title: [Expression profiles and bioinformatic analysis of miRNA in human dental pulp cells during endothelial differentiation].
abstract: OBJECTIVE
  To investigate the differential expression profile and bioinformatic analysis of microRNA (miRNA) in human dental pulp cells (DPC) during endothelial differentiation.
  
  
  METHODS
  DPC were cultured in endothelial induction medium (50 µg/L vascular endothelial growth factor, 10 µg/L basic fibroblast growth factor and 2% fetal calf serum) for 7 days. Meanwhile non-induced DPC were used as control.Quantitative real-time PCR (qRT-PCR) was applied to detect vascular endothelial marker genes [CD31, von Willebrand factor (vWF) and vascular endothelial-cadherin (VE-cadherin)] and in vitro tube formation on matrigel was used to analyze the angiogenic ability of differentiated cells. And then miRNA expression profiles of DPC were examined using miRNA microarray and then the differentially expressed miRNA were validated by qRT-PCR. Furthermore, bioinformatic analysis was employed to predict the target genes of miRNA and to analyze the possible biological functions and signaling pathways that were involved in DPC after induction.
  
  
  RESULTS
  The relative mRNA level of CD31, vWF and VE-cadherin in the control group were (3.48 ± 0.22) ×10(-4), (3.13 ± 0.31) ×10(-4) and (39.60 ± 2.36) ×10(-4), and (19.57 ± 2.20) ×10(-4), (48.13 ± 0.54) ×10(-4) and (228.00 ± 8.89) ×10(-4) in the induced group. The expressions of CD31, vWF and VE-cadherin were increased significantly in endothelial induced DPC compared to the control group (P < 0.05). For in vitro tube formation assay, tubular structures were formed on the matrigel by differentiated DPC. A total of 47 miRNA were differentially expressed, in which 15 miRNA were up-regulated and 32 miRNAs down-regulated in differentiated DPC compared with the control. Of these, 4 miRNA were confirmed by qRT-PCR. The target genes of differential miRNA were predicted to associate with several biological functions, such as the regulation of transcription, cell motion, blood vessel morphogenesis, angiogenesis and cytoskeletal protein, and signaling pathways including the mitogen-activated protein kinase (MAPK) and the Wnt signaling pathway.
  
  
  CONCLUSIONS
  The differential miRNA expression identified in this study may be involved in governing DPC endothelial differentiation, thus contributing to the future research on regulatory mechanisms in dental pulp angiogenesis.
authors.affiliation: Department of Operative Dentistry and Endodontics,Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University & Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China.
authors.forename: Qimei
authors.initials: Q
authors.lastname: Gong
authors.affiliation: Department of Operative Dentistry and Endodontics,Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University & Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Department of Operative Dentistry and Endodontics,Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University & Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China.
authors.forename: Jinming
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Operative Dentistry and Endodontics,Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University & Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China. Email: lingjq@mail.sysu.edu.cn.
authors.forename: Junqi
authors.initials: J
authors.lastname: Ling
mesh_terms: D015703:Antigens, CD; D015820:Cadherins; D002454:Cell Differentiation; D003094:Collagen; D019295:Computational Biology; D003782:Dental Pulp / Q000378:metabolism*; D004338:Drug Combinations; D016222:Fibroblast Growth Factor 2; D006801:Humans; D007797:Laminin; D035683:MicroRNAs; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000096:biosynthesis; D011509:Proteoglycans; D012333:RNA, Messenger; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction; D042461:Vascular Endothelial Growth Factor A; D060449:Wnt Signaling Pathway; D014841:von Willebrand Factor; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D003094:Collagen; D004338:Drug Combinations; D016222:Fibroblast Growth Factor 2; D007797:Laminin; D035683:MicroRNAs; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011509:Proteoglycans; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D014841:von Willebrand Factor; D000099152:Cadherin 5; C059485:matrigel