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长链非编码 RNA X 染色体失活特异性转录本（XIST）通过海绵吸附 microRNA-214-3p 促进牙周膜干细胞的成骨分化。

Long Noncoding RNA X-Inactive Specific Transcript (XIST) Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells by Sponging MicroRNA-214-3p.

机构信息

Department of Orthodontics, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang, China (mainland).

Department of Stomatology, Shangqiu Medical College, Shanghai, China (mainland).

出版信息

Med Sci Monit. 2020 Feb 14;26:e918932. doi: 10.12659/MSM.918932.

DOI:10.12659/MSM.918932
PMID:32057034
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7034520/
Abstract

BACKGROUND Osteogenic differentiation of periodontal ligament stem cells (PDLSCs) is associated with periodontitis. It has been reported that long noncoding RNA X-inactive specific transcript (lncRNA XIST) is upregulated and microRNA-214-3p (miR-214-3p) is downregulated in PDLSCs after osteogenic induction. However, whether XIST is involved in osteogenic differentiation of PDLSCs via miR-214-3p has not been reported. MATERIAL AND METHODS The protein expressions of osteogenic markers alkaline phosphatase (ALP), osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2) were examined by Western blot. The levels of miR-214-3p and XIST were determined by qRT-PCR. The relationship between miR-214-3p and XIST was evaluated by luciferase reporter, RNA immunoprecipitation, and RNA pulldown assays. RESULTS We found that XIST was increased and miR-214-3p was decreased in PDLSCs after osteogenic stimulation. Silencing of XIST decreased the protein expressions of ALP, OCN, and RUNX2, and also decreased ALP activity. Higher miR-214-3p levels also inhibited osteogenic differentiation of PDLSCs. XIST interacted with miR-214-3p and depletion of miR-214-3p mitigated XIST absence-mediated suppression of osteogenic differentiation. CONCLUSIONS XIST participates in osteogenic differentiation of PDLSCs by sponging miR-214-3p.

摘要

背景

牙周膜干细胞（PDLSCs）的成骨分化与牙周炎有关。据报道，在成骨诱导后，PDLSCs 中的长非编码 RNA X 失活特异性转录物（lncRNA XIST）上调，microRNA-214-3p（miR-214-3p）下调。然而，XIST 是否通过 miR-214-3p 参与 PDLSCs 的成骨分化尚未报道。

材料和方法

通过 Western blot 检测成骨标志物碱性磷酸酶（ALP）、骨钙素（OCN）和 runt 相关转录因子 2（RUNX2）的蛋白表达。通过 qRT-PCR 测定 miR-214-3p 和 XIST 的水平。通过荧光素酶报告、RNA 免疫沉淀和 RNA 下拉实验评估 miR-214-3p 和 XIST 之间的关系。

结果

我们发现，在成骨刺激后，PDLSCs 中的 XIST 增加，miR-214-3p 减少。XIST 的沉默降低了 ALP、OCN 和 RUNX2 的蛋白表达，并降低了 ALP 活性。较高的 miR-214-3p 水平也抑制了 PDLSCs 的成骨分化。XIST 与 miR-214-3p 相互作用，而 miR-214-3p 的耗竭减轻了 XIST 缺失介导的对成骨分化的抑制。

结论

XIST 通过海绵 miR-214-3p 参与 PDLSCs 的成骨分化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c57b/7034520/b6b0b5ce52cf/medscimonit-26-e918932-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c57b/7034520/98d49d1d7fb1/medscimonit-26-e918932-g001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c57b/7034520/b6b0b5ce52cf/medscimonit-26-e918932-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c57b/7034520/98d49d1d7fb1/medscimonit-26-e918932-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c57b/7034520/fb9e2529a269/medscimonit-26-e918932-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c57b/7034520/d47de181dbd9/medscimonit-26-e918932-g003.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c57b/7034520/b6b0b5ce52cf/medscimonit-26-e918932-g005.jpg

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