Müller M, Krause H, Heicappell R, Tischendorf J, Shay J W, Miller K
Department of Urology, Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, Germany.
Clin Cancer Res. 1998 Aug;4(8):1949-54.
New, noninvasive methods for the early detection of urothelial carcinomas of the urinary bladder are needed for the diagnosis, follow-up, and screening of patients with bladder cancer. Detection of the enzyme telomerase in urine could offer these new diagnostic possibilities. The standard technique for detecting telomerase activity is the telomeric repeat amplification protocol (TRAP assay). Because of the instability of the ribonucleoprotein telomerase in an aggressive medium, such as urine, investigations conducted to date have yielded nonuniform or even contradictory findings. This study compares the detection of human telomerase RNA (hTR) by reverse transcriptase-PCR (RT-PCR) with detection of telomerase activity by the TRAP assay in the diagnosis of urothelial carcinoma of the urinary bladder. Sedimented cells obtained from urine of 30 patients with urothelial carcinoma, 15 patients with benign urological disorders, 3 patients as part of follow-up for malignant disease, and 20 healthy subjects were examined for the presence of hTR and for telomerase activity (TRAP). In patients with bladder cancer, telomerase activity was detected by the TRAP assay in only 2 of 30 specimens (7%). However, increased levels of hTR were detected by RT-PCR in 25 of the same 30 cases (83%). For patients with benign urological disorders, such as urolithiasis or urinary tract infections, hTR was detected in samples obtained from 4 of 15 patients (27%). Low hTR expression levels were found in 15% of the healthy controls. The detection of hTR by RT-PCR represents a promising new method for detecting malignant cells in urine.
膀胱癌的诊断、随访及筛查需要新的非侵入性方法来早期检测膀胱尿路上皮癌。检测尿液中的端粒酶可为这些新的诊断提供可能。检测端粒酶活性的标准技术是端粒重复序列扩增法（TRAP 检测）。由于核糖核蛋白端粒酶在尿液等活性介质中不稳定，迄今为止进行的研究结果并不一致甚至相互矛盾。本研究比较了逆转录聚合酶链反应（RT-PCR）检测人端粒酶 RNA（hTR）与 TRAP 检测端粒酶活性在膀胱尿路上皮癌诊断中的应用。对 30 例膀胱尿路上皮癌患者、15 例良性泌尿系统疾病患者、3 例恶性疾病随访患者以及 20 名健康受试者尿液中的沉淀细胞进行 hTR 检测及端粒酶活性（TRAP）检测。在膀胱癌患者中，TRAP 检测仅在 30 份标本中的 2 份（7%）检测到端粒酶活性。然而，RT-PCR 在相同的 30 例患者中的 25 例（83%）检测到 hTR 水平升高。对于患有良性泌尿系统疾病（如尿路结石或尿路感染）的患者，15 例患者中有 4 例（27%）的样本检测到 hTR。15%的健康对照者中发现 hTR 表达水平较低。RT-PCR 检测 hTR 是一种很有前景的检测尿液中恶性细胞的新方法。
