Results of hyperamplification of centrosomes in naturally developing tumors of dogs.
OBJECTIVE
To evaluate results of centrosome hyperamplification in naturally developing tumors of dogs.


METHODS
Tumor specimens from 9 dogs with tumors (rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myxosarcoma, and mammary gland tumor) and 2 canine osteosarcoma cell lines.


METHODS
3 antibodies for centrosome proteins (ie, anti-gamma-tubulin, anti-BRCA1, and anti-pericentrin) were used for immunohistochemical analysis. Double immunostaining for centrosomes was used to confirm the specificity of these antibodies for centrosomes. Mutational analysis of the canine p53 gene was carried out by polymerase chain reaction-single-strand conformation polymorphism analysis, and expression of canine MDM2 protein was evaluated by use of immunohistochemical analysis, using anti-MDM2 antibody.


RESULTS
Immunohistochemical analysis of dog osteosarcoma cell lines with apparent aneuploidy revealed frequent hyperamplification of centrosomes in the osteosarcoma cell lines. Similar hyperamplified centrosomes were detected in the tumor tissues from all of the 9 tumors. The frequency of cells with hyperamplified centrosomes (3 to 20/cell) in each tumor tissue ranged from 9.50 to 48.1%, whereas centrosome hyperamplification was not observed in normal lymph nodes from these dogs. In 8 of the 9 tumors, mutation of p53 gene or overexpression of MDM2, or both, was detected.


CONCLUSIONS
Various types of naturally developing tumors in dogs often have hyperamplification of centrosomes associated with chromosome instability. Hyperamplification of centrosomes is a novel tumor marker for use in cytologic and histologic examinations of clinical specimens obtained from dogs.
Setoguchi
A
A
Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, University of Tokyo, Japan.
Okuda
M
M
Nishida
E
E
Yazawa
M
M
Ishizaka
T
T
Hong
S H
SH
Hisasue
M
M
Nishimura
R
R
Sasaki
N
N
Yoshikawa
Y
Y
Masuda
K
K
Ohno
K
K
Tsujimoto
H
H
D000818:Animals; D018385:Centrosome / Q000473:pathology*; D002869:Chromosome Aberrations / Q000662:veterinary*; D004273:DNA, Neoplasm / Q000737:chemistry; D004283:Dog Diseases / Q000235:genetics* / Q000473:pathology; D004285:Dogs; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D016158:Genes, p53 / Q000235:genetics; D007150:Immunohistochemistry / Q000662:veterinary; D008297:Male; D015674:Mammary Neoplasms, Animal / Q000235:genetics* / Q000473:pathology; D009687:Nuclear Proteins; D018807:Polymorphism, Single-Stranded Conformational; D011518:Proto-Oncogene Proteins / Q000032:analysis / Q000096:biosynthesis; D051736:Proto-Oncogene Proteins c-mdm2; D012334:RNA, Neoplasm / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012509:Sarcoma / Q000737:chemistry / Q000235:genetics / Q000662:veterinary*; D017422:Sequence Analysis, DNA; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D012334:RNA, Neoplasm; D051736:Proto-Oncogene Proteins c-mdm2