Identification of MicroRNAs by Microarray Analysis and Prediction of Target Genes Involved in Osteogenic Differentiation of Human Periodontal Ligament Stem Cells.

BACKGROUND: The roles of microRNAs (miRNAs) in osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) remain largely unexplored. In this study, the underlying molecular mechanism of osteogenic differentiation in hPDLSCs is investigated using miRNA profiling.

METHODS: The miRNA expression profile during osteogenic differentiation was analyzed using a microarray. Target genes of miRNAs with at least two-fold change in expression (P <0.05) were predicted by bioinformatics. Six miRNAs with osteogenesis-related target genes were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR).

RESULTS: Expression of 116 miRNAs was found to be altered after osteoinduction, with 30 upregulated and 86 downregulated. Thirty-one of these miRNAs (26.7%) had osteogenesis-related target genes. Changes in expression levels of six of the 31 miRNAs (miR-654-3p, miR-4288, miR-34c-5p, miR-218-5p, miR-663a, and miR-874-3p) were validated by qRT-PCR.

CONCLUSIONS: Significant alterations in miRNA expression profiles were observed during osteogenic differentiation of hPDLSCs. These results imply that miRNAs may have regulatory effects on this process by targeting osteogenesis-related genes.