Volume 127, Issue 2, Pages (August 2004)

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Volume 127, Issue 2, Pages 457-469 (August 2004) Expression profiling identifies chemokine (C-C motif) ligand 18 as an independent prognostic indicator in gastric cancer Suet Yi Leung, Siu Tsan Yuen, Kent-Man Chu, Jonathan A. Mathy, Rui Li, Annie S.Y. Chan, Simon Law, John Wong, Xin Chen, Samuel So Gastroenterology Volume 127, Issue 2, Pages 457-469 (August 2004) DOI: 10.1053/j.gastro.2004.05.031

Figure 1 Expression of a leukocyte gene cluster in human gastric tissues. (A) Hierarchical clustering of 6688 cDNA clones (corresponding to 5200 unique genes) in 126 gastric tissue samples, including gastric tumors (n = 90), metastasis to lymph nodes (n = 14), and nontumor gastric mucosa (n = 22). Data are presented in table format, where each row represents a gene and each column represents a sample. The relative abundance of each transcript centered across all the samples is depicted according to the color scale shown in the lower right corner. Gray indicates missing or excluded data. Specific gene clusters identified in the gastric tissue expression profiles are annotated to the right. See Chen et al.5 for a detailed analysis of gene-expression patterns in human gastric cancers. The unabridged data are available through our Web site at http://genome-www.stanford.edu/Gastric_Cancer2. (B) Expanded view of a portion of data shown in (A), represented by the purple bar to the left, reflecting genes belonging to the leukocyte gene cluster. Because of limited space, only a few selected genes are depicted. (A) is the same as previously published4,5 and is reproduced with permission from the American Society for Cell Biology. ECM, extracellular matrix. Gastroenterology 2004 127, 457-469DOI: (10.1053/j.gastro.2004.05.031)

Figure 2 Correlation between the CCL18 expression levels measured by cDNA microarray and real-time quantitative RT-PCR. Gastroenterology 2004 127, 457-469DOI: (10.1053/j.gastro.2004.05.031)

Figure 3 (A and B) Kaplan-Meier plot of overall and disease-free survival in gastric cancer patients expressing high or low levels of CCL18, as assayed by cDNA microarray. Results of the log-rank test are shown at the right upper corner. (C and D) Kaplan-Meier plot of overall and disease-free survival in an independent set of gastric cancer patients expressing high or low levels of CCL18, as assayed by quantitative RT-PCR. Gastroenterology 2004 127, 457-469DOI: (10.1053/j.gastro.2004.05.031)

Figure 4 Immunohistochemical (IHC) detection of CCL18 expression in human gastric cancer tissues. (A) A gastric cancer (HKG73T) showing abundant CCL18+ cells (IHC grade 3), especially at the tumor invasion front. (Right) Expanded view showing macrophage morphology of the CCL-18+ cells. (B) Consecutive section of (A) showing abundant CD68+ tumor-associated macrophages (IHC grade 3). (C) Double staining with anti-CD68 antibody (red) and anti-CCL18 antibody (brown), showing CCL18 expression in a subset of macrophages within human gastric cancer tissues (HKG73T). (Right) Expanded view showing CCL18 expression in a subset of macrophages. Note that the lower 2 cells have both brown and red staining, whereas the upper 2 cells show only red staining (anti-CD68). (D) Nonneoplastic gastric mucosa with active chronic gastritis and Helicobacter pylori infection. Only a single CCL18+ cell (arrow) can be seen in this field. Other areas with light brown staining are due to nonspecific uptake by red blood cells. (E) A gastric cancer (HKG78T) with no CCL18-expressing cells (IHC grade 1). (F) Consecutive section of (E) showing a moderate number of CD68+ macrophages in the tumor (IHC grade 2). Gastroenterology 2004 127, 457-469DOI: (10.1053/j.gastro.2004.05.031)