title: Antidifferentiation Noncoding RNA Regulates the Proliferation of Osteosarcoma Cells.
abstract: BACKGROUND
  Antidifferentiation noncoding RNA (ANCR), a newly identified long noncoding RNA (lncRNA), plays a critical role for stem cells to maintain undifferentiated cell state. However, the functions of ANCR in human cancers have not been reported. This study is designed to explore the role of ANCR in osteosarcoma.
  
  
  METHODS
  Lentivirus-mediated (shRNA) was applied to silence ANCR in the human osteosarcoma cell lines U2OS and Saos-2. Cell viability was measured by MTT assay. Colony-forming ability was measured by colony formation assay. Cell cycle progression was determined by flow cytometry with propidium iodide staining. In addition, cell cycle makers, including p21, CDK2, and CDK4, were investigated in ANCR silencing U2OS cells by real time PCR (RT-PCR) analysis.
  
  
  RESULTS
  In this study, we first proved that lentivirus-mediated shRNA specifically suppressed the expression level of ANCR in U2OS and Saos-2 cells. Further investigations revealed that knockdown of ANCR significantly inhibited the proliferation of U2OS and Saos cells and colony formation of U2OS cells. Moreover, the cell cycle of U2OS cells was arrested at G0/G1 phase after ANCR knockdown. Furthermore, the expression level of p21 was increased and CDK2 was decreased in ANCR knock-down cells.
  
  
  CONCLUSIONS
  Our data indicated that ANCR might be an oncogenic lncRNA that promoted proliferation of osteosarcoma. The potential application of ANCR-targeted therapy using the lentivirus-mediated shRNA approach is worth further investigations in preclinical and clinical studies.
authors.affiliation: 1 Department of Orthopaedics, West China Hospital, Sichuan University , Chengdu, China .
authors.forename: Li
authors.initials: L
authors.lastname: Min
authors.affiliation: 2 Department of Orthopedics, The Affiliated Hospital of Zunyi Medical College , Guizhou, China .
authors.forename: Song
authors.initials: S
authors.lastname: Hong
authors.affiliation: 1 Department of Orthopaedics, West China Hospital, Sichuan University , Chengdu, China .
authors.forename: Hong
authors.initials: H
authors.lastname: Duan
authors.affiliation: 1 Department of Orthopaedics, West China Hospital, Sichuan University , Chengdu, China .
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: 1 Department of Orthopaedics, West China Hospital, Sichuan University , Chengdu, China .
authors.forename: Wenli
authors.initials: W
authors.lastname: Zhang
authors.affiliation: 1 Department of Orthopaedics, West China Hospital, Sichuan University , Chengdu, China .
authors.forename: Yi
authors.initials: Y
authors.lastname: Luo
authors.affiliation: 1 Department of Orthopaedics, West China Hospital, Sichuan University , Chengdu, China .
authors.forename: Rui
authors.initials: R
authors.lastname: Shi
authors.affiliation: 1 Department of Orthopaedics, West China Hospital, Sichuan University , Chengdu, China .
authors.forename: Chongqi
authors.initials: C
authors.lastname: Tu
mesh_terms: D017209:Apoptosis / Q000235:genetics; D001859:Bone Neoplasms / Q000235:genetics / Q000473:pathology*; D002453:Cell Cycle / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics*; D049109:Cell Proliferation / Q000235:genetics*; D006801:Humans; D012516:Osteosarcoma / Q000235:genetics / Q000473:pathology*; D062085:RNA, Long Noncoding / Q000037:antagonists & inhibitors / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
chemical_list: C571057:ANCR long noncoding RNA, human; D062085:RNA, Long Noncoding; D012333:RNA, Messenger; D034741:RNA, Small Interfering
keywords: antidifferentiation; lentivirus; noncoding RNA; osteosarcoma; proliferation