The State Key Laboratory Breeding Base of Basic Science of Stomatology and Key Laboratory of Oral Biomedicine of Ministry of Education, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan, 430079, China.
Biochem Biophys Res Commun. 2021 Mar 19;545:105-111. doi: 10.1016/j.bbrc.2021.01.076. Epub 2021 Feb 3.
Transforming growth factor β1 (TGF-β1) is one of the broad-spectrum growth-promoting factors that participate in tooth development. The influence of TGF-β1 on the odontoblastic differentiation is still controvercy. Mouse primary dental papilla cells (mDPCs) as well as an immortalized mouse dental papilla cell line (mDPC6Ts) were treated with exogenous TGF-β1 during odontoblastic differentiation. RT-qPCR, Western blot, alizarin red staining and ALP staining were carried out to investigate the influence of TGF-β1 on odontoblastic differentiation. IPO7, important for SMAD complex translocation was also detected in mDPCs and mDPC6Ts in response to TGF-β1. After silencing IPO7 by transfection, the translocation process of P-SMAD2 was investigated by nuclear and cytoplasmic extraction as well as co-immunoprecipitation assay. The odontogenic markers, mineralization and IPO7 expression were significantly up-regulated in TGF-β1-treated mDPCs while down-regulated in mDPC6Ts. The total level of P-SMAD2 was not influenced by IPO7 in mDPCs, however, IPO7 could bind to P-SMAD2 and affect the nuclear-cytoplasm-shuttling of P-SMAD2. Our data demonstrated that TGF-β1 plays opposite roles in odontoblast differentiation in mDPCs and immortalized mouse dental papilla cell line (mDPC6Ts), which is determined by IPO7.
转化生长因子 β1（TGF-β1）是参与牙齿发育的广谱促生长因子之一。TGF-β1 对成牙本质细胞分化的影响仍存在争议。在成牙本质细胞分化过程中，用外源性 TGF-β1 处理小鼠原代牙髓细胞（mDPCs）和永生化小鼠牙髓细胞系（mDPC6Ts）。进行 RT-qPCR、Western blot、茜素红染色和 ALP 染色，以研究 TGF-β1 对成牙本质细胞分化的影响。还检测了 IPO7，其对于 SMAD 复合物易位很重要，在 mDPCs 和 mDPC6Ts 中对 TGF-β1 也有反应。通过转染沉默 IPO7 后，通过核质提取和共免疫沉淀实验研究 P-SMAD2 的易位过程。TGF-β1 处理的 mDPCs 中成牙本质标志物、矿化和 IPO7 的表达明显上调，而 mDPC6Ts 中则下调。在 mDPCs 中，IPO7 对 P-SMAD2 的总水平没有影响，但是，IPO7 可以与 P-SMAD2 结合并影响 P-SMAD2 的核质穿梭。我们的数据表明，TGF-β1 在 mDPCs 和永生化小鼠牙髓细胞系（mDPC6Ts）中成牙本质细胞分化中发挥相反的作用，这取决于 IPO7。
