Antitumoral gene-based strategy involving nitric oxide synthase type III overexpression in hepatocellular carcinoma

• Published in: Gene therapy 2016-01, Vol.23 (1), p.67-77
• Main Authors: De la Rosa, Á J, Rodríguez-Hernández, Á, González, R, Romero-Brufau, S, Navarro-Villarán, E, Barrera-Pulido, L, Pereira, S, Marín, L M, López-Bernal, F, Álamo, J M, Gómez-Bravo, M A, Padillo, F J, Muntané, J
• Format: Article
• Language: English
• Subjects: 631/61/2300/1514; 631/61/51/201; 692/4028/67/1504/1610/4029; 692/699/67/1504/1610; Adenoviridae - genetics; Adenovirus; Adenoviruses; alpha-Fetoproteins - genetics; alpha-Fetoproteins - metabolism; Animals; Apoptosis; Arginine; Biomedicine; Carbon tetrachloride; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - therapy; Care and treatment; Caspase 3 - genetics; Caspase 3 - metabolism; Caspase 8 - genetics; Caspase 8 - metabolism; Caspase 9 - genetics; Caspase 9 - metabolism; Caspase-8; CD95 antigen; Cell Biology; Cell death; Cell Line, Tumor; Cell Proliferation; Complementary DNA; Development and progression; Disease Models, Animal; DNA Damage; DNA, Complementary - genetics; DNA, Complementary - metabolism; Fas Ligand Protein - genetics; Fas Ligand Protein - metabolism; FasL protein; Fibrosis; Gene Expression; Gene Expression Regulation, Neoplastic; Gene Therapy; Genetic aspects; Genetic Therapy - methods; Genetic Vectors; Green fluorescent protein; Health aspects; Hepatocellular carcinoma; Hepatoma; Human Genetics; Liver; Liver - cytology; Liver - metabolism; Liver cancer; Liver cirrhosis; Liver Cirrhosis - genetics; Liver Cirrhosis - therapy; Liver Neoplasms - genetics; Liver Neoplasms - therapy; Methods; Mice; Nanotechnology; NG-Nitroarginine Methyl Ester - metabolism; Nitric oxide; Nitric Oxide Synthase Type III - genetics; Nitric Oxide Synthase Type III - metabolism; Nitric-oxide synthase; original-article; Oxidoreductases; p53 Protein; Patient outcomes; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; Rous sarcoma virus; Rous sarcoma virus - genetics; siRNA; Tumor cells; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; Tumors; α-Fetoprotein
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/gt.2015.79

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 CCl 4 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 .