Department of Stomatology, Beihua University Affiliated Hospital, Jilin 132021, China.
Anal Cell Pathol (Amst). 2021 Nov 24;2021:7890674. doi: 10.1155/2021/7890674. eCollection 2021.
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.
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-B pathway were tested by Western blot.
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-IB were elevated by ANRIL downregulation and were rescued by suppressing miR-7.
Knockdown of ANRIL inhibited osteogenic differentiation via sponging miR-7 through the NF-B pathway, suggesting that ANRIL might be a therapeutic target for periodontitis.
长链非编码 RNA（lncRNA）在牙周炎发展中失调，并参与成骨作用。本研究旨在研究 lncRNA ANRIL 在牙周韧带细胞（PDLC）中的功能及其潜在的分子机制。
首先通过 qPCR 检测 ANRIL 的水平。然后，用矿化液处理 PDLC 以诱导成骨分化。通过碱性磷酸酶（ALP）活性测定和 Western blot 检测骨形态发生蛋白 2（BMP2）、成骨转录因子 2（Osterix）和骨钙素（OCN）的蛋白水平。通过双荧光素酶报告基因实验验证 ANRIL 的靶基因。通过 qPCR 测定 miR-7 的水平，并通过 Western blot 检测 NF-B 信号通路。
在 PDL 组织中下调 ANRIL 的表达。随后，ALP 活性和 BMP2、Osterix 和 OCN 的蛋白水平增加，表明 PDLC 分化。分化的 PDLC 中 ANRIL 水平增加，而敲低则抑制成骨分化。然后，发现 miR-7 是 ANRIL 的靶基因。miR-7 在 PDL 组织中上调，在分化的 PDLC 中减少。抑制 miR-7 抑制了 ALP 活性以及 BMP2、Osterix 和 OCN 的表达。此外，抑制 miR-7 逆转了敲低 ANRIL 对成骨分化的影响。此外，下调 ANRIL 会升高 p-P65 和 p-IB 的水平，而抑制 miR-7 则可以挽救这一作用。
通过 NF-B 通路，ANRIL 通过海绵作用 miR-7 抑制成骨分化，提示 ANRIL 可能成为牙周炎的治疗靶点。
