Analysis of Legumain protein expression by immunohistochemistry in normal gastric mucosa, gastric cancer and lymph node metastasis.

<p>(a) (b) Legumain expression exhibited negative or diffuse weakly positive staining in the cytoplasm in distant normal mucosa. (c) Legumain exhibited strong vesicular positivity in the cytoplasm in primary gastric cancer compared with those of adjacent normal mucosa. Vesicles were scattered in the cytoplasm. (d). Legumain also overexpressed in metastasis lymph nodes. (magnification ×100).</p

Legumain expression at the mRNA and protein levels.

<p>(a) Western blotting analysis of Legumain in normal gastric cell lines (GES-1) and gastric cancer cell lines (MKN28, AGS, SGC-7901, MGC-803, BGC-823). BGC-823, MGC-803 cell (poorly differentiated) and AGS, SGC-7901 (moderately differentiated) showed lower expression of Legumain in comparison with GES-1 (normal gastric cell lines) and MKN28 (well differentiated), (b) β-actin is the loading control. Legumain expression was confirmed in all the cell lines by realtime PCR.</p

Expression of Legumain in normal colonic mucosa, cancerous tissue and lymph node metastasis.

<p>P values are based on Fisher’s exact test. Multiple comparisons between groups were determined with p = 0.01 adjustment, due to multiple hypotheses.</p>†<p>Significant difference in the expression of Legumain between noncancerous mucosa and the gastric cancerous tissue sample.</p>‡<p>Significant difference in the expression of Legumain between noncancerous mucosa and the Lymphnode tissue sample.</p

Association between Legumain expression and clinicopathologic factors in gastric cancers.

<p>P values are based on X² test,</p>*<p>Significant difference were determined with p<0.05.</p

Univariate and multivariate analysis of overall survival after surgery.

<p>HR: hazard radio, CI: confidence interval,</p>*<p>Significant difference.</p

Correlation of Legumain expression with survival curves of patients with gastric cancer.

<p>Patients with Legumain-positive expression showed significantly better disease-free survival (a) and overall survival (b) than those with Legumain-negative expression (P<0.001, log-rank test).</p