Objective:To investigate the effect and mechanism of long non-coding RNA(lncRNA)MEG8 on the osteogenic differentiation of hPDLSCs.Methods:hPDLSCs were isolated,purified and cultured in vitro.The expression of Osx,Runx2,Ocn,Opn and MEG8 was detected by qPCR.MEG8 overexpression and shRNA lentivirus were transfected in hPDLSCs and the transfection efficiency were identified.The RUNX2,OSX,OCN and OPN protein expression in hPDLSCs was detected by Western blot.The osteogenic differentiation of hPDLSCs was estimated by ARS(Alizarin red staining assays)demonstrated by the formation of calcified nodules.Bioinformatics methods were used to analyze candidate target genes of lncRNA MEG8,and potential target genes were further validated through dual luciferase reporter assays.The changes in osteogenic differentiation ability of hPDLSCs were analyzed after co-transfection with miR-203 mimics and lncRNA MEG8 overexpressing lentivirus.Results:The expression of Runx2,Osx,Ocn,Opn osteogenic genes and lncRNA MEG8 was significantly upregulated after osteoinduction in hPDLSCs.Knockdown of lncRNA MEG8 inhibited osteogenic differentiation of hPDLSCs and downregulated the expression of RUNX2,OSX,OCN,and OPN proteins,while overexpressing lncRNA MEG8 in hPDLSCs led to the opposite result.Through correlation analysis,it was found that miR-203 was negatively correlated with lncRNA MEG8.Further bioinformatics analysis revealed that the potential target gene of lncRNA MEG8 contained miR-203,and dual luciferase reporter experiments confirmed that miR-203 was the target gene of MEG8.The co-transfection experiment results showed that miR-203 mimics reversed the promoting effect of lncRNA MEG8 on the osteogenic differentiation of hPDLSCs.Conclusion:Our results demonstrate that lncRNA MEG8 absorbs miR-203 to promote osteogenic differentiation of hPDLSCs and shed new insights and targets for periodontitis therapeutics.
维普
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