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Volume 122, Issue 2, Pages 376-386 (February 2002) Aminopeptidase N is involved in cell motility and angiogenesis: Its clinical significance in human colon cancer Hiroki Hashida, Arimichi Takabayashi, Michiyuki Kanai, Masashi Adachi, Keiichi Kondo, Nobuoki Kohno, Yoshio Yamaoka, Masayuki Miyake Gastroenterology Volume 122, Issue 2, Pages 376-386 (February 2002) DOI: 10.1053/gast.2002.31095 Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 1 Dose-dependent effect of mAb MH8-11 on cell penetration. Cell penetration was determined with HT1080-penetrating cells into the lower chamber of a Transwell. A constant inhibitory effect was produced by an mAb MH8-11 concentration of 5 μg/mL. Gastroenterology 2002 122, 376-386DOI: (10.1053/gast.2002.31095) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 2 Effect of mAb MH8-11 on the motility of HT1080 cells as determined by a phagokinetic assay using the gold colloid method. (A) HT1080 cell motility in medium containing mouse IgG (5 μg/mL). (B) HT1080 cell motility in medium containing MH8-11 (5 μg/mL). Gastroenterology 2002 122, 376-386DOI: (10.1053/gast.2002.31095) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 3 Effect of mAb MH8-11 on EC migration. The number of migrated HUVECs through the filter coated by reconstituted basement membrane was counted. HUVEC migration was inhibited more significantly by functional mAb MH8-11 (5 μg/mL; P < 0.001). Gastroenterology 2002 122, 376-386DOI: (10.1053/gast.2002.31095) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 4 Effect of mAb MH8-11 on HUVEC tube formation (original magnification, 200×). (A) HUVEC tube formation in medium containing mouse IgG (5 μg/mL). (B) HUVEC tube formation in medium containing mAb MH8-11 (5 μg/mL). Gastroenterology 2002 122, 376-386DOI: (10.1053/gast.2002.31095) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 5 Effect of mAb MH8-11 on HUVEC tube (A) width and (B) length. HUVECs were plated onto Matrigel in the presence of mAb MH8-11 (5 μg/mL). Each experiment showed a similar and significant decrease in the presence of mAb MH8-11 (P < 0.001 and < 0.001, respectively). Gastroenterology 2002 122, 376-386DOI: (10.1053/gast.2002.31095) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 6 Western blotting analysis of APN/CD13 expression. Arrowhead indicates 165-kD molecular weight. Gastroenterology 2002 122, 376-386DOI: (10.1053/gast.2002.31095) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 7 Immunohistochemical staining of human colon cancer tissues using the avidin-biotin-peroxidase complex procedure (original magnification 150×). (A) APN/CD13-positive staining of colon cancer. (B) APN/CD13-negative staining of colon cancer. Gastroenterology 2002 122, 376-386DOI: (10.1053/gast.2002.31095) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 8 (A) Overall survival of 114 colon cancer patients according to their tumor APN/CD13 status. (B) Overall survival of 42 node-positive colon cancer patients according to their tumor APN/CD13 status. Gastroenterology 2002 122, 376-386DOI: (10.1053/gast.2002.31095) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 9 (A) Disease-free survival of 114 colon cancer patients according to their tumor APN/CD13 status. (B) Disease-free survival of 42 node-positive colon cancer patients according to their tumor APN/CD13 status. Gastroenterology 2002 122, 376-386DOI: (10.1053/gast.2002.31095) Copyright © 2002 American Gastroenterological Association Terms and Conditions