Аннотация:Telomerase is a ribonucleoprotein complex, which copy an internal RNA template to synthesize telomere DNA, which are shortened during cell divisions. Telomerase is active in about 80-90% of cancers, but not in most of somatic tissues. We optimized conditions for extraction from small tissue samples (<0,05 g) and selected optimal concentrations of tissue extract concentration for cervical lesions. Two concentrations of extracts in reaction were used to take into account in-sample inhibitors. We use these conditions for several cancerous and normal samples of lung and kidney, these conditions are identical for different types of cancer. There are a lot of investigations of telomerase activity using different types of TRAP (Telomere Repeat Amplification Protocol). We compare two types of detection (radioactive and Sybr green) on the several samples of cervical intraepithelial neoplasias and conclude, that sensibility is equal. Telomerase activity resulting in lengthening of telomeres and increase of generation’s number in epithelial cells is found in all tumors. The simplified variant of widely known test of telomerase activity has been developed for clinical samples. To solve the problem of using telomerase activity as diagnostic marker telomerase activation was studied in a spectrum of tumors at different developmental stages and in precancerous lesions, such as cervical intraepithelial neoplasias.
It is known different classes of telomerase inhibitors. The first is nucleotide analogs. They inhibit the polymerase activity of telomerase, but they also inhibit all other cellular polymerase. The second type of inhibitors is oligonucleotides that blocked the template region of telomerase RNA. The other one is substances that destroy the quadruplex structure in telomere region.
We checked the effect on telomerase activity of oligonucleotides that interacting with telomerase RNA in the region (distinct from template region) that promote the assembly of telomerase.
The other way to decrease the telomerase activity is prevent the synthesis of telomerase component. We tried to prevent the synthesis of telomerase by transfection of siRNA in the cell culture.
Development of novel test and fundamental study of telomerase complex are other ways of project.
The developed test and fundamental research of telomerase have been used for checking classes of proposed telomerase inhibitors and for creation of new one.