Multiplex Gene Expression Profiling of 16 Target Genes in Neoplastic and Non-Neoplastic Canine Mammary Tissues Using Branched-DNA Assay

Florenza Lüder Ripoli; Susanne Conradine Hammer; Annika Mohr; Saskia Willenbrock; Marion Hewicker-Trautwein; Bertram Brenig; Hugo Murua Escobar; Ingo Nolte

doi:10.3390/ijms17091589

Multiplex Gene Expression Profiling of 16 Target Genes in Neoplastic and Non-Neoplastic Canine Mammary Tissues Using Branched-DNA Assay

Int J Mol Sci. 2016 Sep 21;17(9):1589. doi: 10.3390/ijms17091589.

Authors

Florenza Lüder Ripoli^{1

2}, Susanne Conradine Hammer^{3

4}, Annika Mohr^{5

6}, Saskia Willenbrock⁷, Marion Hewicker-Trautwein⁸, Bertram Brenig⁹, Hugo Murua Escobar¹⁰, Ingo Nolte¹¹

Affiliations

¹ Small Animal Clinic, University of Veterinary Medicine Hannover, Hannover D-30559, Germany. florenza@ripoli.com.br.
² Hematology Oncology and Palliative Medicine, Clinic III, University of Rostock, Rostock D-18057, Germany. florenza@ripoli.com.br.
³ Small Animal Clinic, University of Veterinary Medicine Hannover, Hannover D-30559, Germany. Susanne.Conradine.Hammer@tiho-hannover.de.
⁴ Hematology Oncology and Palliative Medicine, Clinic III, University of Rostock, Rostock D-18057, Germany. Susanne.Conradine.Hammer@tiho-hannover.de.
⁵ Small Animal Clinic, University of Veterinary Medicine Hannover, Hannover D-30559, Germany. annika.mohr@tiho-hannover.de.
⁶ Hematology Oncology and Palliative Medicine, Clinic III, University of Rostock, Rostock D-18057, Germany. annika.mohr@tiho-hannover.de.
⁷ Small Animal Clinic, University of Veterinary Medicine Hannover, Hannover D-30559, Germany. willenbrock@preclinics.com.
⁸ Department of Pathology, University of Veterinary Medicine Hannover, Hannover D-30559, Germany. marion.hewicker-trautwein@tiho-hannover.de.
⁹ Institute of Veterinary Medicine, Georg-August-University Göttingen, Göttingen D-37077, Germany. bbrenig@gwdg.de.
¹⁰ Hematology Oncology and Palliative Medicine, Clinic III, University of Rostock, Rostock D-18057, Germany. hugo.murua.escobar@med.uni-rostock.de.
¹¹ Small Animal Clinic, University of Veterinary Medicine Hannover, Hannover D-30559, Germany. ingo.nolte@tiho-hannover.de.

Abstract

Mammary gland tumors are one of the most common neoplasms in female dogs, and certain breeds are prone to develop the disease. The use of biomarkers in canines is still restricted to research purposes. Therefore, the necessity to analyze gene profiles in different mammary entities in large sample sets is evident in order to evaluate the strength of potential markers serving as future prognostic factors. The aim of the present study was to analyze the gene expression of 16 target genes (BRCA1, BRCA2, FOXO3, GATA4, HER2, HMGA1, HMGA2, HMGB1, MAPK1, MAPK3, MCL1, MYC, PFDN5, PIK3CA, PTEN, and TP53) known to be involved in human and canine mammary neoplasm development. Expression was analyzed in 111 fresh frozen (FF) and in 170 formalin-fixed, paraffin-embedded (FFPE) specimens of neoplastic and non-neoplastic canine mammary tissues using a multiplexed branched-DNA (b-DNA) assay. TP53, FOXO3, PTEN, and PFDN5 expression revealed consistent results with significant low expression in malignant tumors. The possibility of utilizing them as predictive factors as well as for assisting in the choice of an adequate gene therapy may help in the development of new and improved approaches in canine mammary tumors.

Keywords: RNA; canine mammary tumor; formalin-fixed; fresh frozen tissue; gene expression; multiplexed branched-DNA (b-DNA) assay; paraffin-embedded samples.