Moloney, Andrew Hugo Dominic (2025) The characterization of the enzyme kinetics of lactate dehydrogenase and its inhibition using three novel silver organometallic compounds. Masters thesis, York St John University.
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The characterization of the enzyme kinetics of lactate dehydrogenase and its inhibition using three novel silver organometallic compounds.pdf - Published Version Available under License Creative Commons Attribution. | Preview |
Abstract
Previous research by the university of Huddersfield identified a panel of silver N-heterocyclic carbine complexes (Ag-NHCs) functioning as potential inhibitors of glycolytic lactate production. Early work explored the inhibitory potential of Ag-NHCs in various cancer cell lines. Here, this is extended by analysing the impact of Ag-NHCs on pancreatic and colorectal cancer cell viability and lactate production to ascertain their potential use in chemotherapy regimens.
A standard LDH activity assay was adapted to investigate extracellular lactate utilising the high sensitivity of NADH absorbance at 340nm. The study measured the absorbance change associated with the reduction of NAD+ to NADH during lactate conversion to pyruvate. Culture media was sampled from cancer cells grown with or without various glycolytic inhibitors (including three novel Ag-NHC compounds Ag8, HA197, and HA266). Extracellular lactate released into media was assayed in vitro following the addition of reactants (LDH and NAD+ ), where the rate of reaction is considered proportional to the concentration of extracellular lactate. This study showed all three novel Ag-NHCs reduced extracellular lactate levels to a greater extent than two established glycolysis inhibitors (silibinin and sodium oxamate). Furthermore, in all cases cell lines were most sensitive to Ag8.
The chemotherapeutic properties of the compounds were further investigated through assessment of cell viability using the resazurin metabolic activity assay. Following exposure to the novel Ag-NHCs, IC50 values were determined for various cancer cell lines and compared with normal human fibroblasts as a control for cancer cell specificity. Additionally, cells were co-treated with cisplatin and Ag-NHCs, where the efficacy of cisplatin was enhanced by the presence of Ag-NHC compounds. This study demonstrated that all cell lines were most sensitive to Ag8, which displayed the lowest IC50 values. Overall, this study established three Ag-NHCs as novel inhibitors of lactate production, apoptosis inducers, and potential chemo-sensitising agents.
| Item Type: | Thesis (Masters) |
|---|---|
| Status: | Published |
| Subjects: | Q Science > Q Science (General) |
| School/Department: | School of Science, Technology and Health |
| URI: | https://ray.yorksj.ac.uk/id/eprint/13203 |
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