Stool screening for colorectal cancer: Molecular approaches

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Stool screening for colorectal cancer: Molecular approaches Neal K. Osborn, David A. Ahlquist Gastroenterology Volume 128, Issue 1, Pages 192-206 (January 2005) DOI: 10.1053/j.gastro.2004.10.041 Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 1 Variable occult hemoglobin levels in stools collected over 2 weeks from 12 patients with asymptomatic advanced colorectal cancer as determined by the quantitative HemoQuant and the qualitative Hemoccult test. The shaded zone represents the conventional normal range. Reprinted from Ahlquist et al10 with permission. Gastroenterology 2005 128, 192-206DOI: (10.1053/j.gastro.2004.10.041) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 2 MUCI immunostaining of resected colorectal adenocarcinoma. Note that immunoreactivity (reddish-brown) is confined to membrane surface and secreted mucus; adjacent normal epithelium remains nonreactive. Reprinted from Limburg et al50 with permission. Gastroenterology 2005 128, 192-206DOI: (10.1053/j.gastro.2004.10.041) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 3 MUCI stool concentrations as determined by immunoassays with 2 different anti-MUC1 antibodies: ELSA-CA 15-3 and Truquant BR immunoassays. Reprinted from Limburg et al50 with permission. Gastroenterology 2005 128, 192-206DOI: (10.1053/j.gastro.2004.10.041) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 4 Cytokeratin immunostaining of normal colonic epithelium. Note cytokeratin immunostaining within macrophages in the lamina propria suggesting phagocytosis of epithelial cells. Reprinted from Ahlquist et al62 with permission. Gastroenterology 2005 128, 192-206DOI: (10.1053/j.gastro.2004.10.041) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 5 Cytokeratin immunostains of surface epithelium and overlying mucus from (A) colorectal cancer and (B) normal colon. Note luxuriant population of viable-appearing colonocytes exfoliated into mucus over cancer but the hypocellular appearance of mucus over normal epithelium. Reprinted from Ahlquist et al62 with permission. Gastroenterology 2005 128, 192-206DOI: (10.1053/j.gastro.2004.10.041) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 6 Schematic of cellular fluxes at the surface of (A) colorectal cancer and (B) normal colorectal epithelium. Gastroenterology 2005 128, 192-206DOI: (10.1053/j.gastro.2004.10.041) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 7 Colonocytes isolated from stools of patients with colorectal cancer using (A) density gradient centrifugation and (B) immunobead capture techniques. In both examples, immunostaining with an antibody (PCA 31.1) recognizing a tumor-associated antigen was used (immunoreactivity reflected by dark grey color). Note cluster of malignant colonocytes adjacent to a benign squamous epithelial cell in panel (A). Gastroenterology 2005 128, 192-206DOI: (10.1053/j.gastro.2004.10.041) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 8 Loss of fecal colonocytes during stool storage. Data represent recovery of HT-29-cultured colonocytes spiked into normal human stools. Reprinted from Davies et al78 with permission. Gastroenterology 2005 128, 192-206DOI: (10.1053/j.gastro.2004.10.041) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 9 Point mutational results for APC codon 1378 on 6 patient stool DNA samples (5–10 and 15–20) analyzed in duplicate (lanes a + b). Wild-type reactions (upper gels) confirm capture of human DNA. Mutant reactions (lower gels) are positive in a patient with colorectal cancer (gel 18) and in positive controls (gels 12–14). Reprinted from Ahlquist et al61 with permission. Gastroenterology 2005 128, 192-206DOI: (10.1053/j.gastro.2004.10.041) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 10 Polyacrylamide gel electrophoresis analysis for deletions within Bat-26. Results representing 10 unique stool DNA samples (lanes 1–10) analyzed for deletions within Bat-26. The upper (U) region of the gel contains reaction products representing the wild-type, full-length product. Presence of reaction products within the lower (L) region of the gels indicates deletions within the Bat-26 poly A tract sequence. Samples in lanes 2 and 9 were obtained from patients with proximal colon cancer and gels 12–14 represent positive controls. Reprinted from Ahlquist et al61 with permission. Gastroenterology 2005 128, 192-206DOI: (10.1053/j.gastro.2004.10.041) Copyright © 2005 American Gastroenterological Association Terms and Conditions