title: Inhibitory effect of adeno-associated virus-mediated gene transfer of human endostatin on hepatocellular carcinoma.
abstract: OBJECTIVE
  To investigate the effect of adeno-associated virus-mediated gene transfer of human endostatin on the growth of hepatocellular carcinoma (HCC).
  
  
  METHODS
  HCC cell line Hep3B was infected with recombinant adeno-associated virus containing human endostatin gene (rAAV2-hEndo). The results of transfection were detected by RT-PCR and SDS-PAGE assay. MTT assay was used to observe the effects of supernatant of transfected cells on ECV304 cell proliferation. An animal model of HCC was established by injecting Hep3B cells subcutaneously into the back of nude mice. Intratumoral injection of rAAV2-hEndo, empty virus and phosphate-buffered saline were given sequentially. Serum endostatin was determined by ELISA, the inhibitory effect of endostatin on the growth of xenograft was assessed in 3 wk.
  
  
  RESULTS
  The results of RT-PCR and SDS-PAGE assay confirmed that rAAV2-hEndo successfully transfected Hep3B cells, and endostatin was secreted from Hep3B cells to medium. The supernatant of transfected cells markedly inhibited the proliferation of ECV304 cells (P<0.01). Intratumoral injection of rAAV2-hEndo (2 x 10(10) v.g.) led to a sustained serum endostatin level of approximately (86.71+/-5.19) ng/mL. The tumor volume and microvessel density were less in rAAV2-hEndo group than in control groups (P<0.01).
  
  
  CONCLUSIONS
  Human endostatin can be stably expressed by adeno-associated virus-mediated gene transfer and effectively inhibit the growth of HCC.
authors.affiliation: Department of Surgery, Second Affiliated Hospital, Zhejiang University College of Medicine, 88 Jiefang Road, Hangzhou 310006, Zhejiang Province, China. lhong.lli@sohu.com
authors.forename: Hong
authors.initials: H
authors.lastname: Liu
authors.forename: Cheng-Hong
authors.initials: CH
authors.lastname: Peng
authors.forename: Ying-Bin
authors.initials: YB
authors.lastname: Liu
authors.forename: Yu-Lian
authors.initials: YL
authors.lastname: Wu
authors.forename: Zhi-Ming
authors.initials: ZM
authors.lastname: Zhao
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.forename: Bao-San
authors.initials: BS
authors.lastname: Han
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D043169:Endostatins / Q000235:genetics*; D018014:Gene Transfer Techniques; D006801:Humans; D008113:Liver Neoplasms
chemical_list: D043169:Endostatins