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The general theme of the Thomas Ried Laboratory at the National Cancer Institute is to understand the role of genomic instability and associated gene expression and protein changes during cancer development. Toward this goal, we study models of human tumorigenesis and relevant mouse models with a variety of genomic technologies, including comparative genomic hybridization, spectral karyotyping (SKY), gene expression profiling, real-time PCR, and proteomic methodologies. Our translational research is focused on exploring aneuploidy as a molecular marker for cancer diagnostics in routine cytological preparations and to identify gene expression and protein signatures that assist in disease prognosis and rational cancer therapy.
www.riedlab.nci.nih.gov
The Technique
In the Ried Laboratory, we are interested in understanding the genomic, transcriptomic, and proteomic changes that accompany the development of cancer. The coordinated, simultaneous extraction of DNA, RNA, and proteins from a single sample is crucial for accurate correlations between genomic aberrations and their consequences on the transcriptome and proteome. Such correlations obviously benefit from using identical tissue samples for simultaneous analysis of DNA, RNA, and proteins. Toward this end, we developed a method that allows complete recovery of proteins from a sample already harvested for RNA and DNA using TRIZOL®. With this method, we can use a single sample to obtain information about changes in the genomic copy number, gene, and protein expression levels. We have even been able to demonstrate that the method works on small biopsy samples and samples that have been stored at −20°C for several years. This technique will allow us to begin a systematic exploration of genomic alterations, their consequences on gene expression levels, and ultimately, on the proteome.
Isolation and solubilization of proteins after TRIZOL® extraction of RNA and DNA from patient material following prolonged storage, p. 467.
