Department of Stomatology, Beijing Friendship Hospital, Capital Medical University, Beijing, China.
J Periodontal Res. 2021 Apr;56(2):256-264. doi: 10.1111/jre.12809. Epub 2021 Feb 3.
This study aimed to investigate the potential interactions among long noncoding RNA domain containing 1-antisense (lncRNA DCST1-AS1), miR-21, and periodontal ligament-associated protein-1 (PLAP-1) in periodontitis.
It has been verified that miR-21 can target PLAP-1 to regulate the osteogenic differentiation of periodontal ligament cells (PDLCs).
Differential expression of DCST1-AS1 and miR-21 in PDLCs derived from periodontitis patients and healthy controls was determined by qPCR and unpaired t test. QPCR and Western blots were conducted to evaluate the effects of overexpression of DCST1-AS1 and miR-21 on the expression of PLAP-1. CCK-8 assay was applied to evaluate the effect of DCST1-ASI, miR-21, or PLAP-1 on PDLCs' proliferation. Western blotting was conducted to detect the expression levels of CKD family (CDK4, CDK6, and CCND1).
DCST1-AS1 was downregulated in PDLCs derived from periodontitis patients, and its expression was inversely correlated with the expression of miR-2 but positively correlated with PLAP-1. Bioinformatics analysis showed that DCST1-AS1 might bind with miR-21 precursor but not mature miR-21. Transfection experiments showed that overexpression of DCST1-AS1 led to decreased expression levels of miR-21 and significantly increased the expression levels of PLAP-1 at both mRNA and protein levels, while overexpression of miR-21 resulted in a dramatic lower level of PLAP-1. CCK-8 assay indicated that overexpression of DCST1-AS1 or PLAP-1 prohibited PDLCs' proliferation. However, elevation of miR-21 had a contrary effect on the proliferation of PDLCs. And increased expression levels of DCST1-AS1 could significantly inhibit the expression of CDK4, CDK6, and CCND-1, while overexpression of miR-21 inversed the effects of DCST1-AS1.
Therefore, the expression levels of DCST1-AS1 are much lower in periodontitis patients compared to that in healthy controls, and overexpression of DCST1-AS1 can significantly elevate the expression of PLAP-1 by inhibiting miR-21 in PDLCs.
本研究旨在探讨长链非编码 RNA 结构域包含 1-反义（lncRNA DCST1-AS1）、miR-21 和牙周韧带相关蛋白-1（PLAP-1）在牙周炎中的潜在相互作用。
已经证实 miR-21 可以靶向 PLAP-1 来调节牙周韧带细胞（PDLCs）的成骨分化。
通过 qPCR 和非配对 t 检验确定来自牙周炎患者和健康对照组的 PDLCs 中 DCST1-AS1 和 miR-21 的差异表达。通过 qPCR 和 Western blot 评估过表达 DCST1-AS1 和 miR-21 对 PLAP-1 表达的影响。CCK-8 测定法用于评估 DCST1-ASI、miR-21 或 PLAP-1 对 PDLC 增殖的影响。Western blot 检测 CKD 家族（CDK4、CDK6 和 CCND1）的表达水平。
DCST1-AS1 在来自牙周炎患者的 PDLCs 中表达下调，其表达与 miR-2 的表达呈负相关，与 PLAP-1 的表达呈正相关。生物信息学分析表明，DCST1-AS1 可能与 miR-21 前体结合，但不与成熟的 miR-21 结合。转染实验表明，过表达 DCST1-AS1 导致 miR-21 的表达水平降低，并显著增加 mRNA 和蛋白水平的 PLAP-1 表达水平，而过表达 miR-21 则导致 PLAP-1 水平显著降低。CCK-8 测定表明，过表达 DCST1-AS1 或 PLAP-1 抑制 PDLCs 的增殖。然而，miR-21 的升高对 PDLCs 的增殖有相反的影响。并且 DCST1-AS1 表达水平的升高可显著抑制 CDK4、CDK6 和 CCND-1 的表达，而过表达 miR-21 则逆转了 DCST1-AS1 的作用。
因此，与健康对照组相比，牙周炎患者的 DCST1-AS1 表达水平明显较低，过表达 DCST1-AS1 通过抑制 miR-21 可显著增加 PDLCs 中 PLAP-1 的表达。
