Tyronsine Kinase Gene Expression Profiling in Prostate Cancer

摘要

The expression of protein kinases genes if often altered in tumors. The aberrant expression of several of these genes typically parallels the progression toward a more malignant phenotype. We developed a cDNA micro-array- based screening system to mearuser the level of expression of tyronsine kinase (tk) genes. The hardware for preparation of cDNA micro-arrays and basic protocols for hybridization were developed in year 1. In the second year, we finished cDNA synthesis from prostate cancer cell lines and 6 frozen tissue specimens. We continued our DNA sequencing effort and added additional targets to our micro-arrays. Using prostate cancer cell lines, the system delivered reproducible results about tk gene expression during cell transformation and progression toward a more malignant phenotype. Comparing the absolute expression levels form cDNA micro-arrays with data from Northern blot analyses suggested that our initial approach using mixed-based oligonucleotide primers led to lowered representation of highly abundant transcript. This problem has been addressed with a new primer design to be better suited to investigate the tk gene expression in small samples. We then applied our protocols to the gene expression analysis of micro-dissected tissues, and found reliable detection of abl gene overexpression with an RNA equivalent of 3 cells.