title: A hammerhead ribozyme suppresses expression of hepatocyte growth factor/scatter factor receptor c-MET and reduces migration and invasiveness of breast cancer cells.
abstract: OBJECTIVE
  Hepatocyte growth factor/scatter factor (HGF/SF), via its receptor c-MET, has been implicated to play a pivotal role in breast cancer development and progression. This study examined a transgene-consisting of a combination of U1snRNA, hammerhead ribozyme, and antisense, designed to inhibit c-met expression-and its impact on the migration and in vitro invasion of breast cancer cells.
  
  
  METHODS
  A hammerhead ribozyme targeting human c-MET was cloned into a modified pZeoU1EcoSpe vector and transfected into breast cancer cells MDA MB 231 and MCF-7 by electroporation. Expression of MET mRNA and protein was determined. Migration and in vitro invasiveness of transfected cells were also analyzed.
  
  
  RESULTS
  Breast cancer cells were transfected with the ribozyme-containing plasmids. Stable transfectants manifested an almost complete loss of MET mRNA and protein, as shown by reverse transcription-PCR, Northern blotting, and Western blotting, respectively, whereas the wild-type plasmid had no effects. Met-ribozyme transfected cells exhibited reduced migration and in vitro invasiveness through extracellular matrix (Matrigel), compared with the wild-type cells and cells transfected with empty plasmid.
  
  
  CONCLUSIONS
  These data show that targeting c-MET by way of a hammerhead ribozyme encoding antisense to c-MET is an effective approach in reducing the invasiveness of breast cancer cells.
authors.affiliation: University Department of Surgery, University of Wales College of Medicine, Cardiff CF14 4XN, United Kingdom. jiangw@cf.ac.uk
authors.forename: W G
authors.initials: WG
authors.lastname: Jiang
authors.forename: D
authors.initials: D
authors.lastname: Grimshaw
authors.forename: J
authors.initials: J
authors.lastname: Lane
authors.forename: T A
authors.initials: TA
authors.lastname: Martin
authors.forename: R
authors.initials: R
authors.lastname: Abounader
authors.forename: J
authors.initials: J
authors.lastname: Laterra
authors.forename: R E
authors.initials: RE
authors.lastname: Mansel
authors.forename: R
authors.initials: R
authors.lastname: Abounder
mesh_terms: D001483:Base Sequence; D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology* / Q000628:therapy; D002465:Cell Movement / Q000235:genetics; D016373:DNA, Antisense / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008969:Molecular Sequence Data; D009361:Neoplasm Invasiveness / Q000235:genetics / Q000517:prevention & control; D010957:Plasmids / Q000235:genetics; D019859:Proto-Oncogene Proteins c-met / Q000235:genetics* / Q000378:metabolism; D016337:RNA, Catalytic / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012342:RNA, Small Nuclear / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D016373:DNA, Antisense; D016337:RNA, Catalytic; D012333:RNA, Messenger; D012342:RNA, Small Nuclear; C405483:U1 small nuclear RNA; D019859:Proto-Oncogene Proteins c-met