Advertisement

Effect of an adenoviral vector that expresses the canine p53 gene on cell growth of canine osteosarcoma and mammary adenocarcinoma cell lines

Mitsuhiro YazawaDepartment of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.

Search for other papers by Mitsuhiro Yazawa in
Current site
Google Scholar
PubMed
Close
 DVM
,
Asuka SetoguchiDepartment of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.

Search for other papers by Asuka Setoguchi in
Current site
Google Scholar
PubMed
Close
 DVM, PhD
,
Sung-Hyeok HongDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.

Search for other papers by Sung-Hyeok Hong in
Current site
Google Scholar
PubMed
Close
 DVM, PhD
,
Rina UyamaDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.

Search for other papers by Rina Uyama in
Current site
Google Scholar
PubMed
Close
 DVM, PhD
,
Takayuki NakagawaDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.

Search for other papers by Takayuki Nakagawa in
Current site
Google Scholar
PubMed
Close
 DVM
,
Noriko KanayaDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.

Search for other papers by Noriko Kanaya in
Current site
Google Scholar
PubMed
Close
 DVM
,
Ryohei NishimuraDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.

Search for other papers by Ryohei Nishimura in
Current site
Google Scholar
PubMed
Close
 DVM, PhD
,
Nobuo SasakiDepartment of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.

Search for other papers by Nobuo Sasaki in
Current site
Google Scholar
PubMed
Close
 DVM, PhD
,
Kenichi MasudaDepartment of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.

Search for other papers by Kenichi Masuda in
Current site
Google Scholar
PubMed
Close
 DVM
,
Koichi OhnoDepartment of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.

Search for other papers by Koichi Ohno in
Current site
Google Scholar
PubMed
Close
 DVM, PhD
, and
Hajime TsujimotoDepartment of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan.

Search for other papers by Hajime Tsujimoto in
Current site
Google Scholar
PubMed
Close
 DVM, PhD

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.

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 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 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-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- 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 functional p53 gene. AxCA-cp53 may be useful to target the p53 gene in the treatment of dogs with tumors. (Am J Vet Res 2003;64:880–888)

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.

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 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 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-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- 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 functional p53 gene. AxCA-cp53 may be useful to target the p53 gene in the treatment of dogs with tumors. (Am J Vet Res 2003;64:880–888)