Quantitative evaluation of telomerase subunits in urine as biomarkers for noninvasive detection of bladder cancer

Abstract

The aim of our study was to prospectively evaluate the potential diagnostic value and clinical applicability of quantitative analysis of telomerase subunits gene expression in urine for noninvasive detection of bladder cancer. Expression levels of human telomerase reverse transcriptase (hTERT) and human telomerase RNA (hTR) were analyzed by real-time reverse transcriptase polymerase chain reaction (RT-PCR) in urine samples from 163 subjects with bladder cancer and 237 controls (163 individuals with benign genitourinary diseases; 74 healthy subjects). The sensitivity, specificity and optimal cutoffs were determined and compared to the corresponding values obtained by voided urine cytology. Quantitative urinary hTR analysis detects bladder cancer with an overall sensitivity of 77.0%, whereas hTERT analysis reached a sensitivity of 55.2%. The majority of undetected tumors were small, lowgrade pTa lesions. Both hTR and hTERT proved to be significantly more sensitive than cytology (34.5%; p < 0.001). Specificities for hTR, hTERT and cytology were 72.1%, 85.0% and 92.7%, respectively, in the total study population and 96.9%, 89.2% and 100%, respectively, in healthy subjects. Higher diagnostic accuracy was achieved by hTR than by hTERT analysis (p < 0.05). The specificity of hTR increased to 85.0% in the total population if urinary leukocyte contamination was excluded. These data suggest that quantitative hTR analysis is the most accurate telomerase-based test for bladder cancer detection and has the potential to replace cytology as a noninvasive biomarker for disease diagnosis and follow-up. ' 2005 Wiley-Liss, Inc.

Detailed Technical Analysis of Urine RNA-Based Tumor Diagnostics Reveals ETS2/Urokinase Plasminogen Activator to Be a Novel Marker for Bladder Cancer

“…Stratification by tumor grade (Fig. 3A) indicated a higher sensitivity for high-grade tumors (81.3%, grade 2-3), an observation that has been described for other urine-based RNA markers (14,23,24 ).…”

“…The feasibility of the diagnostic application has been demonstrated by prospective studies (14 ), but different authors report substantially different diagnostic accuracies, even if the same markers were used. Using the monovette-based sampling of whole urine presented here provides several technical advantages.…”

“…RNA extracts from tumor material (8)(9)(10), bladder washings (11,12 ), and exfoliated cells (13 ) have been used for RT-qPCR-based hTERT mRNA quantification for the detection of bladder cancer, resulting in reported diagnostic sensitivities of 50% to 100%. When urine sediment was used as sample material in a larger prospective study, however, hTERT mRNA expression was often found to be near the detection limit, resulting in a sensitivity of 55% (14 ).…”

TERT Promoter Mutations and TERT mRNA but Not FGFR3 Mutations Are Urinary Biomarkers in Han Chinese Patients With Urothelial Bladder Cancer

“…Consequently, telomerase silence and activation in normal and malignant cells results generally from the repression and derepression, respectively, of the TERT gene. Numerous studies have evaluated telomerase activity and TERT as cancer diagnostic markers and therapeutic targets, given their cancer-specific expression or activation [3,[5][6][7].Widespread telomerase activation and TERT expression, for example, occur in UBC, and detection of telomerase activity and/or TERT mRNA in voided urine has been successfully applied to urine-based diagnostics for and recurrence surveillance of UBC [8][9][10][11][12][13][14][15][16][17][18]; however, non-tumorderived TERT expression poses a potential drawback. Inflammatory lymphocytes, for example, may contain substantial amounts of TERT transcripts or telomerase activity and may cause a false-positive result when exfoliated into urine [8].…”

Development of noninvasive bladder cancer diagnosis on basis of telomerase and it’s subunits hTERT and hTR detection

“…необходимые для прогнозирования течения болезни и последующего врачебного наблюдения за пациентами [5,6]. Временной фактор является важным для диагностики и успешности лечения РМП, поэтому выявление онкомаркёров на ранней стадии заболевания является актуальной задачей современной онкологии [6].…”