Quantitative measurement of telomerase reverse transcriptase (hTERT) mRNA in urothelial cell carcinomas

Kok, Jacques B. de; Schalken, Jack A.; Aalders, Tilly; Ruers, Theo J.M.; Willems, Hans L.; Swinkels, Dorine W.

doi:10.1002/1097-0215(20000715)87:2<217::aid-ijc10>3.0.co;2-2

Cited by 63 publications

(16 citation statements)

References 24 publications

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“…De Kok et al . studied telomerase reverse transcriptase (hTERT) expression, which closely related to telomerase activity in urothelial cell carcinomas using a real‐time quantitative PCR assay 24 . They demonstrated that an increase in both pathological grade and clinical stage as prognostic parameters correlated with increased hTERT expression.…”

Section: Discussionmentioning

confidence: 99%

Telomerase activity is correlated with lower grade and lower stage bladder carcinomas

Okumura¹,

Mizuno²,

Nagakawa³

et al. 2004

Int J of Urology

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Background : Telomerase is a ribonucleoprotein enzyme that compensates for the progressive erosion of telomeres. The increasing interest in telomerase is motivated by the demonstration that most human carcinomas are telomerase positive. The potential use of telomerase activity in bladder carcinomas using a urine sample has been reported in several studies. However, little is known about the detection of telomerase activity in bladder carcinoma tissues. Herein, we investigate telomerase activity in bladder carcinoma tissues according to grade (G) and stage. Material and methods : Telomerase activity was assayed by polymerase chain reaction enzymelinked immunosorbent assay methods. Malignant lesions were assessed in 37 patients with bladder carcinoma and no malignant lesions were assessed in five patients with dysplasia or inflammatory bladder lesions. Results : Twenty-three out of 37 carcinoma samples were telomerase-positive and one out of five control samples without carcinoma was telomerase-positive. The positive rates according to stage and grade were 83.3% for superficial and 42.1% for invasive stages and 83.3% for G1, 66.7% for G2 and 40.0% for G3. Telomerase activity was correlated with lower grade and lower stage bladder carcinomas. Conclusion : These results strongly suggest that reactivation of telomerase may differ between superficial and invasive bladder carcinomas and also between low grade and high grade bladder carcinomas.

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Section: Discussionmentioning

confidence: 99%

Telomerase activity is correlated with lower grade and lower stage bladder carcinomas

Okumura¹,

Mizuno²,

Nagakawa³

et al. 2004

Int J of Urology

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“…Total RNA was isolated from tissues and reverse transcribed to cDNA as described previously. 16 Housekeeping Genes A total of 13 housekeeping genes, used in many studies, were selected for gene expression analysis ( Table 1). All genes are constitutively expressed in various tissues.…”

Section: Tissuesmentioning

confidence: 99%

Normalization of gene expression measurements in tumor tissues: comparison of 13 endogenous control genes

Kok

Roelofs

Giesendorf

et al. 2005

Laboratory Investigation

494

332

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For interpretation of quantitative gene expression measurements in clinical tumor samples, a normalizer is necessary to correct expression data for differences in cellular input, RNA quality, and RT efficiency between samples. In many studies, a single housekeeping gene is used for normalization. However, no unequivocal single reference gene (with proven invariable expression between cells) has been identified yet. As the best alternative, the mean expression of multiple housekeeping genes can be used for normalization. In this study, no attempt was made to determine the gold-standard gene for normalization, but to identify the best single housekeeping gene that could accurately replace the measurement of multiple genes. Expression patterns of 13 frequently used housekeeping genes were determined in 80 normal and tumor samples from colorectal, breast, prostate, skin, and bladder tissues with real-time quantitative RT-PCR. These genes included, large ribosomal protein, b-actin, cyclophilin A, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerokinase 1, b-2-microglobin, b-glucuronidase, hypoxanthine ribosyltransferase (HPRT), TATA-box-binding protein, transferrin receptor, porphobilinogen deaminase, ATP synthase 6, and 18S ribosomal RNA. Principal component analysis was used to analyze these expression patterns, independent of the level of expression. Our approach identified HPRT as the single best reference gene that could be used as an accurate and economic alternative for the measurement of multiple housekeeping genes. We recommend this gene for future studies to standardize gene expression measurements in cancer research and tumor diagnostics until a definite gold standard has been determined.

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“…This has been con®rmed for the speci®c case of tissue samples obtained from carcinomas of the urinary bladder (Ito et al, 1998a). In addition, using a quantitative, realtime PCR method, it could be shown that hTERT levels also correlate with both the tumour stage and its degree of di erentiation (de Kok et al, 2000b).…”

Section: Rt ± Pcr Detection Of Htert In the Diagnosis Of Carcinoma Ofmentioning

confidence: 99%

Telomerase: Its clinical relevance in the diagnosis of bladder cancer

Müller

2002

Oncogene

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More than 50 years ago, Papanicolaou recognized the importance of a non-invasive technique for the diagnosis and follow-up of patients with carcinoma of the urinary bladder. Cystoscopy, however, has remained the`gold standard' since no currently available non-invasive method can compete with cystoscopy's sensitivity and speci®city. The detection of the ribonucleoprotein telomerase or the telomerase subunits human telomerase RNA (hTR) and human telomerase reverse transcriptase (hTERT) in urine samples o er new diagnostic perspectives. The present article presents a review of publications in the literature and evaluates their clinical relevance. The experimental studies reported to date are very promising and show that telomerase exactly ful®ls the requirements for a good diagnostic marker for carcinoma of the urinary bladder. The diagnostic application remains in an experimental stage and telomerase is still several steps away for routine use as a clinical parameter. The remaining steps leading to its routine clinical application will be discussed.

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Quantitative measurement of telomerase reverse transcriptase (hTERT) mRNA in urothelial cell carcinomas

Cited by 63 publications

References 24 publications

Telomerase activity is correlated with lower grade and lower stage bladder carcinomas

Telomerase activity is correlated with lower grade and lower stage bladder carcinomas

Normalization of gene expression measurements in tumor tissues: comparison of 13 endogenous control genes

Telomerase: Its clinical relevance in the diagnosis of bladder cancer

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