Nitric Oxide Synthase Type III Overexpression By Gene Therapy Exerts Antitumoral Activity In Mouse Hepatocellular Carcinoma

• Published in: Redox biology Vol. 5; pp. 420 - 421
• Main Authors: González, Raúl, De la Rosa, Ángel J, Romero-Brufau, Santiago, Barrera-Pulido, Lydia, Gallardo-Chamizo, Francisco, Pereira, Sheila, Marín, Luís M, Álamo, José M, Rodríguez-Hernández, Ángeles, Padillo, Francisco J, Muntané, Jordi
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier 01-08-2015
• Subjects: Adenoviridae; Animals; Carcinoma, Hepatocellular - enzymology; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - pathology; Carcinoma, Hepatocellular - therapy; Genetic Therapy; Liver Neoplasms, Experimental - enzymology; Liver Neoplasms, Experimental - genetics; Liver Neoplasms, Experimental - therapy; Mice; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; Nitric Oxide Synthase Type III - biosynthesis; Nitric Oxide Synthase Type III - genetics
• ISSN: 2213-2317; 2213-2317
• DOI: 10.1016/j.redox.2015.09.032

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.