Downregulation of microRNA‐143‐5p is required for the promotion of odontoblasts differentiation of human dental pulp stem cells through the activation of the mitogen‐activated protein kinases 14‐dependent p38 mitogen‐activated protein kinases signaling pathway

Abstract

MicroRNAs (miRNAs) play critical roles in various biological processes including cell differentiation. Some researchers suggested that the p38 mitogen-activated protein kinases (MAPK) signaling pathway had an effect on regulating the odontoblastic differentiation of human dental pulp stem cells (hDPSCs). This study focuses on the effects of miR-143-5p on hDPSCs by regulating the p38 MAPK signaling pathway. The targeting relationship of MAPK14 and miR-143-5p targets were verified by TargetScan and dual-luciferase reporter gene assay. Through overexpression of miR-143-5p or silencing of miR-143-5p, expressions of miR-143-5p, MAPK14, Ras, MAPK kinase (MKK) 3/6, dentin sialophosphoprotein (DSPP), alkaline phosphatase (ALP), and osteocalcin (OCN) were detected by reverse transcription quantitative polymerase chain reaction. Protein expressions of MAPK14, Ras, and MKK3/6 were determined by western blot analysis. ALP and alizarin red S staining were used to detect mineralization. Initially, MAPK14 was found to be negatively regulated by miR-143-5p. Meanwhile, the upregulated miR-143-5p decreased the p38 MAPK signaling pathway related genes (MAPK14, Ras, and MKK3/6) and odontoblastic differentiation markers (ALP, DSPP, and OCN) expression. On the contrary, the downregulated miR-143-5p increased the p38 MAPK signaling pathway related genes (MAPK14, Ras, and MKK3/6) and odontoblastic differentiation markers (ALP, DSPP, and OCN) expression. Furthermore, ALP activity and mineralized nodules increased after downregulation of miR-143-5p, and after its upregulation, ALP activity and mineralized nodules decreased. Our data suggest that poor expression of miR-143-5p promotes hDPSCs odontoblastic differentiation through the activation of the p38 MAPK signaling pathway by upregulating MAPK14. K E Y W O R D S human dental pulp stem cells (hDPSCs), mitogen-activated protein kinases (MAPK) 14, microRNA-143-5p (miR-143-5p), odontoblastic differentiation, p38 MAPK signaling pathway J Cell Physiol. 2019;234:4840-4850. wileyonlinelibrary.com/journal/jcp 4840 |

Epigenetic Regulation of Dental Pulp Stem Cell Fate

“…It has been reported that miR-143-5p binds to MAPK14 and reduces its expression. Thus, miR-143-5p knockdown increases MAPK14 expression and activates the p38 MAPK signaling pathway, consequently enhancing the odontogenic differentiation [ 151 ]. In addition, by directly targeting tumor necrosis factor- α (TNF- α ), miR-143 blockades the NF- κ B signaling pathway and suppresses the osteogenic differentiation [ 152 ].…”

Noncoding RNAs: new insights into the odontogenic differentiation of dental tissue-derived mesenchymal stem cells

“…This suggests that miR-143-5p could be developed as a target of genetically modified stem cell therapy for pulp regeneration. Another study of Wang et al [55] also identified the inhibitory role of miR-143-5p in the odontogenic differentiation of human DPSCs. They showed that the downregulated miR-143-5p expression induced the expression of the p38 MAPK signaling pathway-related gene MAPK14 and odontogenesis-related markers.…”

Genome-wide identification of long noncoding RNAs and their competing endogenous RNA networks involved in the odontogenic differentiation of human dental pulp stem cells

“…In the research on ncRNAs and their roles in the odontogenic differentiation of hDPSCs, the most studied are miRNAs. miR-223-3p and miR-21 have been shown to promote odontogenic differentiation, while miR-143-5p, miR-140-5p, and miR-488 inhibit this process [41][42][43][44][45][46]. In 2016, Chen et al found that lncRNA differentiation antagonizing non-protein coding RNA (DANCR) inhibited the odontogenic differentiation of human dental pulp cells by inactivating the Wnt/β-catenin signaling pathway [16].…”