Assessment of Hyperthermia and Chemotherapy Effects on Mitochondrial Respiratory Functions in Pancreatic Cancer Cells

The final master thesis prepared by Vaida Stasiulytė “Assessment of Hyperthermia and Chemotherapy Effects on Mitochondrial Respiratory Functions in Pancreatic Cancer Cells”/ Advisor Prof. Dr. S. Trumbeckaitė; Lithuanian University of Health Sciences, Faculty of Pharmacy, Department of Pharmocognosy. – Kaunas. The aim of the thesis is to measure pancreatic cancer cell (T3M4) mitochondrial respiratory rates in cases when cells are affected and not affected by chemotherapeutic drug cisplatin and/ or hyperthermia. The tasks of the thesis are the following: to measure and compare pancreatic cancer cell (T3M4) mitochondrial respiratory rates in different metabolic states in both cases when the cells are affected and are not affected by cisplatin; to measure and compare pancreatic cancer cell mitochondrial respiratory rates when cells are affected by hyperthermia (40°C and 43°C); to investigate the effect of cisplatin and hyperthermia on the permeability of mitochondrial outer and inner membrane; and to investigate the effect of chemotherapeutic hyperthermia (combination of cisplatin and 40°C, 43°C temperature) on pancreatic cancer cell (T3M4) mitochondrial respiratory rates. Methodology used to perform the tasks of the thesis is the following: the pancreatic cancer cell line (T3M4) was divided into controls and research groups affected by chemotherapeutic drug cisplatin (IC50 48µM) and/ or hyperthermia (40°C and 43°C temperature). The pancreatic cancer cell mitochondrial respiratory rates were measured by oxygraphic method (at 37°C temperature), which is performed using “Oroboros Oxygraph-2k” oxygraph. The results of the reasearch are the following: the chemotherapeutic drug cisplatin increases the permeability of mitochondrial inner membrane and inhibits ATP synthesis in pancreatic cancer cells (T3M4) with both glutamate/malate and succinate and increases the permeability of mitochondrial outer membrane. The research demonstrates that hyperthermia has no effect on pancreatic cancer cell mitochondrial respiratory rates. The chemotherapeutic hyperthermia also increases the permeability of pancreatic cancer cell mitochondrial inner membrane. Nevertheless, this effect is observed due to application of cisplatin rather than hyperthermia. The conclusions are the following: cisplatin and chemotherapeutic hyperthermia uncouples mitochondrial oxidative – phosphorylation processes of pancreatic cancer cells (T3M4) and increase the permeability of mitochondrial inner and outer membrane. Hyperthermia has no effect on pancreatic cancer cells (T3M4) and shows no additional synergistic effect on mitochondrial respiratory functions