Optimized Detection Method of Telomerase Activity in Cancer Diagnostics

摘要

Telomerase is a ribonucleoprotein complex; it uses an internal RNA template to synthesize telomere DNA. Telomerase is active in 90% of cancers and can be used as a diagnostic marker. We have optimized conditions for the extraction from small tissue samples (<0.05 g) of cervical lesions to analyze telomerase activity and selected the optimal concentrations of the tissue extracts. Different concentrations of the extracts were used to determine the presence of possible telomerase inhibitors and Taq-polymerase in the extracts. Using lung and kidney cancer samples it was shown that these conditions are applicable for the estimation of telomerase activity in different cancer types. Many investigations of telomerase activity using different types of TRAP (Telomere Repeat Amplification Protocol) have been performed. The possibility of comparison of TRAP results with radioactive and Sybr green detection remains open. We compared these two types of detection for several samples of cervical intraepithelial neoplasias and conclude that they have similar sensitivities.