[Telomerase activity and determination of cancer in urological neoplasias using exfoliated urinary cells: in situ TRAP assay and its application].
We determined telomerase activity semiquantitatively and in-cell telomerase activity in exfoliated urinary cells obtained from urological neoplasias. Semiquantitative determination of telomerase activity was performed using a fluorescent-based telomeric repeat amplification protocol (TRAP) and telomerase activity at the cellular level was determined by an in situ TRAP assay. Fluorescent-based TRAP assay detected urinary telomerase activity in about 80% patients with urological neoplasia before treatment, whereas urinary cells obtained from 30% patients after/during treatment with either chemotherapy or operation (class II-V) had detectable telomerase activity. In contrast, the in situ TRAP assay detected telomerase-positive cells in 85% patients before treatment and 60% patients after/during treatment. Of note is a dissociation of the results between fluorescent-based TRAP assay and that of in situ TRAP assay in some patients. Some patients without detectable telomerase activity using the fluorescent-based TRAP assay showed a low frequency of telomerase-positive cells in the urine. A combination of semiquantitative analysis and an in situ TRAP assay to detect telomerase-positive cells might be a useful tool in the detection and monitoring of patients with urological neoplasias.
Ohyashiki
K
K
First Department of Internal Medicine, Tokyo Medical College.
Yahata
N
N
Iwama
H
H
Aizawa
T
T
Miki
M
M
D006801:Humans; D012091:Repetitive Sequences, Nucleic Acid; D019098:Telomerase / Q000032:analysis*; D001749:Urinary Bladder Neoplasms / Q000175:diagnosis* / Q000628:therapy; D014556:Urine / Q000166:cytology*
D019098:Telomerase