Nitric Oxide Synthase Type III Overexpression By Gene Therapy Exerts Antitumoral Activity In Mouse Hepatocellular Carcinoma.
Hepatocellular carcinoma develops in cirrhotic liver. The nitric oxide (NO) synthase type III (NOS-3) overexpression induces cell death in hepatoma 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. Hepa 1-6 cells were used for in vitro and in vivo experiments. The first generation adenovirus was designed to overexpress NOS-3 (or GFP) and luciferase cDNA under the regulation of murine alpha-fetoprotein (AFP) and Rous Sarcoma Virus (RSV) promoters, respectively. Both adenoviruses were administered through the tail vein two weeks after orthotopic tumor cell implantation. AFP-NOS-3/RSV-Luciferase increased oxidative-related DNA damage, p53, CD95/CD95L expression and caspase-8 activity in cultured Hepa 1-6 cells. The increased expression of CD95/CD95L and caspase-8 activity was abolished by l-NAME or p53 siRNA. The tail vein infusion of 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.
Departament of Biochemistry and Molecular Biology, Universidad de Córdoba, Instituto Maimónides de Investigación Biomédica de Córdoba (IMIBIC), Spain.
Raúl
R
González
Cirugía Oncológica, Terapia Celular y Trasplante de Órganos, Instituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío-Virgen Macarena/Universidad de Sevilla/CSIC, Spain.
Ángel J
ÁJ
De la Rosa
Mayo Clinic College of Medicine, Rochester, USA.
Santiago
S
Romero-Brufau
Department of General Surgery, Hospital Universitario Virgen del Rocío-Virgen Macarena/IBiS/CSIC/Universidad de Sevilla, Spain.
Lydia
L
Barrera-Pulido
Cirugía Oncológica, Terapia Celular y Trasplante de Órganos, Instituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío-Virgen Macarena/Universidad de Sevilla/CSIC, Spain.
Francisco
F
Gallardo-Chamizo
Cirugía Oncológica, Terapia Celular y Trasplante de Órganos, Instituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío-Virgen Macarena/Universidad de Sevilla/CSIC, Spain.
Sheila
S
Pereira
Department of General Surgery, Hospital Universitario Virgen del Rocío-Virgen Macarena/IBiS/CSIC/Universidad de Sevilla, Spain.
Luís M
LM
Marín
Department of General Surgery, Hospital Universitario Virgen del Rocío-Virgen Macarena/IBiS/CSIC/Universidad de Sevilla, Spain.
José M
JM
Álamo
Cirugía Oncológica, Terapia Celular y Trasplante de Órganos, Instituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío-Virgen Macarena/Universidad de Sevilla/CSIC, Spain.
Ángeles
Á
Rodríguez-Hernández
Department of General Surgery, Hospital Universitario Virgen del Rocío-Virgen Macarena/IBiS/CSIC/Universidad de Sevilla, Spain.
Francisco J
FJ
Padillo
Department of General Surgery, Hospital Universitario Virgen del Rocío-Virgen Macarena/IBiS/CSIC/Universidad de Sevilla, Spain.
Jordi
J
Muntané
D000256:Adenoviridae; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000201:enzymology / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D015316:Genetic Therapy; D008114:Liver Neoplasms, Experimental / Q000201:enzymology / Q000235:genetics / Q000628:therapy*; D051379:Mice; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000096:biosynthesis / Q000235:genetics
D009363:Neoplasm Proteins; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse