Research Methods in Cancer Biology
Module code: MB7003
When a gene is found to be mutated in a cancer, we first need to understand how its protein product functions and then how the mutated protein alters the behaviour of cancer cells. For example, cancer cells show uncontrolled cell proliferation. It is therefore essential to learn techniques to study the function of cancer-causing or cancer-suppressing proteins and how changes to these proteins affects cells.
In the first practical you will use molecular techniques to identify a single nucleotide polymorphism (SNP) in a cancer-related gene -the total number of SNPs in the human genome is estimated to be more than 10 million! With recent advances in genomics, great effort is being directed towards the search for genetic variants (i.e., SNPs) that influence cancer susceptibility and outcome (Erichsen and Chanock, 2004).
In the second practical you will examine activation of the Mitogen Activated Protein Kinase (MAPK) pathway. This signal transduction pathway regulates a number of different cell functions such as growth, proliferation, differentiation, apoptosis and cell migration. Some components of the MAPK pathway, such as the growth factor receptors, Ras and Raf, act as oncoproteins in cancer cells and flood the cell with growth-stimulating signals. You will use SDS-PAGE, western blotting and GST pull-down assays to examine activation of the MAPK pathway.
As one of the key hallmarks of cancer cells is ‘unlimited replicative potential’ it is important to be able to use techniques to analyse the eukaryotic cell cycle. The third practical in this module will teach you some of the key methods that are used in cell cycle analysis including basic and advanced microscopy (light, confocal and electron microscopy), the use of epitope tags such as EGFP (Enhanced Green Fluorescent Protein) and antibodies to study the intracellular localisation of subcellular organelles and proteins, DNA replication using BrdU labelling and flow cytometry. Other key techniques you will learn in this practical include mammalian cell culture, transient transfection and immunoprecipitation.