New markers of pancreatic cancer identified through differential gene expression analyses: claudin 18 and annexin A8

Abstract

Background: New markers to distinguish benign reactive glands from infiltrating ductal adenocarcinoma of the pancreas are needed.

Design: The gene expression patterns of 24 surgically resected primary infiltrating ductal adenocarcinomas of the pancreas were compared with 18 non-neoplastic samples using the Affymetrix U133 Plus 2.0 Arrays and the Gene Logic GeneExpress Software System. Gene fragments from 4 genes (annexin A8, claudin 18, CXCL5, and S100 A2) were selected from the fragments found to be highly expressed in infiltrating adenocarcinomas when compared with normal tissues. The protein expression of these genes was examined using immunohistochemical labeling of tissue microarrays.

Results: Claudin 18 labeled infiltrating carcinomas in a membranous pattern. When compared with normal and reactive ducts, claudin 18 was overexpressed, at least focally, in 159 of 166 evaluable carcinomas (96%). Strong and diffuse claudin 18 overexpression was most often seen in well-differentiated carcinomas (P=0.02). Claudin 18 was overexpressed in 51 of 52 cases (98%) of pancreatic intraepithelial neoplasia. Annexin A8 was at least focally overexpressed in 149 of 154 evaluable infiltrating carcinomas (97%). S100 A2 was at least focally overexpressed in 118 of 154 evaluable infiltrating carcinomas (77%). Non-neoplastic glands also frequently expressed S100 A2 diminishing its potential diagnostic utility. Immunolabeling with antibodies directed against CXCL5 did not reveal any significant differences in protein expression between infiltrating adenocarcinomas and normal pancreatic ducts.

Conclusions: Claudin 18 and annexin A8 are frequently highly overexpressed in infiltrating ductal adenocarcinomas when compared with normal reactive ducts, suggesting a role for these molecules in pancreatic ductal adenocarcinomas. Furthermore, these may serve as diagnostic markers, as screening tests and as therapeutic targets.

FIGURE 1

Immunohistochemical labeling for claudin 18 demonstrates strong, membranous labeling of the infiltrating carcinoma and absence of labeling of the non-neoplastic glands and stroma (A–D).

FIGURE 2

Kaplan-Meier survival curves for patients with pancreatic ductal adenocarcinoma after pancreaticoduodenectomy based on claudin 18 expression. Patients whose carcinomas strongly and diffusely labeled with the antibody to claudin 18 had a significantly better survival than did patients whose carcinomas weakly labeled or did not label, even when controlling for tumor grade (log-rank P = 0.013).

FIGURE 3

Immunohistochemical labeling for claudin 18 labels (A) PanIN-2 and (B) PanIN-3 precursor lesions. The pattern of labeling is membranous and is similar to the labeling seen in PanIN-1 lesions (not shown).

FIGURE 4

Immunohistochemical labeling for annexin A8 demonstrates strong and diffuse labeling of the neoplastic glands (A–D). The adjacent lymphocytes, reactive glands, and stroma do not label.