ICAD Deficiency in Human Colon Cancer and Predisposition to Colon Tumorigenesis: Linkage to Apoptosis Resistance and Genomic Instability
Figure 5
ICAD-deficiency is associated with severe genomic instability in colon of DMH-treated mice.
Chromosomal DNA isolated from the different experimental groups was assessed for general chromosomal aberrations using an oligo microarray-based CGH with a chip containing 105,000 mouse genomic probes. DNA from colon tissue of age-matched naïve WT or ICAD−/− mice were used as reference controls. Test and reference DNA were labeled by random priming with Cy5-dUTP and Cy3-dUTP, respectively, using the Agilent Genomic DNA Labeling Kit Plus. The individually labeled test and reference samples were concentrated and then combined. Following probe denaturation and pre-annealing with mouse Cot-1 DNA, hybridization was performed as described in the methods. Data including Copy Number Variations were obtained by Agilent Feature Extraction software 9 and analyzed with Agilent Genomic Workbench software 5.0, using the statistical algorithms z score and ADM-2 according to sensitivity threshold respectively at 2.5 and 6.0 and a moving average window of 0.2 Mb. (A) Depiction of amplifications (pink) and deletions (green) throughout the 19 somatic chromosomes and the X and Y chromosomes. (B) Quantitative assessment of total aberrations (left panel); the right panel represents data attained when stringency was increased and a cutoff of a 3-fold change was applied; p<1×E−11.