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Title: Evaluation of in vitro models for detection of hepatocyte toxicity induced by anti-cancer drugs
Author: Reid, E.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2008
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The aim of the studies included in this thesis is to evaluate in vitro models in which anti-cancer drug-induced hepatoxicity may be detected, to decrease the likelihood of hepatotoxic anticancer agents energin clinical development. In vitro models that represent hepatocytes, the major cell type in the liver, were selected, and were all of human origin. Murine models were not considered as there are significant inter-species differences that may impact on drug metabolism and therefore on drug-induced liver toxicity. The selected models comprised fresh and cryopreserved hepatocytes (from 24 and 3 individuals respectively), an immortalised hepatocyte cell line (THLE-2) and hepatoma cell lines (Hep 3B2.1-7, Hep G2, Huh-7D12, PLC/PRF/5, and SK-HEP-1). These cells were characterised and then assessed in terms of their sensitivity to a known hepatoxic drug (mithramycin), drugs used in clinical practice in colorectal cancer (oxaliplatin, 5-FU) and novel agents in preclinical development (ruthenium compounds). A subset of in vitro models (fresh and cryopreserved hepatocytes, Hep3B2.1-7, HepG2, Huh-7D12 and PLC/PRF/5), which appeared to be the most promising based on gene expression and cytotoxicity data, were then selected to study the contribution of oncosis and/or apoptosis to cytotoxicity, and to see whether a hepatotoxic drug could be detected earlier during drug exposure. Assays to measure ATP levels and caspase-3/7 activation in drug-treated cells were optimised and used for mithramycin-, oxaliplatin- and 5-FU-treated cells. ATP levels and caspase-3/7 activation varied between cell lines and hepatocytes in response to all three drugs investigated, and was detectable immediately following drug treatment. These data suggest that fresh hepatocytes are a suitable in vitro model in which to accurately identify hepatotoxicity associated with anti-cancer drugs. They express liver-specific genes, and are more sensitive to the hepatotoxic drug mithramycin, than to oxaliplatin and 5-FU. A cytotoxicity assay using hepatoma cell lines (Hep3B2.1-7 and Huh-7D12) could also be used, despite their low expression of liver-related genes. These results should help select the most appropriate in vitro models to detect hepatotoxic anticancer agents in the future.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available