Effect of an adenoviral vector that expresses the canine p53 gene on cell growth of canine osteosarcoma and mammary adenocarcinoma cell lines
| Autor: | Nobuo Sasaki, Ryohei Nishimura, Noriko Kanaya, Mitsuhiro Yazawa, Kenichi Masuda, Rina Uyama, Takayuki Nakagawa, Hajime Tsujimoto, Koichi Ohno, Asuka Setoguchi, Sung-Hyeok Hong |
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| Rok vydání: | 2003 |
| Předmět: |
Cell
Gene Expression Apoptosis Mammary Neoplasms Animal Adenocarcinoma Biology Canine Osteosarcoma Adenoviridae Viral vector Flow cytometry Dogs Gene expression Tumor Cells Cultured medicine Animals Osteosarcoma General Veterinary medicine.diagnostic_test Cell growth Cell Cycle General Medicine Cell cycle Genes p53 Molecular biology medicine.anatomical_structure Cell culture Female Tumor Suppressor Protein p53 Cell Division |
| Zdroj: | American Journal of Veterinary Research. 64:880-888 |
| ISSN: | 0002-9645 |
| Popis: | Objective—To generate an adenoviral vector that expressed the caninep53gene and investigate its growth-inhibiting effect on canine osteosarcoma and mammary adenocarcinoma cell lines.Sample Population—2 canine osteosarcoma cell lines (HOS, OOS) and 3 canine mammary adenocarcinoma cell lines (CHMp, CIPm, and CNMm).Procedure—An adenoviral vector that expressed the caninep53gene (AxCA-cp53) was generated.p53gene expression was examined by use of reverse transcription (RT)-polymerase chain reaction (PCR) assay and immunohistochemistry. Susceptibility of cell lines to the adenoviral vector was determined by infection with an adenoviral vector that expresses β-galactosidase (AxCA-LacZ) and 3-indolyl-β-D-galactopyranoside staining. Growth inhibitory effects were examined by monitoring the numbers of cells after infection with mock (PBS) solution, AxCA-LacZ, or AxCA-cp53. The DNA contents per cell were measured by flow cytometry analysis. Apoptotic DNA fragmentation was detected by use of a terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay.Results—AxCA-cp53-derivedp53gene mRNA and P53 protein were detected by RT-PCR analysis and immunohistochemistry, respectively. Multiplicity of infection at which 50% of cells had positive 3-indolyl- β-D-galactopyranoside staining results ranged from 10 to 50. AxCA-cp53 induced growth inhibition in a dosedependent manner. Arrest of the G1-phase population and apoptotic DNA fragmentation were observed in cells infected with AxCA-cp53.Conclusions and Clinical Relevance—AxCA-cp53 inhibits cell growth via induction of cell cycle arrest and apoptosis in canine osteosarcoma and mammary adenocarcinoma cell lines that lack a functionalp53gene. AxCA-cp53 may be useful to target thep53gene in the treatment of dogs with tumors. (Am J Vet Res2003;64:880–888) |
| Databáze: | OpenAIRE |
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