Functional characterization of pancreatic cancer candidate genes

Coordinator:    PD Dr. Malte Buchholz
Institution: Philipps-Universität Marburg, Klinik für Innere Medizin SP Gastroenterologie
In previous high-throughput gene expression analyses of pancreatic cancer tissues and cell lines, we have identified a total of ~2000 pancreatic cancer candidate genes, which have subsequently been further characterized using serial hybridizations with candidate gene arrays. Following bioinformatic evaluation of the expression profiles and functional annotation of the gene products, 80 candidate genes wer selected for further functional analyses in a highly parallel fashion using the reverse transfection array technology. The principle of this technology is based on the ‘spotting’ of fluorescently tagged expression constructs on microscope glass slides, which are subsequently seeded with cell cultures of cancer and control cells, so that the target genes are expressed in the cells at the specific locations on the arrays. With these arrays, a host of functional assays, using among others specific antibody stainings, were performed in order to detect changes for example in growth characteristics, migratory potential, viability and differentiation of the cells in response to the altered gene expression. In total, 14 genes were selected for further detailed investigations. A first manuscript describing the role of the candidate gene Galectin-3 in pancreatic cancer has already been published. Furthermore, far-reaching and previously unknown oncogenic functions were documented for candidate genes Cofilin-1 and FASTK. Moreover, in vivo studies with implanted tumor cells revealed significantly reduced tumor growth after inactivation of either of these genes in pancreatic cancer cells. Further analyses to identify mechanisms and pathways which mediate the growth inhibitory effects of the inactivation of these genes as well as further work on the other candidate genes will continue after the end of this project.
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