title: Role of miR-424 on angiogenic potential in human dental pulp cells.
abstract: BACKGROUND
  Growing evidence shows microRNAs (miRNAs) regulate numerous cellular processes. The purpose of this study was to investigate whether miRNAs can regulate the commitment of human dental pulp cells (hDPCs) to the angiogenic fate.
  
  
  METHODS
  The hDPCs were induced to differentiate into the vascular lineage. Gene expression of endothelial markers (vWF and CD31) on day 7 after induction was analyzed by using quantitative reverse transcription-polymerase chain reaction (qRT-PCR).The miRNA expression profiling of endothelial differentiation was performed by microarray and was validated by qRT-PCR analysis. The hDPCs were infected by recombinant lentivirus to overexpress or knock down miR-424 stably, and the biological effects of miR-424 on the endothelial differentiation of hDPCs were further investigated. The tube formation ability and the amount of endothelial markers (vWF and KDR) were evaluated by Matrigel assay and Western blotting. Target genes of miR-424 were further determined by bioinformatic algorithms and Western blotting.
  
  
  RESULTS
  After endothelial differentiation, the expression of vWF and CD31 increased significantly in hDPCs. Microarray data showed that the miR-424 expression level was down-regulated on day 7. The qRT-PCR revealed a time-dependent decrease, with significant differences detected on day 1 and day 7 (P < .05). Knockdown of miR-424 expression in hDPCs promoted endothelial differentiation, with increased tube formation and up-regulated expression of vWF and KDR. In contrast, overexpression of miR-424 inhibited their differentiation. In addition, miR-424 was predicted to target vascular endothelial growth factor and KDR. Overexpression of miR-424 decreased vascular endothelial growth factor and KDR protein levels, whereas miR-424 inhibition significantly elevated them.
  
  
  CONCLUSIONS
  This study demonstrated that miR-424 may play a negative role in regulating endothelial differentiation of hDPCs, and inhibition of miR-424 may contribute to dental pulp repair and regeneration.
authors: Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology and Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China.
authors: Wei
authors: W
authors: Liu
authors: Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology and Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China.
authors: Qimei
authors: Q
authors: Gong
authors: Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology and Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China. Electronic address: lingjq@mail.sysu.edu.cn.
authors: Junqi
authors: J
authors: Ling
authors: Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology and Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China.
authors: Wen
authors: W
authors: Zhang
authors: Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology and Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China.
authors: Zhaohui
authors: Z
authors: Liu
authors: Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology and Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, Guangdong, P.R. China.
authors: Jingjing
authors: J
authors: Quan
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D002478:Cells, Cultured; D003782:Dental Pulp / Q000166:cytology*; D015536:Down-Regulation; D042783:Endothelial Cells / Q000166:cytology; D020869:Gene Expression Profiling; D055785:Gene Knockdown Techniques; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D046228:Microarray Analysis; D018919:Neovascularization, Physiologic / Q000235:genetics*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000502:physiology*; D013997:Time Factors; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000032:analysis; D014841:von Willebrand Factor / Q000032:analysis
chemical_list: C554214:MIRN424 microrna, human; D035683:MicroRNAs; D019408:Platelet Endothelial Cell Adhesion Molecule-1; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D014841:von Willebrand Factor; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: Angiogenesis; dental pulp cells; endothelial differentiation; miR-424