miRNA-124-3p.1 Inhibits the Osteogenic and Odontogenic Differentiation of SCAPs via MACF1/smad7 Axis

Abstract

Background: Previous research has indicated that altered expression of micro-RNAs (miRNAs) is in connection with differentiation of stem cells from apical papillae (SCAPs). We investigated the mechanisms that miR-124-3p.1 inhibited osteogenic and odontogenic differentiation of SCAPs. Methods: SCAPs were isolated from dental apical papilla. MiR-124-3p.1 mimic and inhibitor were used for overexpression and knockdown assays. For overexpression and knockdown of microtubule actin cross‐linking factor 1 (MACF1), lentivirus infection and siRNA transfection were performed. Luciferase reporter assay was performed to determine the relationship between miR-124-3p.1 and MACF1. The osteogenic and odontogenic differentiation potential was analyzed by alkaline phosphatase activity analysis (ALP), alizarin red S (ARS) staining, quantitative real time reverse-transcription polymerase chain reaction (qRT-PCR), western blot and immunofluorescence (IF) staining. Results: We observed a time dependent decrease of miR‐124‐3p.1 level in mineralization induction of SCAPs. Further study found that miR‐124‐3p.1 exhibited an inhibitory effect on SCAPs osteo/odontogenic differentiation. Similarly, we found that the overexpression of miR‐124‐3p.1 dramatically inhibited MACF1 protein level in SCAPs and knockdown of miR‐124‐3p.1 significantly increased MACF1 protein level in SCAPs. Moreover, MACF1 was verified as the targeting of miR‐124‐3p.1. Meanwhile, the expression of MACF1 was related to smad7 nuclear translocation.Conclusion: Collectively, diverse data demonstrated that miR‐124‐3p.1 is a regulator of MACF1/smad7, playing plays an important role in osteogenic and odontogenic differentiation of SCAPs via MACF1/smad7 axis.