Hao Xinyu, Li Dongfang, Zhang Dongjiao, Jia Linglu
Department of Pediatric Dentistry, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University, Jinan, China.
Shandong Key Laboratory of Oral Tissue Regeneration, Jinan, China.
J Dent Sci. 2022 Apr;17(2):733-743. doi: 10.1016/j.jds.2021.10.014. Epub 2021 Nov 4.
BACKGROUND/PURPOSE: Dental pulp stem cells (DPSCs) are candidate seed cells for bone tissue engineering, but the molecular regulation of osteogenic differentiation in DPSCs is not fully understood. Long non-coding RNAs (lncRNAs) are important regulators of gene expression, and whether they play roles in osteogenic differentiation of DPSCs requires more study.
DPSCs were isolated and cultured. The mRNA and lncRNA expression profiles were compared through microarray assay between osteo-differentiated DPSCs and non-differentiated DPSCs. Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, Gene ontology (GO) analyses, and the mRNA-lncRNA co-expression analyses were performed for functional annotation of differentially expressed RNAs. Small interfering RNA (siRNA) was used to interfere the expression of lncRNA ENST00000533992 (also named smooth muscle-induced lncRNA or SMILR), a candidate regulator, then the osteogenic differentiation potential of DPSCs was analyzed.
DPSCs were isolated and cultured successfully. The expression of 273 mRNAs and 184 lncRNAs changed significantly in DPSCs after osteogenic induction. KEGG analyses and GO analyses showed that the differentially expressed RNAs were enriched in several pathways and biological processes. The mRNA-lncRNA co-expression network was constructed to reveal the potential relationships between mRNAs and lncRNAs. The osteogenic differentiation potential of DPSCs decreased when SMILR was interfered.
The present study provides clues for seeking for lncRNAs that participate in the regulation of osteogenic differentiation in DPSCs. LncRNA SMILR could play a role in regulating osteogenic differentiation of DPSCs.
背景/目的：牙髓干细胞（DPSCs）是骨组织工程的候选种子细胞，但DPSCs成骨分化的分子调控机制尚未完全明确。长链非编码RNA（lncRNAs）是基因表达的重要调节因子，其是否在DPSCs成骨分化中发挥作用仍需深入研究。
分离培养DPSCs。通过基因芯片分析比较成骨分化的DPSCs和未分化的DPSCs之间的mRNA和lncRNA表达谱。对差异表达的RNA进行京都基因与基因组百科全书（KEGG）分析、基因本体论（GO）分析以及mRNA-lncRNA共表达分析，以进行功能注释。使用小干扰RNA（siRNA）干扰候选调节因子lncRNA ENST00000533992（也称为平滑肌诱导lncRNA或SMILR）的表达，然后分析DPSCs的成骨分化潜能。
成功分离培养了DPSCs。成骨诱导后，DPSCs中273个mRNA和184个lncRNAs的表达发生了显著变化。KEGG分析和GO分析表明，差异表达的RNA富集于多个通路和生物学过程。构建了mRNA-lncRNA共表达网络以揭示mRNA和lncRNAs之间的潜在关系。干扰SMILR后，DPSCs的成骨分化潜能降低。
本研究为寻找参与调控DPSCs成骨分化的lncRNAs提供了线索。LncRNA SMILR可能在调控DPSCs成骨分化中发挥作用。
