title: [Pigment epithelium-derived factor gene therapy inhibits the growth of transplanted human hepatocellular carcinoma in nude mice].
abstract: OBJECTIVE
  To evaluate the antiangiogenic property of pigment epithelium-derived factor(PEDF) in heptocarcinoma cell lines and explore its possible application in the gene therapy of human hepatocellular carcinoma (HCC).
  
  
  METHODS
  The gene encoding human PEDF was subcloned into lentiviral vector to generate the recombinant plasmid pLenti-PEDF. The plasmid pLenti-PEDF and two other packaging plasmids were cotransfected to 293T cells by calcium phosphate. Then HepG2 was infected with recombinant lentivirus and the expression efficiency of PEDF was analyzed by western blot. Proliferation and migration assay of human umbilical vein endothelial cells (HUVEC) was used to evaluate the biological activity of PEDF in vitro. Murine subcutaneous tumor model was established to investigate the therapeutic effects of Lenti-PEDF on HCC, and the expression of PEDF mRNA in tumor tissues was analyzed by RT-PCR.
  
  
  RESULTS
  Restriction enzyme digestion and DNA sequencing demonstrated that the recombinant plasmid pLenti-PEDF was constructed successfully. HepG2 secreted PEDF in the media effectively after infected with the recombinant lentivirus and this protein exhibited strong inhibitory effects on proliferation and migration of human umbilical vein endothelial cells (P less than 0.01). Intratumoral injection of Lenti-PEDF caused significant inhibition of tumor growth (P less than 0.01), and high level expression of PEDF mRNA was detected in tumor tissues by RT-PCR.
  
  
  CONCLUSIONS
  Our data suggest that PEDF may exert an inhibitory effect on tumor angiogenesis and PEDF gene therapy may provide a new approach for the treatment of HCC.
authors.affiliation: Institute of Liver Transplantation, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: GAO
authors.forename: Wei
authors.initials: W
authors.lastname: ZHANG
authors.forename: Liang
authors.initials: L
authors.lastname: ZHAO
authors.forename: Xue-hao
authors.initials: XH
authors.lastname: WANG
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D042783:Endothelial Cells / Q000378:metabolism; D005136:Eye Proteins / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy; D005822:Genetic Vectors; D056945:Hep G2 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008113:Liver Neoplasms / Q000098:blood supply / Q000235:genetics / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009389:Neovascularization, Pathologic / Q000628:therapy*; D009414:Nerve Growth Factors / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015843:Serpins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology; D000099174:Pigment Epithelium-Derived Factor
chemical_list: D005136:Eye Proteins; D009414:Nerve Growth Factors; D012333:RNA, Messenger; D015843:Serpins; D000099174:Pigment Epithelium-Derived Factor