Akademik Veri Yönetim Sistemi
Tezin Türü: Doktora
Tezin Yürütüldüğü Kurum: Bursa Uludağ Üniversitesi, FEN-EDEBİYAT FAKÜLTESİ, BİYOLOJİ, Türkiye
Tezin Onay Tarihi: 2017
Tezin Dili: Türkçe
Öğrenci: ŞEYMA AYDINLIK
Danışman: Egemen Dere
Özet:
Metastatic colon cancer has a survival rate less than %5 despite the use of effective chemotherapeutic regimen. Prognostic and predictive importance of epidermal growth factor receptor (EGFR) is still contradictive in colon cancer. For this reason, the tyrosine kinase inhibitor canertinib was used in this study to block the EGFR receptor. The effect of Palladium (Pd) (II) compound [PdCl(terpy)](sac).2H2O] which was synthesized by Uludağ University chemistry department and canertinib alone and the effect of Pd(II) compound and canertinib combination treatment was investigated in this study. 5-fluorouracil (5-FU), a chemotherapeutic drug commonly used in the treatment of cancer and its combination with canertinib was used as positive control. Therefore, the cytotoxic effects of the combination of Palladium (II) compound and EGFR inhibitor canertinib and the chemotherapeutic agent 5-fluorouracil and canertinib combination on human colon cancer cell line (HCT-15, HT-29) were also determined by the SRB viability test. Cell death (apoptosis/autophagy) responsible for the cytotoxic effects of the combination therapy, presence of apoptosis in cells and the presence of acidic vesiculations, mitochondrial depolarization were determined by flow cytometry, Annexin-V-Cy3/SYTOX dual staining method, acridine staining and tetraethylbenzimidazolylcarbocyanine iodide (JC-1) staining respectively. Alterations in proteins related with EGFR-associated survival pathway and alterations in proteins associated with epidermal mesenchymal transformation (EMT) and transformation in apoptosis/autophagy-related proteins were evaluated by western blot. In addition, effects on drug carrier proteins were first determined by SRB method using MDCKII polarize cell lines and then changes in proteins were determined by western blot. Also, effects of inhibition of this life pathway with combination therapy together on invasion, wound healing and colony forming ability were determined. Finally, effects of EGFR inhibition with combination treatments together on the ability to form tubes in HUVEC endothelial cells was assessed by in vitro matrigel test and the effects on vessel formation were also assessed by in vivo Chorio Allantoic Membrane (CAM) test. In conclusion, the combination of Pd (II) compound and canertinib has been shown to increase cytotoxic activity, induced apoptosis, decreased invasion, wound healing abilities and vessel formation in colon cancer cell lines.
Keywords: Tyrosine kinase inhibitors, Angiogenesis, Autophagy, EMT, Drug resistance