title: Down-regulated non-coding RNA (lncRNA-ANCR) promotes osteogenic differentiation of periodontal ligament stem cells.
abstract: OBJECTIVE
  Our studies aimed to figure out how anti-differentiation noncoding RNA (ANCR) regulates the proliferation and osteogenic differentiation of periodontal ligament stem cells (PDLSCs).
  
  
  METHODS
  In this study, we used lentivirus infection to down-regulate the expression of ANCR in PDLSCs. Then we compared the proliferation of control cells and PDLSC/ANCR-RNAi cells by Cell Counting Kit-8. And the osteogenic differentiation of control cells and PDLSC/ANCR-RNAi cells were evaluated by Alkaline phosphatase (ALP) activity quantification and Alizarin red staining. WNT inhibitor was used to analyze the relationship between ANCR and canonical WNT signalling pathway. The expression of osteogenic differentiation marker mRNAs, DKK1, GSK3-β and β-catenin were evaluated by qRT-PCR.
  
  
  RESULTS
  The results showed that down-regulated ANCR promoted proliferation of PDLSCs. Down-regulated ANCR also promoted osteogenic differentiation of PDLSCs by up-regulating osteogenic differentiation marker genes. After the inhibition of canonical WNT signalling pathway, the osteogenic differentiation of PDLSC/ANCR-RNAi cells was inhibited too. qRT-PCR results also demonstrated that canonical WNT signalling pathway was activated for ANCR-RNAi on PDLSCs during the procedure of proliferation and osteogenic induction.
  
  
  CONCLUSIONS
  These results indicated that ANCR was a key regulator of the proliferation and osteogenic differentiation of PDLSCs, and its regulating effects was associated with the canonical WNT signalling pathway, thus offering a new target for oral stem cell differentiation studies that could also facilitate oral tissue engineering.
authors.affiliation: State Key Laboratory of Military Stomatology, Department of Operative Dentistry and Endodontics, School of Stomatology, 4th Military Medical University, 145 Changle West Road, Xi'an 710032, Shannxi, PR China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Jia
authors.affiliation: State Key Laboratory of Military Stomatology, Department of Operative Dentistry and Endodontics, School of Stomatology, 4th Military Medical University, 145 Changle West Road, Xi'an 710032, Shannxi, PR China.
authors.forename: Wenkai
authors.initials: W
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Military Stomatology, Department of Operative Dentistry and Endodontics, School of Stomatology, 4th Military Medical University, 145 Changle West Road, Xi'an 710032, Shannxi, PR China. Electronic address: Longxing_Ni@hotmail.com.
authors.forename: Longxing
authors.initials: L
authors.lastname: Ni
mesh_terms: D000328:Adult; D000469:Alkaline Phosphatase / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D015536:Down-Regulation; D006801:Humans; D016130:Immunophenotyping; D010012:Osteogenesis / Q000502:physiology*; D010513:Periodontal Ligament / Q000166:cytology*; D062085:RNA, Long Noncoding / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction; D013234:Stem Cells / Q000502:physiology*; D060449:Wnt Signaling Pathway / Q000502:physiology*
chemical_list: D062085:RNA, Long Noncoding; D000469:Alkaline Phosphatase
keywords: Canonical WNT signalling pathway; LncRNA; Osteogenesis; Periodontal ligament stem cells