| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Enzymes and Protein Markers |
1
Clinical Biochemistry Unit, Department of Clinical Physiopathology, University of Florence, 50139 Florence, Italy.
2
Clinical Laboratory Department, Azienda Ospedaliera
Careggi, 50139 Florence, Italy.
3
Endocrinology Unit, Department of Clinical
Physiopathology, University of Florence, 50139 Florence, Italy.
4
Institute of General Pathology, University of Florence,
50139 Florence, Italy.
a Address correspondence to this author at: Clinical Biochemistry Unit, Department of Clinical Physiopathology, viale Pieraccini 6, 50139 Florence, Italy. Fax 39-55-4377290; e-mail c.orlando{at}dfc.unifi.it.
Telomerase is a ribonucleoprotein enzyme that adds TTAGGG repeats onto human telomeres, preventing their shortening. The activation of this enzyme is an important step in cell immortalization and carcinogenesis and seems to represent a new and promising marker in cancer diagnosis and management. Telomerase activity is usually detected in cellular protein extract by the telomeric repeat amplification protocol (TRAP) assay, which can provide only a qualitative (presence/absence) evaluation. Here we present a modification of this method that can provide quantitative information without requiring time-consuming post-PCR procedures such as gel electrophoresis with radioactive materials and autoradiography. The detection and measurement of telomerase activity is performed by evaluating the amount of double-stranded DNA generated in the telomerase reaction and PCR amplification, with the use of the sensitive DNA fluorescent dye PicoGreen®. In a subset of tumors, the presence of telomerase activity was confirmed by the conventional TRAP assay. By this method we evaluated telomerase activity in unselected groups of breast (n = 15), ovarian (n = 12), endometrial (n = 12), gastric (n = 20), and renal (n = 12) carcinomas, in meningiomas (n = 8), and in pheochromocitomas (n = 10). The results indicate substantial differences of telomerase activity among cancer groups; however, a large variability among patients of the same group is observed. Kidney, ovarian, and breast carcinomas showed the highest mean values (31.8 ± 28.9, 29.2 ± 26.7, and 35.3 ± 15.9 ng DNA/µg protein, respectively, mean ± SD), whereas gastric and endometrial cancers had a lower activity (17.2 ± 8.8 and 13.5 ± 7.9 ng DNA/µg protein, respectively). Very low or no detectable telomerase activity was found in meningiomas (with the exception of one malignant atypical variant) and pheochromocitomas (9.7 ± 12.9 and 2.8 ± 2.1 ng DNA/µg protein, respectively). In conclusion, our method seems to be an accurate and reasonable procedure for measuring telomerase activity in human cancers.
The following articles in journals at HighWire Press have cited this article:
|
|
 |
|
  K. Ozturk, M. Durusoy, and E. Piskin A Simple Quartz Crystal Microbalance Nucleic Acid Sensor for Detection of Telomerase Journal of Bioactive and Compatible Polymers, November 1, 2008; 23(6): 579 - 593. [Abstract] [PDF]
|
|
|
|
 |
|
 J. Xing, Y. Zhu, H. Zhao, H. Yang, M. Chen, M. R. Spitz, and X. Wu Differential Induction in Telomerase Activity among Bladder Cancer Patients and Controls on {gamma}-Radiation Cancer Epidemiol. Biomarkers Prev., March 1, 2007; 16(3): 606 - 609. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Sanchini, R. Gunelli, O. Nanni, S. Bravaccini, C. Fabbri, A. Sermasi, E. Bercovich, A. Ravaioli, D. Amadori, and D. Calistri Relevance of Urine Telomerase in the Diagnosis of Bladder Cancer JAMA, October 26, 2005; 294(16): 2052 - 2056. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Grimm, J M. Perez, L. Josephson, and R. Weissleder Novel Nanosensors for Rapid Analysis of Telomerase Activity Cancer Res., January 15, 2004; 64(2): 639 - 643. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Wege, M. S. Chui, H. T. Le, J. M. Tran, and M. A. Zern SYBR Green real-time telomeric repeat amplification protocol for the rapid quantification of telomerase activity Nucleic Acids Res., January 15, 2002; 31(2): e3 - e3. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Peri, P. Luciani, B. Conforti, S. Baglioni-Peri, F. Cioppi, C. Crescioli, P. Ferruzzi, S. Gelmini, G. Arnaldi, G. Nesi, et al. Variable Expression of the Transcription Factors cAMP Response Element-Binding Protein and Inducible cAMP Early Repressor in the Normal Adrenal Cortex and in Adrenocortical Adenomas and Carcinomas J. Clin. Endocrinol. Metab., November 1, 2001; 86(11): 5443 - 5449. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Gelmini, A. Crisci, B. Salvadori, M. Pazzagli, C. Selli, and C. Orlando Comparison of Telomerase Activity in Bladder Carcinoma and Exfoliated Cells Collected in Urine and Bladder Washings, Using a Quantitative Assay Clin. Cancer Res., July 1, 2000; 6(7): 2771 - 2776. [Abstract] [Full Text]
|
|
|
|
|
|
M. Mannelli, S. Gelmini, G. Arnaldi, L. Becherini, D. Bemporad, C. Crescioli, M. Pazzagli, F. Mantero, M. Serio, and C. Orlando Telomerase Activity Is Significantly Enhanced in Malignant Adrenocortical Tumors in Comparison to Benign Adrenocortical Adenomas J. Clin. Endocrinol. Metab., January 1, 2000; 85(1): 468 - 470. [Abstract] [Full Text]
|
|
|
|
|
|
P. Tangkijvanich, D. Tresukosol, P. Sampatanukul, S. Sakdikul, N. Voravud, V. Mahachai, and A. Mutirangura Telomerase Assay for Differentiating between Malignancy-related and Nonmalignant Ascites Clin. Cancer Res., September 1, 1999; 5(9): 2470 - 2475. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
