Interference of octamer-binding transcription factor 4A and its effect on the proliferation and multiple differentiation of human dental pulp cells / 中华口腔医学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression of octamer-binding transcription factor 4A(Oct4A) in human dental pulp cells(DPC)and the effect of Oct4A on the proliferation and multiple differentiation ability of DPC.</p><p><b>METHODS</b>Expression of Oct4A in DPC was detected by realtime quantitative PCR(RT-qPCR). siRNA-Oct4A was constructed and transfected (50 nmol/L) into DPC with Lipofectamine(TM) RNAiMAX for 24, 48, 72, 96 and 120 h. The proliferation rate of DPC was examined using cell counting kit 8 (CCK-8) assay. The alizarin red staining was used to observe the formation of calcification nodules in DPC with 14 d of osteogenic induction, and oil red O staining to observe the formation of lipid droplet in DPC with 14 d of adipogenic induction. The expression of osteogenesis-related genes dentin sialophosphoprotein (DSPP) and adipogenesis-related genes lipoprotein lipase (LPL) was detected using RT-qPCR and Western blotting.</p><p><b>RESULTS</b>The expression of Oct4A reached the peak in P3 DPC (2.10 ± 0.10), which was 2.10 times as much as that in P1 (P = 0.000 vs. P1 and P7), and decreased along passages. The interference efficiency of DPC transfection peaked at 72 h (69.7%). Compared with control group and negative control group (IR-siRNA), the proliferation rate and multiple differentiation ability of DPC in interference group (Oct4A-siRNA) were downregulated (P = 0.000), and DSPP and LPL in DPC from interference group significantly decreased (P = 0.000).</p><p><b>CONCLUSIONS</b>The interference of Oct4A significantly downregulated the cell proliferation rate and multilineage differentiation capability of DPC.</p>