title: [Effect of lentivirus-mediated nestin gene silencing on metastatic potential of human melanoma cell line UACC903 in vitro].
abstract: OBJECTIVE
  To study the effect of lentivirus-mediated RNA interference (RNAi) of nestin on the metastatic potential of human melanoma cell line UACC903.
  
  
  METHODS
  A lentiviral vector for RNAi targeting the coding region of human nestin mRNA (nestin-RNAi-LV) and another control vector containing a nonsense sequence were constructed. The vectors were transfected into UACC903 cells, and nestin expression in the cells was detected by RT-PCR, immunofluorence assay and Western blotting. The invasive ability and migration of the transfected UACC903 cells was evaluated using Transwell and scrape assay, respectively. Fluorescence assay was used to examine the expressions of E-cadherin, N-cadherin and β-catenin in the cells.
  
  
  RESULTS
  The lentiviral vector nestin-RNAi-LV was constructed successfully. Compared with the control vector, nestin-RNAi-LV resulted in obviously reduced expression of nestin mRNA and protein, lowered migration ability of UACC903 cells, and reduced cell adhesion  and invasiveness (P<0.05).
  
  
  CONCLUSIONS
  Lentivirus-mediated nestin RNAi can specifically inhibit nestin expression to cause decreased cell migration and invasiveness of human melanoma cell line UACC903.
authors.affiliation: Guangdong Women and Children's Hospital, Guangzhou, China. yxhshy52@163.com
authors.forename: Xuhui
authors.initials: X
authors.lastname: Yang
authors.forename: Tian
authors.initials: T
authors.lastname: Xia
authors.forename: Weihua
authors.initials: W
authors.lastname: Yu
authors.forename: Xiaofang
authors.initials: X
authors.lastname: Lu
authors.forename: Peng
authors.initials: P
authors.lastname: Xiang
authors.forename: Feng
authors.initials: F
authors.lastname: He
mesh_terms: D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D007381:Intermediate Filament Proteins / Q000235:genetics* / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics*; D008545:Melanoma / Q000473:pathology*; D009362:Neoplasm Metastasis / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D064231:Nestin; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D012333:RNA, Messenger; D034741:RNA, Small Interfering