title: The influence of human single chain inteleukin-12 gene transduction on the biological behavior of hepatoma 7721 cells.
abstract: OBJECTIVE
  To investigate the anti-tumor effects of human single chain interleukin-12 (hscIL-12).
  
  
  METHODS
  pcDNA/hscIL-12 recombinant was transfected into human hepatic carcinoma cells (7721 cells) by lipofectin method. The 7721/hscIL-12 cells which secrete hscIL-12 stably, were obtained via G418 selection, and in vitro the influence of hscIL-12 gene transduction on the growth of tumor cells was evaluated by cell cycle analysis. In vivo, genetically engineered 7721 cells (7721/hscIL-12, 7721/pcDNA) and parental cells were implanted into BALB/c nude mice, respectively. 7721/pcDNA and 7721/hscIL-12 groups were divided into two sub-groups on day 8: one was administered with hPBL twice, 6 days at interval; the other was given equal volume of PBS. Mice were sacrificed on day 26, and spleens and tumors were taken out for histologic assay.
  
  
  RESULTS
  hscIL-12 produced stably by 7721/hscIL-12 cells had bioactivity, and it was proved by Western blot, immunocytochemistry, and in situ hybridization. In vitro, compared with 7721 and 7721/pcDNA, the 7721/hscIL-12 grew much more slowly. FACS assay showed apparent G1 arrest of 7721/hscIL-12 cells. In animal experiment, on day 8 after inoculation, the tumors of 7721 and 7721/pcDNA group were up to 5 approximately 7 mm, while those of 7721/hscIL-12 group were 2 approximately 4 mm. When treated with hPBL, the tumor of 7721/hscIL-12 group disappeared completely. Histologically, the tumors from 7721/hscIL-12 without hPBL treatment had numerous lymphocyte infiltration, the tumor cells displayed depression looking, atrophy, focal necrosis and apoptosis, whereas the tumors of 7721 and 7721/pcDNA groups grew thrivingly.
  
  
  CONCLUSIONS
  hscIL-12 transduced 7721 cells could induced significant antitumor immune response which resulted in tumor regression totally when the hPBL was inoculated, and also hscIL-12 has certain effects on mice immune system. These findings suggest that hscIL-12 and hscIL-12 gene therapy might have promising prospects in clinical application.
authors.affiliation: Department of Immunopathology, Xi'an Medical University, Xi'an 710061. jinli73@yahoo.com
authors.forename: L
authors.initials: L
authors.lastname: Jin
authors.forename: B
authors.initials: B
authors.lastname: Lai
authors.forename: Y
authors.initials: Y
authors.lastname: Geng
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.forename: L
authors.initials: L
authors.lastname: Si
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D002455:Cell Division; D015316:Genetic Therapy; D006801:Humans; D018664:Interleukin-12 / Q000096:biosynthesis / Q000235:genetics*; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D018664:Interleukin-12