Illuminating the mechanisms of odontoblast differentiation of human dental pulp stem cells (hDPSCs) is the key to find therapeutic clues to promote odontogenesis. LncRNAs play a regulatory role in odontoblast differentiation. Here, we identified a novel lncRNA, named lncRNA CALB2. It was up-regulated in odontoblast-differentiated hDPSCs and potentially interacted with miR-30b-3p and RUNX2. Via gain- and loss-of-function approaches, we found lncRNA CALB2 significantly promoted the odontoblast differentiation of hDPSCs. Then, dual luciferase reporter assay and RNA immunoprecipitation assay revealed that both lncRNA CALB2 and RUNX2 mRNA could directly bind to miR-30b-3p via the same binding sites. Interestingly, miR-30b-3p in hDPSCs was down-regulated and RUNX2 was up-regulated during odontoblast differentiation. Moreover, lncRNA CALB2 knockdown significantly reduced the protein level of RUNX2, DSPP and DMP-1, while miR-30b-3p inhibitor rescued the reduction. Furthermore, miR-30b-3p exerted an inhibitory effect on odontoblast differentiation, which could be reversed by lncRNA CALB2. Collectively, these findings indicate that the newly identified lncRNA CALB2 acts as a miR-30b-3p sponge to regulate RUNX2 expression, thus promoting the odontoblast differentiation of hDPSCs. LncRNA CALB2/miR-30b-3p/RUNX2 axis could be a novel therapeutic target for accelerating odontogenesis.

阐明人牙髓干细胞（hDPSCs）成牙本质细胞分化的机制是寻找促进牙形成的治疗线索的关键。LncRNA 在成牙本质细胞分化中起调节作用。在这里，我们鉴定了一种新的 lncRNA，命名为 lncRNA CALB2。它在成牙本质细胞分化的 hDPSC 中上调，并可能与 miR-30b-3p 和RUNX2相互作用。通过功能获得和功能丧失的方法，我们发现 lncRNA CALB2 显着促进了 hDPSCs 的成牙本质细胞分化。然后，双荧光素酶报告基因测定和 RNA 免疫沉淀测定显示 lncRNA CALB2 和RUNX2 mRNA 都可以通过相同的结合位点直接结合 miR-30b-3p。有趣的是，hDPSCs 中的 miR-30b-3p 被下调，而RUNX2在成牙本质细胞分化过程中被上调。此外，lncRNA CALB2 敲低显着降低了 RUNX2、DSPP 和 DMP-1 的蛋白质水平，而 miR-30b-3p 抑制剂则挽救了这种降低。此外，miR-30b-3p对成牙本质细胞分化有抑制作用，lncRNA CALB2可以逆转这种作用。总的来说，这些发现表明新发现的 lncRNA CALB2 充当 miR-30b-3p 海绵来调节 RUNX2 表达，从而促进 hDPSCs 的成牙本质细胞分化。LncRNA CALB2/miR-30b-3p/RUNX2 轴可能是加速牙形成的新治疗靶点。