The use of novel diagnostic cell cycle markers and targeted treatment of pancreaticobiliary cancers

Adenocarcinoma of the pancreas is one of the top ten leading causes of cancer deaths. In the UK approximately 8000 people are diagnosed with the disease each year. Surgical resection is the only chance of cure and is only possible in a minority of patients. Even after resection, median survival is only 10–20 months and no more than 5–20% of resected patients survive five years. Earlier diagnostic assays and novel treatments are urgently required to improve outcomes. Chapter 1 summarises clinical aspects of pancreatic cancer and current diagnostic and prognostic tests. Chapter 2 gives an introduction to the cell cycle, initiation of DNA replication, cell cycle checkpoints and how exploitation of these mechanisms can result in selective death of cancer cells. Chapter 3 describes an analysis of four cell cycle proteins - Mcm2, geminin, phosphohistone H3 and Cdc7 - using immunohistochemistry of archived FFPE tissue from patients with pancreatic cancer. Linked clinicopathological variables were used to attempt to predict outcome and to validate Cdc7 as a potential target in the treatment of pancreatic cancer. Chapter 4 describes cell culture techniques that were used to grow pancreatic adenocarcinoma and IMR-90 fibroblast cell lines in the laboratory. Cells were treated with siRNA against CDC7 and a small molecule inhibitor of Cdc7/Cdk9. A variety of techniques including FACS analysis, western blotting, BrdU labelling and apoptosis assays were used to assess the effects of these treatments. Chapter 5 describes the methods and early results obtained from using a novel Mcm5 assay for the determination of malignancy in biliary brushings. Chapter 6 discusses the main conclusions drawn from the work as well as potential future studies