Long Noncoding RNA X-Inactive Specific Transcript (XIST) Promotes Osteogenic Differentiation of Periodontal Ligament Stem Cells by Sponging MicroRNA-214-3p
Yimiao Feng, Pengbo Wan, Linling Yin
Abstract
BACKGROUND Osteogenic differentiation of periodontal ligament stem cells (PDLSCs) is associated with periodontitis. It has been reported that long noncoding RNA X-inactive specific transcript (lncRNA XIST) is upregulated and microRNA-214-3p (miR-214-3p) is downregulated in PDLSCs after osteogenic induction. However, whether XIST is involved in osteogenic differentiation of PDLSCs via miR-214-3p has not been reported. MATERIAL AND METHODS The protein expressions of osteogenic markers alkaline phosphatase (ALP), osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2) were examined by Western blot. The levels of miR-214-3p and XIST were determined by qRT-PCR. The relationship between miR-214-3p and XIST was evaluated by luciferase reporter, RNA immunoprecipitation, and RNA pulldown assays. RESULTS We found that XIST was increased and miR-214-3p was decreased in PDLSCs after osteogenic stimulation. Silencing of XIST decreased the protein expressions of ALP, OCN, and RUNX2, and also decreased ALP activity. Higher miR-214-3p levels also inhibited osteogenic differentiation of PDLSCs. XIST interacted with miR-214-3p and depletion of miR-214-3p mitigated XIST absence-mediated suppression of osteogenic differentiation. CONCLUSIONS XIST participates in osteogenic differentiation of PDLSCs by sponging miR-214-3p.