Evaluation of an in vitro telomeric repeat amplification protocol assay to detect telomerase activity in canine urine.
OBJECTIVE
To evaluate the usefulness of a PCR-based telomeric repeat amplification protocol (TRAP) assay for detecting telomerase activity in cells from a canine transitional cell carcinoma (TCC) cell line and, ultimately, in the urine of dogs with TCC.


METHODS
11 dogs with histologic or cytologic evidence of TCC, 10 dogs with benign lower urinary tract disease, and 9 healthy dogs.


METHODS
Telomerase activity was initially evaluated in cells from canine TCC (K9TCC) and telomerase-negative (WI-38) cell lines. Following assay optimization, telomerase stability was evaluated at various storage durations and temperatures. Urine samples were then obtained prospectively from study dogs.


RESULTS
Telomerase activity was detected in the K9TCC cell line. The TRAP assay detected telomerase activity in as few as 10 K9TCC cells alone and as low as 2% of a total cell population in K9TCC and WI-38 mixing experiments. A loss of telomerase activity was detected with increasing urine storage durations at various temperatures. Telomerase activity was clearly detected in samples collected from 10 of 11 dogs with TCC, 2 of 10 dogs with benign lower urinary tract disease, and none of the 9 healthy dogs.


CONCLUSIONS
The TRAP-based assay detected telomerase activity in the canine TCC cell line and revealed that the telomerase ribonucleoprotein complex was inherently unstable at various storage durations and conditions. Telomerase activity was also detectable in urine samples obtained from dogs with TCC, which suggested the TRAP assay may be useful in diagnosing TCC in dogs.
Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606, USA.
Angela L
AL
McCleary-Wheeler
Laurel E
LE
Williams
Paul R
PR
Hess
Steven E
SE
Suter
D000818:Animals; D002295:Carcinoma, Transitional Cell / Q000201:enzymology / Q000652:urine / Q000662:veterinary*; D045744:Cell Line, Tumor; D004283:Dog Diseases / Q000201:enzymology* / Q000652:urine; D004285:Dogs / Q000652:urine*; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005347:Fibroblasts / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D016133:Polymerase Chain Reaction / Q000379:methods; D019098:Telomerase / Q000032:analysis / Q000652:urine*; D016615:Telomere / Q000235:genetics*
D019098:Telomerase