Wu Shizhou, Liu Yunjie, Lei Lei, Zhang Hui
Department of Orthopedics, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
State Key Laboratory of Oral Diseases, Department of Preventive Dentistry, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Exp Ther Med. 2021 Jan;21(1):69. doi: 10.3892/etm.2020.9501. Epub 2020 Nov 23.
() is regarded as the major pathogen for persistent periapical periodontitis. The aim of the present study was to investigate the role of antisense RNA in the regulation of adjacent downstream genes. Reverse transcription-PCR assays were performed to validate . Adjacent downstream genes , EF1195, EF1196, and EF1197 were co-transcribed and detect antisense RNA. Northern blotting and 5'-rapid amplification of cDNA ends (5'-RACE) assays were conducted to detect and confirm a novel antisense (AS) RNA. AS overexpression mutants were constructed, and the biofilm biomass was determined using a crystal violet microtiter assay. The present study detected and confirmed a 550-bp noncoding antisense RNA with the potential to attenuate the activities of the essential response regulator . The levels of antisense RNA transcripts were inversely associated with the production of protein. It was showed that overexpression of AS leads to reduced biofilm formation and exopolysaccharide synthesis. Furthermore, the pathogenicity of was markedly decreased by AS overexpression in an periapical periodontitis model. In summary, the present study detected a novel antisense RNA that leads to a reduction in biofilm formation and the pathogenicity of . Collectively, the data suggest a role for AS as a post-transcriptional modulator of the regulator in .
（）被认为是持续性根尖周炎的主要病原体。本研究的目的是探讨反义RNA在调节相邻下游基因中的作用。进行逆转录聚合酶链反应分析以验证。相邻的下游基因EF1195、EF1196和EF1197被共同转录并检测反义RNA。进行Northern印迹和cDNA末端的5'-快速扩增（5'-RACE）分析以检测和确认一种新的反义（AS）RNA。构建AS过表达突变体，并使用结晶紫微量滴定法测定生物膜生物量。本研究检测并确认了一种550bp的非编码反义RNA，其具有减弱必需反应调节因子活性的潜力。反义RNA转录本的水平与蛋白质的产生呈负相关。结果表明，AS的过表达导致生物膜形成和胞外多糖合成减少。此外，在根尖周炎模型中，AS的过表达显著降低了（病原体）的致病性。总之，本研究检测到一种新的反义RNA，其导致生物膜形成和（病原体）致病性降低。总体而言，数据表明AS作为（病原体中）调节因子的转录后调节剂发挥作用。
