Hypoxia-Related Marker GLUT-1, CAIX, Proliferative Index and Microvessel Density in Canine Oral Malignant Neoplasia

Valeria Meier; Franco Guscetti; Malgorzata Roos; Stefanie Ohlerth; Martin Pruschy; Carla Rohrer Bley

doi:10.1371/journal.pone.0149993

Hypoxia-Related Marker GLUT-1, CAIX, Proliferative Index and Microvessel Density in Canine Oral Malignant Neoplasia

PLoS One. 2016 Feb 23;11(2):e0149993. doi: 10.1371/journal.pone.0149993. eCollection 2016.

Authors

Valeria Meier¹, Franco Guscetti², Malgorzata Roos³, Stefanie Ohlerth⁴, Martin Pruschy⁵, Carla Rohrer Bley¹

Affiliations

¹ Division of Radiation Oncology, Vetsuisse Faculty University of Zurich, Zurich, Switzerland.
² Institute of Veterinary Pathology, University of Zurich, Zurich, Switzerland.
³ Department of Biostatistics, Epidemiology Biostatistics and Prevention Institute, Faculty of Medicine, University of Zurich, Zurich, Switzerland.
⁴ Clinic of Diagnostic Imaging, Vetsuisse Faculty University of Zurich, Zurich, Switzerland.
⁵ Laboratory for Molecular Radiobiology, Radiation Oncology, University Hospital Zurich, Zurich, Switzerland.

Abstract

For various types of tumor therapy, it is suggested that co-targeting of tumor microenvironment, mainly tumor vasculature, mediates tumor response mechanisms. Immunohistochemistry for glucose transporter-1 (GLUT-1), carbonic anhydrase-IX (CAIX), Ki-67, and von Willebrand factor VIII for microvessel density (MVD) were performed on formalin-fixed paraffin-embedded samples of canine oral malignant neoplasms. Polarographic oxygen measurements (median pO2) and perfusion data via contrast-enhanced power Doppler ultrasound (median vascularity, median blood volume) provided additional information. Ninety-two samples were analyzed: sarcomas (n = 32), carcinomas (n = 30), and malignant melanomas (n = 30). Polarographic oxygen and perfusion data was available in 22.8% (sarcomas n = 9, carcinomas n = 7, melanomas n = 5), and 27.1% (sarcomas n = 10, carcinomas n = 8, melanomas n = 7) of cases, respectively. GLUT-1 expression was detected in 46.7% of all samples, and was generally weak. CAIX expression was found in 34.8% of all samples. Median Ki-67 score and MVD count was 19% and 17, respectively. The evaluation of the GLUT-1 score and continuous data showed significantly lower GLUT-1 levels in sarcomas (mean 5.1%, SD 6.2) versus carcinomas and melanomas (mean 16.5%/ 19.0%, SD 17.3/ 20.9, p = 0.001). The expression of CAIX correlated mildly positively with GLUT-1 (p = 0.018, rho = 0.250) as well as with Ki-67 (p = 0.014, rho = 0.295). MVD showed a significantly lower level in melanomas (mean 12.6, SD 7.7) versus sarcomas and carcinomas (mean 21.8/ 26.9, SD 13.0/20.4, p = 0.001). Median vascularity and blood volume were significantly lower in sarcomas (mean 10.4%, SD 11.0, and mean 6.3%, SD 6.5, respectively) versus carcinomas (mean 39.2%, SD 16.4 and mean 33.0%, SD 25.6, respectively) and melanomas (mean 36.0%, SD 18.3, and 31.5%, SD 24.5). Between the 3 histological groups, there was neither a significant difference in the GLUT-1 and CAIX score and continuous data, nor the Ki67 score, or polarographic oxygen measurements. GLUT-1 continuous data and Ki-67 (p<0.001, rho = 0.403), as well as Ki-67 and MVD (p = 0.029, rho = 0.228) correlated positively and a mild correlation was found between vascularity and GLUT-1 (p = 0.043, rho = 0.408). GLUT-1, CAIX, proliferative index and MVD levels were established as microenvironmental descriptors with the purpose of creating a baseline in order to follow changes seen in the tumor microenvironment after hypofractionated radiation with high doses.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Carbonic Anhydrases / biosynthesis*
Cell Hypoxia
Cell Proliferation*
Dogs
Gene Expression Regulation, Neoplastic*
Glucose Transporter Type 1 / biosynthesis*
Microcirculation*
Mouth Neoplasms* / blood supply
Mouth Neoplasms* / metabolism
Mouth Neoplasms* / pathology
Neoplasm Proteins / biosynthesis*

Substances

Glucose Transporter Type 1
Neoplasm Proteins
Carbonic Anhydrases

Grants and funding

This study was funded by the Swiss National Science Foundation (SNSF; recipient: CRB, grant number: 320030_132441, URL: http://www.snf.ch). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.