title: [Effects of CTCF on human liver cancer stem cells and cell proliferation].
abstract: OBJECTIVE
  To explore the effects of CCCTC-binding factor (CTCF) on human liver cancer stem cells (HepG2) and cell proliferation of HepG2 and Nasopharyngeal carcinoma cell line (CNE1).
  
  
  METHODS
  The pEGFP-N1/CTCF, CTCF-shRNA and GFP-shRNA plasmids were constructed and transfected into HepG2 and CNE1 cells, and RT-PCR or Western blot were performed to detect the mRNA or protein levels of CTCF. The subpopulation of CD90+ cancer stem cells in HepG2 cells transfected with CTCF-shRNA plasmid or GFP-shRNA plasmid (as transfection control) were assayed by flow cytometry with the wild type HepG2 cells as control. Proliferation of cells transfected with CTCF-overexpression or CTCF-shRNA plasmid was evaluated by MTT assay.
  
  
  RESULTS
  The levels of both mRNA and protein of CTCF were increased in pEGFP-N1/CTCF transfected HepG2 and CNE1 cells compared to that in pEGFP-N1 transfected cells (P < 0.05), and decreased in CTCF-shRNA transfected cells compared to that in cells transfected with GFP-shRNA (P < 0.05). The results of flow cytometry demonstrated that, detection rate of CD90+ cells in cells transfected with CTCF-shRNA plasmid [(1.7330 +/- 0.4177)%] was obviously higher than that of wild-type HepG2 cells [(0.5750 +/- 0.0629)%] and cells transfected with GFP-shRNA plasmid [(0.3500 +/- 0.0866)%] (P < 0.05). The results of MTT analysis showed that, alteration of CTCF had no effect on cancer cell proliferation (P > 0.05).
  
  
  CONCLUSIONS
  CTCF inhibits human liver cancer stem cells but no effect on cell proliferation.
authors.forename: Qiu-Ying
authors.initials: QY
authors.lastname: Liu
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Xie
authors.forename: Ling
authors.initials: L
authors.lastname: Wei
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.forename: Wen-Yan
authors.initials: WY
authors.lastname: Shen
authors.forename: Ran
authors.initials: R
authors.lastname: Li
authors.forename: Xiao Qin
authors.initials: XQ
authors.lastname: Yu
authors.forename: Yang
authors.initials: Y
authors.lastname: Qin
mesh_terms: D000076246:CCCTC-Binding Factor; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005434:Flow Cytometry; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism*; D014411:Neoplastic Stem Cells / Q000166:cytology*; D010957:Plasmids; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012097:Repressor Proteins / Q000378:metabolism*; D014162:Transfection
chemical_list: D000076246:CCCTC-Binding Factor; C000619697:CTCF protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012097:Repressor Proteins