Effect of resveratrol on telomerase activity due to NF-KB and hTERT expression in PANC-1 cell line
Project no. 20730 · Status: Kapanmış (Yayın bekleniyor) · 2019
Summary
Pancreatic cancer, which is quite difficult to diagnose and differentiate from other pancreatic diseases, is one of the deadliest malignant neoplasms in the world, especially in developed countries. Immortality, which is caused by the unlimited ability of proliferation in cancer cells, is associated with increased activity of the telomerase enzyme. Telomerase activation is a critical step in the development of about 85% of human cancers and it is not yet fully understood how telomerase is reactivated in cancer cells. Therefore, inhibition of telomerase activity in cancer treatment seems to be one of the important treatment options. In this thesis, the effects of Resveratrol, a natural compound in pancreatic ductal adenocarcinoma cell line, PANC-1, on genes associated with telomerase, on hTERT and NF-?B gene expression and thus on pancreatic cancer development. Proliferation and cytotoxicity analyzes were carried out with the WST-1 reagent by replicating the PANC-1 cell line under the appropriate environment and conditions and IC50 value for Resveratrol was determined in the light of these. By using the determined dose of IC50; The expression levels of hTERT and NF-?B, which we determined as apoptosis, wound closure analysis and telomerase related genes, were evaluated by RT-PCR analysis. In the light of the findings we found, Resveratrol has been found to have a cytotoxic effect on the PANC-1 cell line, and its IC50 value was calculated as 75.80 µM at 72 hours. The apoptotic effect was evaluated by applying the determined IC50 value to the PANC-1 cell line. Apoptosis results were evaluated with Annexin V and according to the results, 41.9% apoptotic effect was detected. When the effect of resveratol on cell migration and invasion in the PANC-1 cell line was again applied by applying IC50 value, it was found that it caused a delay in closure of the wound line at the 72nd hour compared to the control, preventing cell migration. hTERT and NF-?B gene expressions, the genes we detected in relation to telomerase, were converted to cDNA by using IC50 dosed PANC-1 cell line and evaluated by RT- PCR in LightCycler 480 device. As a result, Resveratrol has been found to cause an increase in the expression level of 1.37 times in the hTERT gene and 1.55 times in the NF-?B gene. As a result, Resveratrol showed cytotoxic effect on the PANC-1 cell line, causing apoptosis. It also prevented cell migration by preventing cell migration. However, the gene expression we associate with telomerase did not decrease as we wanted. Therefore, our study requires continuity by expanding the scope of the experiment. Keywords; Resveratrol; telomerase; hTERT; NF- KB; pancreatic cancer