Interference of octamer-binding transcription factor 4A and its effect on the proliferation and multiple differentiation of human dental pulp cells
10.3760/cma.j.issn.1002-0098.2013.11.008
- VernacularTitle:干扰八聚体转录结合因子4A对人牙髓细胞增殖与多向分化能力的影响
- Author:
Li-Jing WU
1
;
Lu LIU
;
Li-Hong CHEN
;
Xi WEI
Author Information
1. 中山大学光华口腔医学院·附属口腔医院牙体牙髓科·广东省口腔医学重点实验室
- Keywords:
Cell proliferation;
Multiple differentiation;
Dental pulp cells;
Octamer-binding transcription factor 4A
- From:
Chinese Journal of Stomatology
2013;48(11):672-678
- CountryChina
- Language:Chinese
-
Abstract:
Objective 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.Methods Expression of Oct4A in DPC was detectedby realtime quantitative PCR(RT-qPCR).siRNA-Oct4A was constructed and transfected(50 nmol/L) into DPC with LipofectamineTM 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 RTqPCR and Western blotting.Results 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).Conclusions The interference of Oct4A significantly downregulated the cell proliferation rate and multilineage differentiation capability of DPC.
