Department of Cariology and Endodontology, Qingdao Stomatological Hospital Affiliated to Qingdao University, Qingdao, Shandong, China.
Department of Stomatology, Qingdao Eighth People's Hospital, Qingdao, Shandong, China.
Eur J Oral Sci. 2021 Aug;129(4):e12790. doi: 10.1111/eos.12790. Epub 2021 Jul 19.
Dental pulp stem cells (DPSCs) are multipotent and may play crucial roles in dentin-pulp regeneration. Recent studies have revealed that long noncoding RNAs (lncRNAs) are implicated in the osteogenic differentiation of DPSCs. However, the specific role and potential mechanisms of the lncRNA trihydroxyacetophenone domain containing nine antisense RNA 1 (THAP9-AS1) during osteogenic differentiation of DPSCs remain unknown. In the present study, we determined that THAP9-AS1 expression was upregulated during osteogenic differentiation of DPSCs. Moreover, we investigated the biological functions of THAP9-AS1 during osteogenic differentiation of DPSCs by loss-of-function assays. THAP9-AS1 knockdown inhibited osteogenic differentiation of DPSCs by decreasing alkaline phosphatase activity, alkaline phosphatase-positive cell ratio, mineralizing matrix and mRNA, and protein levels of early osteogenic-markers. We also found that THAP9-AS1 interacted with miR-652-3p, whose downstream gene target is vascular endothelial growth factor A (VEGFA). In addition, rescue assays indicated that VEGFA rescued the effects of THAP9-AS1 knockdown during osteogenic differentiation of DPSCs. In summary, we verified that knockdown of THAP9-AS1 inhibits osteogenic differentiation of DPSCs via the miR-652-3p/VEGFA axis. Our findings may be helpful to extend research on the mechanisms underlying osteogenic differentiation of DPSCs.
牙髓干细胞（DPSCs）具有多能性，可能在牙本质-牙髓再生中发挥关键作用。最近的研究表明，长非编码 RNA（lncRNA）参与 DPSCs 的成骨分化。然而，lncRNA 三羟基苯乙酮结构域包含九个反义 RNA 1（THAP9-AS1）在 DPSCs 成骨分化中的具体作用和潜在机制尚不清楚。在本研究中，我们确定 THAP9-AS1 的表达在 DPSCs 的成骨分化过程中上调。此外，我们通过功能丧失试验研究了 THAP9-AS1 在 DPSCs 成骨分化过程中的生物学功能。THAP9-AS1 敲低通过降低碱性磷酸酶活性、碱性磷酸酶阳性细胞比例、矿化基质和早期成骨标志物的 mRNA 和蛋白水平来抑制 DPSCs 的成骨分化。我们还发现 THAP9-AS1 与 miR-652-3p 相互作用，miR-652-3p 的下游基因靶标是血管内皮生长因子 A（VEGFA）。此外，挽救试验表明，VEGFA 挽救了 THAP9-AS1 敲低对 DPSCs 成骨分化的影响。综上所述，我们验证了敲低 THAP9-AS1 通过 miR-652-3p/VEGFA 轴抑制 DPSCs 的成骨分化。我们的研究结果可能有助于扩展对 DPSCs 成骨分化机制的研究。
