Effect of hsa-miR-378 on the Odontogenic Differentiation of Human Dental Pulp Stem Cells and Its Mechanism
MA Xiuhua, CHAO Xiaoqin, TU Zifeng, HAN Qiang, ZHOU Jie, YAO Yizhang*
This study is aimed to investigate the effect of hsa-miR-378 (human micro ribonucleic acid-378) on the differentiation of human dental pulp stem cells into odontoblasts through targeting BMP-2 (bone morphogenetic protein-2)/SMAD1 signaling pathway. Human dental pulp stem cells were isolated from dental pulp tissue and cultured. Lentivirus vectors carrying NC (negative control), hsa-miR-378 inhibitor and hsa-miR-378 mimics were transfected, and they were recorded as NC group, hsa-miR-378 inhibitor group and hsa-miR-378 mimics group, respectively. In addition, untreated cells were recorded as Blank group (only the same amount of sterile normal saline was added). The positive expression rates of STRO-1, CD105 and CD90 were high, respectively. The positive expression rates of CD34 and CD45 were low. The transfection efficiency of human dental pulp stem cells inNC group, hsa-miR-378 mimics group and hsa-miR-378 inhibitor group were high. Compared with Blank group and NC group, ALP (alkaline phosphatase) staining was lighter, and ALP activity and cell optical density were decreased (P<0.01). The number of mineralized nodules was decreased in hsa-miR-378 mimics group, and ALP staining was deepened, ALP activity and cell optical density were increased (P<0.01), and the number of mineralized nodules was increased in hsa-miR-378 inhibitor group. Compared with Blank group and NC group, the hsamiR-378 expression was increased (P<0.01), and the mRNA and protein expression levels of BMP-2, SMAD1, DMP-1 (dentin matrix protein 1), DSPP (dentin sialophosphorin) and OCN (osteocalcin) protein were decreased in hsa-miR-378 mimics group (P<0.05 or P<0.01); the hsa-miR-378 expression was decreased (P<0.01), and the mRNA and protein expression levels of BMP-2, SMAD1, DMP-1, DSPP and OCN were increased in hsa-miR-378 inhibitor group, and the differences were statistically significant (P<0.05 or P<0.01). hsa-miR-378 can mediate BMP-2 expression. Low expression of hsa-miR-378 can promote the differentiation of dental pulp stem cells into odontoblasts, which may be related to the targeted activation of BMP-2/SMAD1 signaling pathway.
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