Mikrotübül İnhibitörlerinin Kanser Hücrelerinde Tehlike-İlişkili Moleküler Desen Düzeylerine Etkisinin Araştırılması

Abstract

Cancer cells are proliferate rapidly and uncontrolled. Chemotherapeutics such as vinorelbine and colchicine affect the cell cycle and DNA synthesis mechanism; they causes cells to be kept in the G2 / M phase by preventing the polymerization or depolymerization of microtubules. Vinorelbine binds to the vinca binding site at the end of the microtubules, preventing the elongation of microtubules. Colchicine forms a tubulin-colchicine complex by binding to tubulin dimers in soluble form. Within the scope of the hypothesis of this study, the increase of immunological danger-associated molecular pattern (DAMP) molecules in control fibroblasts and cancer cells arrested in the G2 / M phase due to microtubule inhibitors applied at a concentration that does not affect cell viability was investigated. For this purpose, the subtoxic concentration of vinorelbine and colchicine in NIH / 3T3, L929, 4T1, B16-F10 cells was determined. With the application of these agents for 10 hours and 20 hours, an increase in the G2 / M ratio was observed. When serum starvation was applied in order to synchronize these cells, G0 / G1 phase arrest was observed. When vinorelbine and colchicine were applied to synchronized cells, a significant increase in G2 / M ratio was observed in 20 hours of incubation. These findings were confirmed by imaging the disruption in the cytoskeleton. In order to determine the increase in DAMP caused by cellular stress developing under cell cycle blockage, the change in the expression levels of 7 different DAMP genes was examined. An increase in mRNA level of Hmgb1, Nlrp3, Il33, Hsp70, Hsp90, Ppia and H3.3 DAMP genes was determined with the effect of vinorelbine and colchicine. The stress situation was also confirmed by the increase of DNA molecule released outside the cell and the translocation of the HMGB1 protein from nucleus to cytoplasm. Consequently, vinorelbine and colchicine disrupted microtubule polymerization, causing cell cycle arrest in G2 / M phase. There is new evidence that this blockade in the cell cycle creates cellular stress and can lead to an increase in DAMP expression.