Volume 132, Issue 2, Pages 645-653 (February 2007) A Simple Method for the Routine Detection of Somatic Quantitative Genetic Alterations in Colorectal Cancer Audrey Killian, Frédéric Di Fiore, Florence Le Pessot, France Blanchard, Aude Lamy, Grégory Raux, Jean–Michel Flaman, Bernard Paillot, Pierre Michel, Jean–Christophe Sabourin, Jean–Jacques Tuech, Francis Michot, Jean–Pierre Kerckaert, Richard Sesboüé, Thierry Frebourg Gastroenterology Volume 132, Issue 2, Pages 645-653 (February 2007) DOI: 10.1053/j.gastro.2006.12.006 Copyright © 2007 AGA Institute Terms and Conditions
Figure 1 QMPSF-based bar code in primary colorectal cancers. The QMPSF profile generated from malignant tissue (red) was superimposed on that obtained from distant nonmalignant tissue (blue) by adjusting to the same height the 2 control amplicons indicated in green. Each peak corresponds to a target gene. The heights of red and blue peaks corresponding to a specific target gene were then compared. The y-axis displays fluorescence intensity, and the x-axis indicates the amplicon size in base pairs. (A) Tumor with normal gene dosage. (B) Tumor showing gene dosage alteration of several genes (loss of copy for DCC, TP53, and BLK; gain of copy for EGFR, MYC, and AURKA). Arrows indicate the top of blue or red peaks when gene dosage is altered. Gastroenterology 2007 132, 645-653DOI: (10.1053/j.gastro.2006.12.006) Copyright © 2007 AGA Institute Terms and Conditions
Figure 2 QMPSF-based kinogram in primary colorectal cancers. The QMPSF profile generated from malignant tissue (red) was superimposed on that obtained from distant nonmalignant tissue (blue) by adjusting to the same height the 2 control amplicons indicated in green. Each peak corresponds to a target gene. The heights of red and blue peaks corresponding to a specific target gene were then compared. The y-axis displays fluorescence intensity, and the x-axis indicates the amplicon size in base pairs. Example of a tumor showing an increase of gene copy number affecting 5 kinase genes: EGFR, MET, FGFR1, PTK2, and AURKA. Arrows indicate the top of blue peak. Gastroenterology 2007 132, 645-653DOI: (10.1053/j.gastro.2006.12.006) Copyright © 2007 AGA Institute Terms and Conditions
Figure 3 Comparison of gene dosage analysis performed by quantitative measurement on 23 targets in 3 CRC samples using QMPSF (x-axis) and aCGH (y-axis). (A) Sample (GS03.019) with normal gene copy numbers. (B and C) Samples with altered gene copy numbers (GS04.252, GS04.254). The regression lines are only indicative. Gastroenterology 2007 132, 645-653DOI: (10.1053/j.gastro.2006.12.006) Copyright © 2007 AGA Institute Terms and Conditions
Figure 4 Comparison of the results obtained by QMPSF and aCGH on a CRC sample (GS04.252). (A) Detection of an RAF1 copy number loss. (B) Detection of an AURKA gain of copy number. Arrows on the QMPSF profile (left) indicate the top of altered peaks. Each dot on the array CGH profile (right) corresponds to 1 probe; the color indicates ratios corresponding to the following: normal level (0.84–1.19) in black, decrease (<0.84) in green, or increase (>1.19) in red. The boxes outline the probes corresponding to the genes under study. Gastroenterology 2007 132, 645-653DOI: (10.1053/j.gastro.2006.12.006) Copyright © 2007 AGA Institute Terms and Conditions