The main focus of my work is investigating genetic alterations underlying rectal carcinogenesis. Specifically, I would like to inhibit expression of genes that are highly expressed in carcinomas using gene silencing mechanisms. Conversely, we will be transfecting cells with expression constructs containing those genes that demonstrated reduced expression in the carcinomas relative to normal rectal mucosa.
 
Several genes were recently discovered to be differentially regulated between rectal carcinomas and normal rectal mucosa. I will utilize RNA interference to induce specific down-regulation of these genes. In order to evaluate the effectiveness of the siRNA strategy, I will assay the mRNA production by use of gene-specific probes. The siRNA protocols will be developed using well-characterized colorectal carcinoma cell lines: HCT 116, HT 29, and DLD 1.  
 
In addition, we plan to immortalize cell lines from the tumor and mucosa samples in which the initial differential gene expression was identified. The siRNA protocols developed with the established cell lines will then be applied to the new cell lines to determine their effectiveness in reversing the carcinogenic phenotype.