title: Knock-down of p300 decreases the proliferation and odontogenic differentiation potentiality of HDPCs.
abstract: OBJECTIVE
  To investigate the role of p300 in the regulation of proliferation and odontogenic differentiation of human dental pulp cells (HDPCs).
  
  
  METHODS
  The recombinant lentiviral vector pshRNA-copGFP was used to knock-down p300 expression in HDPCs. Protein level of acetylated H3 was detected. The proliferation of HDPCs was measured using the CCK8 assay. The cell cycle and apoptosis were analysed using flow cytometry and TUNEL staining, respectively. The expression levels of Cdc25A, p21(waf1) and the cleaved products of caspase 3 and caspase 7 were determined utilizing real-time quantitative polymerase chain reaction and Western blotting analysis. The alkaline phosphatase (ALP) activity was measured, and the formation of mineralized nodules was assessed using alizarin red staining after the induction of odontogenic differentiation of HDPCs. The expression levels of the odontogenic differentiation markers DMP-1, DSPP and DSP were detected utilizing real-time quantitative polymerase chain reaction and Western blotting analysis.
  
  
  RESULTS
  After p300 was knocked down in HDPCs, p300 was significantly down-regulated at both the mRNA and protein levels, and histone H3 acetylation was reduced. The proliferation capacity of HDPCs was suppressed in p300 knock-down groups. The cells were arrested in the G0/G1 phase of the cell cycle, and cell apoptosis was triggered. ALP activity, the formation of mineralized nodules and the expression levels of DMP-1, DSPP and DSP were all decreased in p300-knock-down HDPCs undergoing odontogenic differentiation.
  
  
  CONCLUSIONS
  Knocking down p300 restrains the proliferation and odontogenic differentiation potentiality of HDPCs.
authors.affiliation: Guanghua School of Stomatology & Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, China.
authors.forename: H J
authors.initials: HJ
authors.lastname: Liu
authors.affiliation: Guanghua School of Stomatology & Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, China.
authors.forename: T
authors.initials: T
authors.lastname: Wang
authors.affiliation: Guanghua School of Stomatology & Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, China.
authors.forename: Q M
authors.initials: QM
authors.lastname: Li
authors.affiliation: Guanghua School of Stomatology & Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, China.
authors.forename: X Y
authors.initials: XY
authors.lastname: Guan
authors.affiliation: Guanghua School of Stomatology & Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Xu
mesh_terms: D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000378:metabolism; D017209:Apoptosis; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D002453:Cell Cycle / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D003782:Dental Pulp / Q000166:cytology*; D050881:E1A-Associated p300 Protein / Q000378:metabolism / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D020287:In Situ Nick-End Labeling; D066298:In Vitro Techniques; D009805:Odontogenesis / Q000502:physiology*; D010750:Phosphoproteins / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000378:metabolism; D000099311:Dentin Sialophosphoprotein
chemical_list: D000469:Alkaline Phosphatase; D015415:Biomarkers; D050881:E1A-Associated p300 Protein; D016326:Extracellular Matrix Proteins; D010750:Phosphoproteins; D012795:Sialoglycoproteins; D000099311:Dentin Sialophosphoprotein; C513309:DMP1 protein, human; C494360:EP300 protein, human
keywords: apoptosis; cell cycle; odontogenic differentiation; p300; proliferation