、迁移和成骨分化。肿瘤坏死因子受体相关因子6（TRAF6）是miR-543的下游靶基因，敲低circRASA2通过miR-543的海绵作用下调TRAF6的表达。TRAF6过表达逆转了circRASA2敲低对PDLCs增殖、迁移和成骨分化的促进作用。
circRASA2通过miR-543/TRAF6轴在体外加速牙周炎的病理进程，下调circRASA2的表达可能改善牙周炎。
