Comparison of the telomeric repeat amplification protocol (TRAP) to the new TRAP-eze telomerase detection kit
Tóm tắt
The ribonucleoprotein, telomerase, is believed to be responsible for the maintenance of telomere length in immortal and cancer cells. A PCR-based assay for the detection of telomerase activity (TRAP assay: telomeric repeat amplification protocol) was developed, allowing fast and efficient detection of telomerase activity when sample amounts are limiting. Of the thousands of primary human tumors examined using the TRAP assay, almost 90% have been shown to exhibit telomerase activity. Thus, for the early detection of cancer and for the rapid screening of compounds and drugs in cancer therapeutics, methods for the detection of telomerase activity are rapidly emerging. The recently developed TRAP-ezeTM kit from Oncor, Inc. gives increased sensitivity with decrease sample processing time, allowing improved detection of telomerase activity in a large number of samples. In the present study, we have addressed some of the technical aspects and limitations of critical importance for reproducibility, reliability, and linearity of the standard TRAP assay and the TRAP-ezeTM kit using cell culture and clinical materials.
Tài liệu tham khảo
Allsopp RC, Vaziri H, Patterson C, Goldstein S, Younglai EV, Futcher AB, Greider CW, Harley CB (1992). Telomere length predicts replicative capacity of human fibroblasts. Proc Natl Acad Sci (USA) 89: 10114–10118.
Broccolli D, Young JW, de Lange T (1995). Telomerase activity in normal and malignant hematopoietic cells. Proc Natl Acad Sci (USA) 92: 9082–9086.
Counter CM, Avillon AA, LeFeuvre CE, Stewart NG, Greider CW, Harley CB, Bacchetti S (1992). Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity. EMBO J 11: 1921–1929.
Counter CM, Botelho FM, Wang P, Harley CB, Bacchetti S (1994). Stabilization of short telomeres and telomerase activity accompany immortalization of Epstein-Barr virus-transformed human B lymphocytes. J Virol 68: 3410–3414.
Counter CM, Hirte HW, Bacchetti S, Harley CB (1994). Telomerase activity in human ovarian carcinoma. Proc Natl Acad Sci (USA) 91: 2900–2904.
Greider CW, Blackburn EH (1985). Identification of a specific telomere terminal transferase activity inTetrahymena extracts. Cell 43: 405–413.
Greider CW, Blackburn EH (1989). A telomeric sequence in the RNA ofTetrahymena telomerase required for telomere repeat synthesis. Nature 337: 331–337.
Harley CB, Futcher AB, Greider CW (1990). Telomeres shorten during aging. Nature 345: 458–460.
Harris CC (1987). Human tissues and cells in carcinogenesis research. Cancer Res 47: 1–10.
Hastie ND, Dempster M, Dunlop MG, Thompson AM, Green DK, Allshire RC (1990). Telomere reduction in human colorectal carcinoma and with ageing. Nature 346: 866–868.
Hiyama E, Hiyama K, Yokoyama T, Matsuura Y, Piatyszek MA, Shay JW (1995). Correlation of telomerase activity level with human neuroblastoma outcomes. Nature Medicine 1: 249–257.
Hiyama K, Hiyama E, Ishoika S, Yamakido M, Inai K, Gazdar AF, Piatyszek MA, Shay JW (1995). Telomerase activity in small-cell and non-small-cell lung cancers. J Natl Cancer Inst 87: 895–902.
Hiyama K, Hirai Y, Kyoizumi S, Akiyama M, Hiyama E, Piatyszek MA, Shay JW, Ishoika S, Yamakido M (1995). Activation of telomerase in human lymphocytes and hematopoietic progenitor cells. J Immuno 155: 3711–3715.
Hiyama E, Yokoyama T, Tatsumoto N, Hiyama K, Imamura Y, Murakami Y, Kodama T, Piatyszek M, Shay JW, Matsuura Y (1995). Telomerase activity in gastric cancer. Cancer Res 55: 3258–3262.
Hiyama E, Gollahon L, Kataoka T, Kuroi K, Yokoyama T, Gazdar AF, Hiyama K, Piatyszek MA, Shay JW (1996). Telomerase activity in human breast tumors. J Natl Cancer Inst 88: 116–122.
Holt SE, Wright WE, Shay JW (1996). Regulation of telomerase activity in immortal cell lines. Molec Cell Biol 16: 2932–2939.
Holt SE, Shay JW, Wright WE (1996). Refining the Telomere-Telomerase hypothesis of aging and cancer. Nature Biotech 14: 834–837.
Huschtscha LL, Holliday R (1983). Limited and unlimited growth of SV40-transformed cells from human diploid MRC-5 fibroblasts. J Cell Sci 63: 77–99.
Kaczorowski T, Furmanek B, Sektas MA (1994). Method for removal of radioactive nucleotides from electrophoretic buffers. BioTechniques 1994; 16: 1030–1031.
Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PLC, Coviello GM, Wright WE, Weinrich SL, Shay JW (1994). Specific association of human telomerase activity with immortal cells and cancer. Science 266: 2011–2015.
Levy MZ, Allsopp RC, Futcher AB, Greider CW, Harley CB (1992). Telomere end-replication problem and cell aging. J Mol Biol 225: 951–960.
Morin GB (1989). The human telomere terminal transferase enzyme is a ribonucleoprotein that synthesizes TTAGGG repeats. Cell 59: 521–529.
Norton JC, Piatyszek MA, Wright WE, Shay JW, Corey DR (1996). Inhibition of human telomerase activity by peptide nucleic acids. Nature Biotech 14: 1615–1619.
Olovnikov AM (1973). A theory of marginotomy. J Theor Biol 41: 181–190.
Piatyszek MA, Kim NW, Weinrich SL, Hiyama K, Hiyama E, Wright WE, Shay JW (1995). Detection of telomerase activity in human cells and tumors by a telomeric repeat amplification protocol (TRAP). Meth Cell Sci 17: 1–15.
Sharma HW, Sokoloski JA, Perez JR, Maltese JY, Sartorelli AC, Stein CA, Nichols G, Khaled Z, Telang NT, Narayaman R (1995). Differentiation of immortal cells inhibits telomerase activity. Proc Natl Acad Sci USA 92: 12343–12346.
Shay JW, Pereira-Smith OM, Wright WE (1991). A role for both Rb and p53 in the regulation of human cellular senescence. Exp Cell Res 196: 33–39.
Shay JW, Wright WE, Werbin H (1991). Defining the molecular mechanisms of human cell immortalization. Biochem Biophys Acta 1072: 1–7.
Sommerfeld HJ, Meeker AK, Piatyszek MA, Bova GS, Shay JW, Coffey DS (1995). Telomerase activity: A prevalent marker of malignant human prostate tissue. Cancer Res 56: 218–222.
Taylor RS, Ramirez RD, Ogoshi M, Chaffins M, Piatyszek MA, Shay JW (1996). Detection of telomerase activity in malignant and nonmalignant skin conditions. J Invest Dermatol 106: 759–765.
Vaziri H, Schächter F, Uchida I, Wei L, Zhu X, Effros R, Cohen D, Harley CB (1993). Loss of telomeric DNA during aging of normal and trisomy 21 human lymphocytes. Am J Hum Genet 52: 661–667.
Watson JD (1972). Origin of concatameric T4 DNA. Nature 239: 197–201.
Wright WE, Pereira-Smith OM, Shay JW (1989). Reversible cellular senescence: Implications for a two-stage model for the immortalization of normal human diploid fibroblasts. Mol Cell Biol 9: 3088–3092.
Wright WE, Shay JW, Piatyszek MA (1995). Modifications of a telomeric repeat amplification protocol (TRAP) result in increased reliability, linearity and sensitivity. Nuc Acids Res 23: 3794–3795.
Wright WE, Shay JW (1992). The two-stage mechanism controlling cellular senescence and immortalization. Exp Geron 27: 383–389.
Wright WE, Shay JW (1992). Telomere positional effects and the regulation of cellular senescence. Trends Genet 8: 193–197.