Establishment of new predictive markers for distant recurrence of colorectal cancer using lectin microarray analysis

Abstract

We evaluated the clinical benefits of novel predictive markers for distant recurrence with colorectal cancer using lectin microarray analysis of cell surface glycan modifications. Glycoproteins were extracted from formalin-fixed, paraffin-embedded tumor specimens and normal epithelium from 53 consecutive curatively resected stage I-III colorectal cancer cases and then subjected to lectin microarray to obtain lectin-glycan interaction (LGI) values. In addition, clinicopathological factors associated with distant recurrence were identified. LGI values that were associated with distant recurrence were validated with an additional 55 curatively resected stage II colorectal cancer cases. LGI values for Agaricus bisporus (ABA) lectin, prominent in cancer tissues, were statistically associated with distant recurrence. ABA lectin staining exhibited strikingly intense signals in the cytoplasm and apical surfaces of cancer cells, while weak staining was observed in the supranuclear regions of normal epithelium. This ABA tumor/normal LGI ratio may be a new predictive biomarker for distant recurrence of curatively resected colorectal cancer.

Keywords: Colorectal cancer; glycoprotein; lectin; prediction; recurrence.

Figure 1

β-actin staining. β-actin in the colorectal cancer tissues (A) or normal colorectal tissues (C) were visualized with anti-human β-actin mAb and biotin–avidin–peroxidase method. Hematoxylin staining of the colorectal cancer tissues (B) or normal colorectal tissues (D) as a control were also performed (×100).

Figure 2

Recurrence-free survival analysis of stage I–III colorectal cancer patients (n = 53) with Agaricus bisporus (ABA) and Hippeastrum hybrid (HHL) status. (A) Five-year recurrence-free survival were 82.0% and 24.7% for the patients with ABA <2.18, and ABA ≥2.18 (P = 0.001). (B) Five-year recurrence-free survival were 88.8% and 40.9% for the patients with HHL <4.00, and HHL ≥4.00 (P = 0.001).

Figure 3

Recurrence-free survival analysis of stage II colorectal cancer patients (n = 55) with Agaricus bisporus (ABA) status in the validation setting. Recurrence-free survival according to ABA status (P = 0.003).

Figure 4

Agaricus bisporus (ABA) staining for colorectal cancer tissues. (A) Strong ABA staining was detected within the cytoplasm of cancer cells and the apical surface (×100, ×400). (B) ABA staining was detected in the supranuclear region of the glandular epithelium of normal colorectal tissues (×100, ×400).