Serum Level of Tumor-Overexpressed AGR2 Is Significantly Associated with Unfavorable Prognosis of Canine Malignant Mammary Tumors.

Stephen Hsien-Chi Yuan,Shih-Chieh Chang,Hao-Ping Liu,Yenlin Huang

doi:10.3390/ani11102923

Abstract

Simple SummaryCanine malignant mammary tumor (MMT) is a prevalent malignancy in intact female dogs. A current lack of easily accessible tumor biomarkers hinders a timely assessment of the disease outcome. This study reveals that anterior gradient protein 2 (AGR2) is overexpressed in canine MMT tissues, and elevated levels of extracellular AGR2 in sera of MMT dogs are significantly associated with progression and remote metastasis of MMT and an unfavorable overall survival of the patients. Hence, serum eAGR2 level is significantly associated with an adverse outcome of MMT dogs and holds a predictive potential in MMT prognosis.Canine malignant mammary tumors (MMTs) are prevalent malignancy in intact female dogs with a high incidence of metastasis and recurrence. A current lack of easily accessible tumor biomarkers hinders a timely assessment of the disease outcome. We previously identified anterior gradient protein 2 (AGR2) with higher protein abundance in canine MMT tissues compared with normal counterparts. AGR2 is an endoplasmic reticulum-resident protein disulfide isomerase involved in the regulation of protein processing and also exists extracellularly via secretion to exert pro-oncogenic functions. In the present study, we validated overexpression of AGR2 in canine MMT tissues from 45 dogs using immunohistochemistry and immunoblotting, and assessed serum AGR2 levels in 81 dogs with MMTs and 21 benign cases using a competitive enzyme-linked immunosorbent assay (ELISA). Our data revealed that serum eAGR2 levels are significantly correlated with MMT progression (p = 0.0007) and remote tumor metastasis (p = 0.002). Moreover, elevated levels of serum eAGR2 are associated with an unfavorable overall survival of MMT dogs in later stage (p = 0.0158). Area under the time-dependent ROC curve (AUC) of serum eAGR2 level as a prognostic indicator was 0.839. Collectively, this study uncovered that serum eAGR2 level is significantly associated with an adverse outcome of MMT dogs and holds a predictive potential in MMT prognosis.

Highlights

Canine mammary tumors (MTs) are one of the most prevalent neoplasms in intact female dogs, approximately 50% of which are malignant [1]
2A,C), in consistence with the Collectively, anterior gradient protein 2 (AGR2) was in overall mammary tumors (MMTs) tissues and even in Benign Mammary Tumor (BMT) tissues compared with the normal counterparts overexpressed in overall
These data indicated a significant of AGR2 expression level with counterparts canine MMT.(Figure 2B)

Summary

Introduction

Canine mammary tumors (MTs) are one of the most prevalent neoplasms in intact female dogs, approximately 50% of which are malignant [1]. Most of the malignant MTs (MMTs) are of epithelial origin and classified into several subtypes. Prospective studies show that the MMT-associated death after 2 years of follow-up is 20–45% [2]. To identify new canine MMT-associated proteins that may contribute to the pathogenesis of this disease, we exploited quantitative proteomics analysis of canine MMT tissues and identified a list of proteins with higher 4.0/). Abundance in canine MMT tissues compared with normal counterparts, including anterior gradient 2 (AGR2) [3]. Proteins destined to the secretory pathway or to the cell surface are synthesized in the ER, where they undergo post-translational modification and proper folding executed by PDIs. For instance, production of the intestinal mucin MUC2 requires AGR2 via its thioredoxin-like domain to form intermolecular disulfide bonds with MUC2 [6]

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