title: [Lentivirus-mediated RNA interference of CD133 inhibits the proliferation of CD133(+) liver cancer stem cells and increases their cisplatin chemosensitivity].
abstract: OBJECTIVE
  To study the effects of CD133 suppression by lentivirus-mediated RNA interference (RNAi) on the proliferation and chemosensitivity of CD133(+) cancer stem cells (CSCs) sorted from HepG2 cell line.
  
  
  METHODS
  CD133(+) and CD133- cells were sorted from HepG2 cell line by flow cytometry, and the expression of CD133 before and after cell sorting were detected. The stem cell property of sorted CD133(+) cells were validated by sphere-forming assay in vitro and xenograft experiments in vivo. Lentivirus-mediated short hairpin RNA (shRNA) targeting CD133 were transfected into CD133(+) cells, and CD133 mRNA and protein expressions of the transfected cells were detected by RT-PCR and Western blotting, respectively. Before and after the transfection, the proliferative ability of CD133(+) cells was evaluated by colony formation assay, and the cell growth inhibition rate and apoptosis following cisplatin exposure were detected using CCK-8 assay and flow cytometry.
  
  
  RESULTS
  The sorted CD133(+) cells showed a high purity of (88.74∓3.19)%, as compared with the purity of (3.36∓1.80)% before cell sorting. CD133(+) cells showed a high tumor sphere formation ability and tumorigenesis capacity compared with CD133- cells. CD133 shRNA transfection significantly inhibited CD133 mRNA and protein expressions in CD133(+) cells (P<0.01), resulting also in a significantly lowered cell proliferative ability (P<0.01) and an increased growth inhibition rate (P<0.01) and obviously increased cell apoptosis (P<0.05) after cisplatin exposure.
  
  
  CONCLUSIONS
  Lentivirus-mediated RNAi for CD133 suppression inhibits the proliferation of CD133(+) liver cancer stem cells and increases their chemosensitivity to cisplatin.
authors.affiliation: Department of Radiology, Chongqing Medical University, Chongqing 400016, China. viviankuku@163.com
authors.forename: Xi
authors.initials: X
authors.lastname: Lan
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.forename: Shu
authors.initials: S
authors.lastname: Cao
authors.forename: Dongling
authors.initials: D
authors.lastname: Zou
authors.forename: Fang
authors.initials: F
authors.lastname: Li
authors.forename: Shaolin
authors.initials: S
authors.lastname: Li
mesh_terms: D000071916:AC133 Antigen; D015703:Antigens, CD / Q000235:genetics*; D049109:Cell Proliferation; D002945:Cisplatin / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm; D006023:Glycoproteins / Q000235:genetics*; D056945:Hep G2 Cells; D006801:Humans; D016086:Lentivirus; D014411:Neoplastic Stem Cells / Q000166:cytology* / Q000187:drug effects*; D010455:Peptides / Q000235:genetics*; D034622:RNA Interference
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D002945:Cisplatin