Effect of circular RNA hsa_circ_0008016 on the Osteogenic and Odontogenic Ability of Stem Cells from Apical Papilla

Abstract

This research aims to explore the role of hsa_circ_0008016 in osteogenic (OS) and odontogenic (OD) differentiation of stem cells derived from the apical papilla (SCAPs). SCAPs were isolated and cultured in either growth medium or OS and OD medium. RNA extraction was performed, followed by RNA sequencing to identify the circular RNA (circRNA) that exhibited the most significant changes between non-induced SCAPs and OS and OD-induced SCAPs, namely, hsa_circ_0008016. Subsequently, the over-expression vector pLenti-CMV-Hsa_circ_0008016-GFP-Puro was transfected into SCAPs, and the OS and OD ability of SCAPs was determined through qRT-PCR and Western blotting. In addition, the impact of hsa_circ_0008016 on 19 miRNAs were studied. Following the upregulation of hsa_circ_0008016 expression, qRT-PCR and Western blot analyses showed the expression of miRNA-660-5p, miRNA-337-3p, miRNA-376a-3p, miRNA-22-5p, bone sialoprotein (BSP) and dentin matrix protein 1 (DMP1), were increased, while the expression levels of collagenase I (COL1) and dentin sialophosphoprotein (DSPP) were decreased significantly. The target gene of miRNA-337-3p and hsa_circ_0008016 was predicted to be fibroblast growth factor receptor 1 (FGFR1). This study reveals that the hsa_circ_0008016 may enhance OS differentiation, while diminishing OD differentiation in SCAPs. This effect is likely mediated through hsa_circ_0008016 and miRNA-337-3p, potentially influencing the fibroblast growth factor (FGF)/fibroblast growth factor receptor (FGFR) signaling pathway, which warrants further validation.