title: Overexpression of TIMP-1 mediated by recombinant adenovirus in hepatocellular carcinoma cells inhibits proliferation and invasion in vitro.
abstract: BACKGROUND
  Matrix metalloproteinases (MMPs) and its natural tissue inhibitors of metalloproteinases (TIMPs) are involved in cancer progression. This study was undertaken to determine the effects of overexpression of TIMP-1 on human hepatocellular carcinoma (HCC) cell growth, proliferation, and invasion.
  
  
  METHODS
  Employing the efficient AdEasy(TM) system, recombinant adenovirus AdTIMP-1 containing full-length cDNA of TIMP-1 was generated by homologous recombination and amplified in 293 cells. Then, human HCC cell line (HepG2) underwent gene transfection to overexpress TIMP-1 (so-called HepG-T cells). The mRNA and protein expressions of TIMP-1 were detected with RT-PCR and Western blotting, respectively. The ultrastructure was observed with a transmission electron microscope and the proliferation of HepG-T cells was determined by MTT assay and growth curve. The potential of in vitro invasion was measured with Millicell Chamber.
  
  
  RESULTS
  The resulting AdTIMP-1 and HepG-T cells were generated and the expression of TIMP-1 was detected in vitro. The cell proliferation curves and MTT assay showed HepG-T cells' growth, and proliferation were obviously inhibited. The invasion across Matrigel-coated filters was significantly decreased compared with controls. The suppression rate of HepG-2 cells with AdhTIMP-1 transfection was 50%, and AdhTIMP-1 transfection inhibited by more than 91.6% of the invasion into the Matrigel-coated filter (P<0.01).
  
  
  CONCLUSIONS
  TIMP-1 overexpression results in the suppression of proliferative and invasive potential of HepG2 cells in vitro. This study demonstrates the potential role of TIMP-1 as a target for liver cancer gene therapy and has laid a foundation for further study on its anticancer function.
authors.affiliation: Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Dong
authors.initials: D
authors.lastname: Xia
authors.forename: Lu-Nan
authors.initials: LN
authors.lastname: Yan
authors.forename: Jian-Guo
authors.initials: JG
authors.lastname: Xie
authors.forename: Yu
authors.initials: Y
authors.lastname: Tong
authors.forename: Mao-Lin
authors.initials: ML
authors.lastname: Yan
authors.forename: Xin-Ping
authors.initials: XP
authors.lastname: Wang
authors.forename: Ming-Man
authors.initials: MM
authors.lastname: Zhang
authors.forename: Lan-Ying
authors.initials: LY
authors.lastname: Zhao
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000502:physiology*; D001483:Base Sequence; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D017931:DNA Primers; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism* / Q000473:pathology; D046529:Microscopy, Electron, Transmission; D009361:Neoplasm Invasiveness; D012333:RNA, Messenger / Q000235:genetics; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger; D019715:Tissue Inhibitor of Metalloproteinase-1