title: Antitumoral gene-based strategy involving nitric oxide synthase type III overexpression in hepatocellular carcinoma.
abstract: Hepatocellular carcinoma develops in cirrhotic liver. The nitric oxide (NO) synthase type III (NOS-3) overexpression induces cell death in hepatoblastoma cells. The study developed gene therapy designed to specifically overexpress NOS-3 in cultured hepatoma cells, and in tumors derived from orthotopically implanted tumor cells in fibrotic livers. Liver fibrosis was induced by CCl4 administration in mice. The first-generation adenoviruses were designed to overexpress NOS-3 or green fluorescent protein, and luciferase complementary DNA under the regulation of murine alpha-fetoprotein (AFP) and Rous Sarcoma Virus (RSV) promoters, respectively. Both adenovirus and Hepa 1-6 cells were used for in vitro and in vivo experiments. Adenoviruses were administered through the tail vein 2 weeks after orthotopic tumor cell implantation. AFP-NOS-3/RSV-luciferase increased oxidative-related DNA damage, p53, CD95/CD95L expression and caspase-8, -9 and -3 activities in cultured Hepa 1-6 cells. The increased expression of CD95/CD95L and caspase-8 activity was abolished by Nω-nitro-l-arginine methyl ester hydrochloride, p53 and CD95 small interfering RNA. AFP-NOS-3/RSV-luciferase adenovirus increased cell death markers, and reduced cell proliferation of established tumors in fibrotic livers. The increase of oxidative/nitrosative stress induced by NOS-3 overexpression induced DNA damage, p53, CD95/CD95L expression and cell death in hepatocellular carcinoma cells. The effectiveness of the gene therapy has been demonstrated in vitro and in vivo.
authors.affiliation: Oncology Surgery, Cell Therapy and Transplant Organs, Institute of Biomedicine of Seville (IBiS), 'Virgen del Rocío'-'Virgen Macarena' University Hospital/Universidad de Sevilla/CSIC, Sevilla, Spain.
authors.forename: Á J
authors.initials: ÁJ
authors.lastname: De la Rosa
authors.affiliation: Oncology Surgery, Cell Therapy and Transplant Organs, Institute of Biomedicine of Seville (IBiS), 'Virgen del Rocío'-'Virgen Macarena' University Hospital/Universidad de Sevilla/CSIC, Sevilla, Spain.
authors.forename: Á
authors.initials: Á
authors.lastname: Rodríguez-Hernández
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Cordoba, Instituto Maimónides de Investigación Biomédica de Córdoba (IMIBIC), Córdoba, Spain.
authors.forename: R
authors.initials: R
authors.lastname: González
authors.affiliation: Mayo Clinic College of Medicine, Rochester, MN, USA.
authors.forename: S
authors.initials: S
authors.lastname: Romero-Brufau
authors.affiliation: Oncology Surgery, Cell Therapy and Transplant Organs, Institute of Biomedicine of Seville (IBiS), 'Virgen del Rocío'-'Virgen Macarena' University Hospital/Universidad de Sevilla/CSIC, Sevilla, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Navarro-Villarán
authors.affiliation: Department of General Surgery, "Virgen del Rocío"-"Virgen Macarena" University Hospital/IBiS/CSIC/Universidad de Sevilla, Sevilla, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Barrera-Pulido
authors.affiliation: Oncology Surgery, Cell Therapy and Transplant Organs, Institute of Biomedicine of Seville (IBiS), 'Virgen del Rocío'-'Virgen Macarena' University Hospital/Universidad de Sevilla/CSIC, Sevilla, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Pereira
authors.affiliation: Department of General Surgery, "Virgen del Rocío"-"Virgen Macarena" University Hospital/IBiS/CSIC/Universidad de Sevilla, Sevilla, Spain.
authors.forename: L M
authors.initials: LM
authors.lastname: Marín
authors.affiliation: Department of General Surgery, "Virgen del Rocío"-"Virgen Macarena" University Hospital/IBiS/CSIC/Universidad de Sevilla, Sevilla, Spain.
authors.forename: F
authors.initials: F
authors.lastname: López-Bernal
authors.affiliation: Department of General Surgery, "Virgen del Rocío"-"Virgen Macarena" University Hospital/IBiS/CSIC/Universidad de Sevilla, Sevilla, Spain.
authors.forename: J M
authors.initials: JM
authors.lastname: Álamo
authors.affiliation: Department of General Surgery, "Virgen del Rocío"-"Virgen Macarena" University Hospital/IBiS/CSIC/Universidad de Sevilla, Sevilla, Spain.
authors.forename: M A
authors.initials: MA
authors.lastname: Gómez-Bravo
authors.affiliation: Department of General Surgery, "Virgen del Rocío"-"Virgen Macarena" University Hospital/IBiS/CSIC/Universidad de Sevilla, Sevilla, Spain.
authors.forename: F J
authors.initials: FJ
authors.lastname: Padillo
authors.affiliation: Department of General Surgery, "Virgen del Rocío"-"Virgen Macarena" University Hospital/IBiS/CSIC/Universidad de Sevilla, Sevilla, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Muntané
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000628:therapy*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D053181:Caspase 8 / Q000235:genetics / Q000378:metabolism; D053453:Caspase 9 / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004249:DNA Damage; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D053222:Fas Ligand Protein / Q000235:genetics / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008103:Liver Cirrhosis / Q000235:genetics / Q000628:therapy; D008113:Liver Neoplasms / Q000235:genetics / Q000628:therapy*; D051379:Mice; D019331:NG-Nitroarginine Methyl Ester / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D052736:Rous sarcoma virus / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism; D000509:alpha-Fetoproteins / Q000235:genetics / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D053222:Fas Ligand Protein; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; D000509:alpha-Fetoproteins; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse; C505409:Casp3 protein, mouse; C505526:Casp8 protein, mouse; C506434:Casp9 protein, mouse; D053148:Caspase 3; D053181:Caspase 8; D053453:Caspase 9; D019331:NG-Nitroarginine Methyl Ester