ObjectiveThis study aims to explore the effect and molecular mechanism of long non-coding RNA (lncRNA) potassium voltage-gated channel subfamily Q member 1 overlapping transcript 1 (KCNQ1OT1) on proliferation and osteogenic differentiation in human periodontal ligament stem cells (hPDLSCs).
MethodsThe hPDLSCs of normal periodontal tissues were isolated and cultured. The mineralized solution induced the osteoblast differentiation of hPDLSCs. The down-regulation of lncRNA KCNQ1OT1, the overexpression of anti-miR-24-3p on the proliferation and the levels of osteocalcin (OCN), osteopontin (OPN) and alkaline phosphatase (ALP) of hPDLSCs were investigated. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the levels of lncRNA KCNQ1OT1, miR-24-3p, OCN, OPN, and ALP. Methyl thiazolyl tetrazolium (MTT) method was used to detect cell viability and activity. Cell proliferation was evaluated by MTT. Western blot was used to detect protein expression. The targeted relationship between lncRNA KCNQ1OT1 and miR-24-3p was detected by double-luciferase experiment.
ResultsThe expression level of lncRNA KCNQ1OT1 increased, and that of miR-24-3p decreased during the osteogenesis of hPDLSCs (P<0 .05). the down-regulation of lncrna kcnq1ot1 inhibited cell proliferation and reduced the mrna and protein expression levels of ocn, opn, and alp (P<0 .05). lncrna kcnq1ot1 targeted and regulated mir-24-3p. the overexpression of mir-24-3p inhibited cell proliferation and reduced the mrna and protein expression levels of ocn, opn, and alp (P<0 .05). inhibition of mir-24-3p reversed the effect of the down-regulation of lncrna kcnq1ot1 on cell proliferation and mrna and protein expression levels of ocn, opn, and alp (P<0 .05).
ConclusionDown-regulation of lncRNA KCNQ1OT1 inhibited the proliferation and osteogenic differentiation of hPDLSCs by targeting the up-regulated expression of miR-24-3p.
