Abstract
Background Long noncoding RNAs (lncRNAs) are dysregulated in periodontitis development and involved in osteogenesis. The current study was aimed at investigating the function of lncRNA ANRIL in periodontal ligament cells (PDLCs) and potential molecular mechanisms. Methods Firstly, the level of ANRIL was tested by qPCR. Then, PDLCs were treated with a mineralizing solution to induce osteogenic differentiation. ALP activity was measured, and protein levels of BMP2, Osterix, and OCN were measured by Western blot. A target of ANRIL was verified using dual-luciferase reporter assay. miR-7 level was measured by qPCR, and the signals of the NF-κB pathway were tested by Western blot. Results ANRIL expression was downregulated in PDL tissues. Next, ALP activity and protein levels of BMP2, Osterix, and OCN were increased to show that PDLCs were differentiated. ANRIL level was increased in differential PDLCs, in which knockdown inhibited osteogenic differentiation. Then, miR-7 was found as a target of ANRIL. The miR-7 level was upregulated in PDL tissues and reduced in differential PDLCs. Inhibition of miR-7 suppressed ALP activity and BMP2, Osterix, and OCN expression. Moreover, inhibition of miR-7 reversed the effects on the osteogenic differentiation induced by knockdown of ANRIL. Besides, the levels of p-P65 and p-IκBα were elevated by ANRIL downregulation and were rescued by suppressing miR-7. Conclusions Knockdown of ANRIL inhibited osteogenic differentiation via sponging miR-7 through the NF-κB pathway, suggesting that ANRIL might be a therapeutic target for periodontitis.
Highlights
Long noncoding RNAs are dysregulated in periodontitis development and involved in osteogenesis
According to the results of Quantitative polymerase chain reaction WT (qPCR), the antisense noncoding RNA in the INK4 locus (ANRIL) level was reduced in periodontal ligament (PDL) tissues of patients with periodontitis, compared with the healthy control group (Figure 1)
To investigate the osteogenic differentiation capability, periodontal ligament cells (PDLCs) derived from PDL tissues were cultured in DMEM with β-glycerophosphate, vitamin C, and dexamethasone
Summary
Long noncoding RNAs (lncRNAs) are dysregulated in periodontitis development and involved in osteogenesis. ALP activity and protein levels of BMP2, Osterix, and OCN were increased to show that PDLCs were differentiated. ANRIL level was increased in differential PDLCs, in which knockdown inhibited osteogenic differentiation. The miR-7 level was upregulated in PDL tissues and reduced in differential PDLCs. Inhibition of miR-7 suppressed ALP activity and BMP2, Osterix, and OCN expression. Inhibition of miR-7 reversed the effects on the osteogenic differentiation induced by knockdown of ANRIL. Knockdown of ANRIL inhibited osteogenic differentiation via sponging miR-7 through the NF-κB pathway, suggesting that ANRIL might be a therapeutic target for periodontitis. Periodontitis is a chronic, nonspecific, and multifactorial inflammatory disease associated with periodontal support tissue It will cause pathological loss of the periodontal ligament and alveolar bone, leading to teeth loss [1]. The treatment of periodontitis is complex, and there is still a lack of early screening biomarkers and therapeutic targets
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