| dc.description.abstract | This bachelor thesis focuses on the effect of mitochondrial inhibitors CCCP, rotenone and oligomycin on the two different pancreatic cancer cell lines PANC-1 and BxPC-3. Cell viability was tested after the cells were exposed for 24 and 48 hours to different concentrations of the inhibitors. To gain a better understanding of the metabolic characteristics of PANC-1 and BxPC-3 after treatment, the cells were also tested for oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) using the Agilent Seahorse XFe96 Analyzer. To account for difference in sample loading and cell number, a BCA protein assay was carried out after the Seahorse assay, to normalize the measures from Seahorse to the corresponding protein concentrations.
The results showed that PANC-1 were more dependent on oxidative phosphorylation as fluorescence values decreased with increasing concentration, meaning the inhibitors displayed an effect on this cell line. However, exposure time did not show any consistent trend. The 48-hour treatment generally exhibited greater fluorescence compared to the 24-hour treatment, possibly due to compensatory metabolic changes. From the Seahorse energy map, oligomycin treatment exhibited the most substantial effect.
The BxPC-3 cancer cells were found to be more glycolytic, as possible compensatory mechanisms whereby cell viability increased with increasing drug concentration. Based on the results obtained from the Seahorse Analyzer, neither of the drugs caused a large shift in metabolism in this cell line.
Overall, these experiments demonstrated how cancer cell lines respond differently to the same treatments. The results exhibited some large standard deviations, suggesting that there was a substantial amount of variability in the data which needs further investigating. Therefore, it is important to perform additional experimentation to confirm the findings and obtain more measurements. | |