Abstract
BackgroundIncreasing evidence has revealed that long non-coding RNAs (lncRNAs) exert critical roles in biological mineralization. As a critical process for dentin formation, odontoblastic differentiation is regulated by complex signaling networks. The present study aimed to investigate the biological role and regulatory mechanisms of lncRNA-H19 (H19) in regulating the odontoblastic differentiation of human dental pulp stem cells (hDPSCs).MethodsWe performed lncRNA microarray assay to reveal the expression patterns of lncRNAs involved in odontoblastic differentiation. H19 was identified and verified as a critical factor by qRT-PCR. The gain- and loss-of-function studies were performed to investigate the biological role of H19 in regulating odontoblastic differentiation of hDPSCs in vitro and in vivo. Odontoblastic differentiation was evaluated through qRT-PCR, Western blot, and Alizarin Red S staining. Bioinformatics analysis identified that H19 could directly interact with miR-140-5p, which was further verified by luciferase reporter assay. After overexpression of miR-140-5p in hDPSCs, odontoblastic differentiation was determined. Moreover, the potential target genes of miR-140-5p were investigated and the biological functions of BMP-2 and FGF9 in hDPSCs were verified. Co-transfection experiments were conducted to validate miR-140-5p was involved in H19-mediated odontoblastic differentiation in hDPSCs.ResultsThe expression of H19 was significantly upregulated in hDPSCs undergoing odontoblastic differentiation. Overexpression of H19 stimulated odontoblastic differentiation in vitro and in vivo, whereas downregulation of H19 revealed the opposite effect. H19 binds directly to miR-140-5p and overexpression of miR-140-5p inhibited odontoblastic differentiation of hDPSCs. H19 acted as a miR-140-5p sponge, resulting in regulated the expression of BMP-2 and FGF9. Overexpression of H19 abrogated the inhibitory effect of miR-140-5p on odontoblastic differentiation.ConclusionOur data revealed that H19 plays a positive regulatory role in odontoblastic differentiation of hDPSCs through miR-140-5p/BMP-2/FGF9 axis, suggesting that H19 may be a stimulatory regulator of odontogenesis.
Highlights
Increasing evidence has revealed that long non-coding RNAs exert critical roles in biological mineralization
Our data revealed that H19 plays a positive regulatory role in odontoblastic differentiation of human dental pulp stem cells (hDPSCs) through miR-140-5p/bone morphogenetic protein-2 (BMP-2)/fibroblast growth factor 9 (FGF9) axis, suggesting that H19 may be a stimulatory regulator of odontogenesis
We demonstrated that H19 could accelerate odontogenic differentiation of hDPSCs via binding miR-140-5p to upregulate bone morphogenetic protein-2 (BMP-2) and fibroblast growth factor 9 (FGF9) expression
Summary
Increasing evidence has revealed that long non-coding RNAs (lncRNAs) exert critical roles in biological mineralization. As a critical process for dentin formation, odontoblastic differentiation is regulated by complex signaling networks. The present study aimed to investigate the biological role and regulatory mechanisms of lncRNA-H19 (H19) in regulating the odontoblastic differentiation of human dental pulp stem cells (hDPSCs). The complex molecular mechanisms that promote odontogenic differentiation of hDPSCs are essential for further investigation. Long non-coding RNAs (lncRNAs) play crucial roles in numerous physiological and pathological activities in cells. Recent studies have shown that lncRNAs can regulate the differentiation of stem cells [7,8,9]. Few studies have focused on the role of lncRNAs and their molecular mechanisms underlying odontoblastic differentiation of hDPSCs
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