Ornithine decarboxylase antizyme inhibitor 2 (AZIN2) is a signature of secretory phenotype and independent predictor of adverse prognosis in colorectal cancer.

Tuomas Kaprio,Tiina Rasila,Jaana Hagström,Harri Mustonen,Petri Lankila,Leif C Andersson,Caj Haglund,Irina V Lebedeva

doi:10.1371/journal.pone.0211564

Abstract

Ornithine decarboxylase (ODC) is the rate-limiting enzyme of polyamine synthesis. The two ODC antizyme inhibitors (AZIN1) and (AZIN2) are regulators of the catalytic activity of ODC. While AZIN1 is a regulator of cell proliferation, AZIN2 is involved in intracellular vesicle transport and secretion. There are no previous reports on the impact of AZIN2 expression in human cancer. We applied immunohistochemistry with antibodies to human AZIN2 on tissue micro- arrays of colorectal cancers (CRC) from 840 patients with a median follow-up of 5.1 years (range 0–25.8). The 5-year disease-specific survival rate was 58.9% (95% Cl 55.0–62.8%). High AZIN2 expression was associated with mucinous histology (p = 0.002) and location in the right hemicolon (p = 0.021). We found no association with age, gender, stage, or histological tumor grade. High tumor expression of AZIN2 predicted an unfavorable prognosis (p<0.0001, log-rank test), compared to low AZIN2 expression. Cox multivariable analysis identified AZIN2 as an independent factor of an unfavorable prognosis in CRC. The strongest AZIN2 expression was seen in invasive tumor cells having morphological features of epithelial-mesenchymal transition (EMT). Induction of EMT in HT-29 CRC cells lead to upregulated expression of endogenous AZIN2. Given that AZIN2 is a regulator of vesicle transport and secretion, we overexpressed human AZIN2 cDNA in T84 CRC cells, and found strongly enhanced accumulation of CD63-positive exosomes in the culture medium. These findings indicate that AZIN2 expression is a signature of EMT-associated secretory phenotype that is linked to an adverse prognosis in CRC.

Highlights

Colorectal cancer (CRC), with over one million new cases every year, is one of the three most common cancers worldwide, and its incidence is rising
AZIN1 and AZIN2 are both positive activators of Ornithine decarboxylase (ODC)
AZIN1, which is physiologically involved in regulating the cell cycle and in stimulating cell proliferation, has been associated with cancer

Summary

Introduction

Colorectal cancer (CRC), with over one million new cases every year, is one of the three most common cancers worldwide, and its incidence is rising. High ODC activity is typically found in rapidly proliferating normal and malignant cells and in cancerous tissue. Immunohistochemical staining of sections of colon cancers with antibodies to human AZIN2 revealed its elevated expression in the invasive cells of the tumor fronts. This observation prompted us to investigate a tissue micro-array material consisting of 840 colorectal cancers in order to evaluate the prognostic role of AZIN2 expression, and its association with clinicopathological parameters. Based on our finding that AZIN2 is a regulator of vesicle transport and mast cell degranulation, we used a human colon cancer cell line to study the impact of AZIN2 expression on in vitro release of exosome-like material. Elevated plasma levels of circulating tumor-derived exosomes have been reported to correlate with poor prognosis and shorter survival time in CRC. [15]

Materials and methods

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Discussion