Gene expression signatures in anti-FGFR2IIIc monoclonal antibody-treated human colorectal cancer cells

To investigate the underlying mechanisms of the inhibitory effects of human anti-human FGFR2IIIc antibody on growth and migration of colorectal cancer cells, we used DNA microarray analysis to examine the cell signaling pathway alterations following the administration of anti-FGFR2IIIc antibody. Cells were plated at a density of 2.5 × 105 cells in a 60-mm dish, and grown overnight. Then 100 μg/mL of monoclonal anti-human FGFR2IIIc antibody was added in each dish. For control groups, an equal amount of anti-GFP antibody was added in another dish. After 48 hr, total RNA was isolated from cells. For use in DNA microarray analysis, 50 ng RNA from each group of cells was labeled using the Low Input Quick Amp Labeling kit. Labeled RNA was further purified using the Qiagen RNeasy Mini kit. Labeled cRNA was hybridized to the Agilent human 44k oligonucleotide microarray, and washed using Agilent Gene Expression washing buffer.