Research Methods in Cell Biology
Module code: MB7002
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 protein produced by the mutant gene 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. During a practical session 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, immunecomplex kinase assays 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 second practical in this module will teach students 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 and antibodies to study the intracellular localisation of subcellular organelles and proteins, BrdU labelling and flow cytometry. Other key techniques you will learn in this practical include mammalian cell culture, transient transfection and the identification of unknown proteins using mass spectrometry.