摘要
Abstract
Objective:To discuss the effect of long non-coding RNA nuclear-enriched abundant transcript 1(Ln-cRNA NEAT1)on lipopolysaccharide(LPS)-induced osteogenic differentiation of periodontal ligament stem cells(PDLSCs)by targeting microRNA-495-3p(miR-495-3p)/Dickkopf-related protein 1(DKK1).Methods:PDLSCs were separated into Control group,LPS group,the short hairpin negative control group(sh-NC group),the short hairpin LncRNA NEAT1 lentiviral vector group(sh-NEAT1 group),sh-NEAT1+antagonist negative control group(sh-NEAT1+anti-miR-NC group),sh-NEAT1+miR-495-3p antagonist group(sh-NEAT1+anti-miR-495-3p group).Except for the Control group,all others established LPS induced cell models.qRT-PCR was performed to detect the mRNA expression of LncRNA NEAT1,miR-495-3p,and DKK1.ELISA method was performed to measure TNF-α,IL-1β,and IL-6.MTT assay and flow cytometry were performed to measure cell proliferation and apoptosis.Transwell chamber was used to detect cell migration.Alizarin red S(ARS)and alkaline phos-phatase(ALP)staining were used to detect osteogenic differentiation.Western blot was performed to detect proliferating cell nuclear antigen(PCNA),Bcl-2-associated X protein(Bax),Caspase-1,DKK1,osteocalcin(OCN),and Runt-related tran-scription factor 2(RUNX2)proteins.Dual luciferase reporter gene experiment was used to verify the targeting relationship be-tween LncRNA NEAT1,DKK1 with miR-495-3p.Results:The LncRNA NEAT1 expression was upregulated in LPS-induced PDLSCs.Silencing LncRNA NEAT1 clearly promoted LPS-induced proliferation,migration,osteogenic differentiation of PDLSCs,and inhibited inflammatory factors and cell apoptosis(P<0.05).Inhibition of miR-495-3p could reverse the effects of silencing LncRNA NEAT1 on LPS-induced proliferation,migration,osteogenic differentiation,and apoptosis of PDLSCs(P<0.05).Further analysis confirmed that LncRNA NEAT1 targeted the regulation of DKK1 through sponge like miR-495-3p(P<0.05).Conclusion:Silencing LncRNA NEAT1 can promote LPS-induced osteogenic differentiation of PDLSCs by regu-lating miR-495-3p/DKK1 axis.关键词
牙周膜干细胞/长链非编码RNA核富集转录本1/微小RNA-495-3p/Dick-kopf相关蛋白1/成骨分化Key words
Periodontal ligament stem cells/Long non-coding RNA nuclear-enriched abundant transcript 1/MicroRNA-495-3p/Dickkopf-related protein 1/Osteogenic differentiation分类
生物科学
