Expression Profiling of Tyrosine Kinase Genes.

摘要

Aberrant expression of receptor or cytosolic tyrosine kinase genes and, in particular, their hyper-expression are common phenomena in breast cancer, which are believed to alter cell growth and response to external signals such as growth factors, hormones etc. Knowledge about the relative levels of expression of many tyrosine kinase genes, all at the same time, might contribute significantly to a better understanding of the processes of tumor development and progression. We are developing a rapid assay that will use innovative cDNA micro-arrays carrying small amounts of individual tyrosine kinase gene-specific targets to simultaneously determine the expression levels of up to 100 tyrosine kinase genes using a small number of cells. Three years of research and development will lead to discovery of a set of gene-specific markers associated with breast cancer progression and a simple device capable of performing inexpensive expression profiling of these markers. The research and development efforts in the first year of this 3-year project focused on the design and testing of robotic instruments to prepare DNA micro-arrays and the preparation of prototype arrays carrying sets of more than 50 gene-specific tyrosine kinase fragments. The second year effort (i.e., the reporting period 1 Aug 2000-31 Jul 2001) was directed towards the optimization of cDNA preparation, labeling, hybridization and detection protocols as well as the molecular cloning and sequencing of breast cancer-specific tyrosine kinase gene transcripts.