TRAP-silver staining, a highly sensitive assay for measuring telomerase activity in tumor tissue and cell lines

• Autor(a) principal: Dalla Torre, Cristiane Arruda [UNIFESP]
• Data de Publicação: 2002
• Outros Autores: Maciel, Rui Monteiro de Barros [UNIFESP], Pinheiro, Nídia Alice [UNIFESP], Andrade, Joyce Anderson Duffles [UNIFESP], Toledo, Silvia Regina Caminada de [UNIFESP], Villa, Luisa Lina, Cerutti, Janete Maria [UNIFESP]
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UNIFESP
• Texto Completo: http://dx.doi.org/10.1590/S0100-879X2002000100009; http://repositorio.unifesp.br/handle/11600/1315
• topic: Telomeric repeat amplification protocol; TRAP assay; Human thyroid cancer; Leukemia; Telomerase; Silver staining

Measurement of telomerase activity in clinically obtained tumor samples may provide important information for use as both a diagnostic marker and a prognostic indicator for patient outcome. In order to evaluate telomerase activity in tumor tissue without radiolabeling the product, we developed a simple telomeric repeat amplification protocol-silver-staining assay that is less time-consuming, is safe and requires minimal equipment. In addition, we determined the sensitivity of the silver-staining method by using extracts of telomerase-positive thyroid carcinoma cell lines which were serially diluted from 5,000 to 10 cells. Telomerase activity was also assayed in 19 thyroid tumors, 2 normal controls and 27 bone marrow aspirates. The results indicate that the technique permits the detection of telomerase activity from 5000 to as few as 10 cells. We propose that it could be immediately applicable in many laboratories due to the minimal amount of equipment required.