title: Effect of an adenoviral vector that expresses the canine p53 gene on cell growth of canine osteosarcoma and mammary adenocarcinoma cell lines.
abstract: OBJECTIVE
  To generate an adenoviral vector that expressed the canine p53 gene and investigate its growth-inhibiting effect on canine osteosarcoma and mammary adenocarcinoma cell lines.
  
  
  METHODS
  2 canine osteosarcoma cell lines (HOS, OOS) and 3 canine mammary adenocarcinoma cell lines (CHMp, CIPm, and CNMm).
  
  
  METHODS
  An adenoviral vector that expressed the canine p53 gene (AxCA-cp53) was generated. p53 gene 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 beta-galactosidase (AxCA-LacZ) and 3-indolyl-beta-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-derived p53 gene 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-beta-D-galactopyranoside staining results ranged from 10 to 50. AxCA-cp53 induced growth inhibition in a dose-dependent manner. Arrest of the G1-phase population and apoptotic DNA fragmentation were observed in cells infected with AxCA-cp53.
  
  
  CONCLUSIONS
  AxCA-cp53 inhibits cell growth via induction of cell cycle arrest and apoptosis in canine osteosarcoma and mammary adenocarcinoma cell lines that lack a functional p53 gene. AxCA-cp53 may be useful to target the p53 gene in the treatment of dogs with tumors.
authors.affiliation: Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Yazawa
authors.forename: Asuka
authors.initials: A
authors.lastname: Setoguchi
authors.forename: Sung-Hyeok
authors.initials: SH
authors.lastname: Hong
authors.forename: Rina
authors.initials: R
authors.lastname: Uyama
authors.forename: Takayuki
authors.initials: T
authors.lastname: Nakagawa
authors.forename: Noriko
authors.initials: N
authors.lastname: Kanaya
authors.forename: Ryohei
authors.initials: R
authors.lastname: Nishimura
authors.forename: Nobuo
authors.initials: N
authors.lastname: Sasaki
authors.forename: Kenichi
authors.initials: K
authors.lastname: Masuda
authors.forename: Koichi
authors.initials: K
authors.lastname: Ohno
authors.forename: Hajime
authors.initials: H
authors.lastname: Tsujimoto
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology* / Q000662:veterinary; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017209:Apoptosis; D002453:Cell Cycle; D002455:Cell Division; D004285:Dogs; D005260:Female; D015870:Gene Expression; D016158:Genes, p53 / Q000235:genetics*; D015674:Mammary Neoplasms, Animal / Q000235:genetics / Q000473:pathology*; D012516:Osteosarcoma / Q000235:genetics / Q000473:pathology* / Q000662:veterinary; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: D016159:Tumor Suppressor Protein p53