Low-Grade Mammary Gland Tumors in Dogs Have Greater VEGF-A and BMP2 Immunostaining and Higher CD31 Blood Vessel Density

Abstract

Tumor angiogenesis is an important process in tumor growth, and different molecules are involved in its regulation including VEGF-A, BMP2, and CD31, which can be considered possible prognostic markers. The aim of this study was to verify whether the VEGF-A and BMP2 immunostaining area, and microvascular density (MVD) might be associated with the degree of malignancy in malignant mammary neoplasms of dogs. For this purpose, samples of mammary malignancies from female dogs embedded in wax were used and separated into 4 main histomorphological types: tubulopapillary carcinomas, solid, complex, and carcinosarcoma, which were separated based on high and low degrees of malignancy. Immunohistochemical analysis was performed on tissue microarray blocks using anti-CD31 antibodies for evaluation of MVD and vascular lumen area, and with anti-VEGF-A and anti-BMP2 to determine the immunostaining area using the DAKO EnVision FLEX+ kit. MVD and vascular lumen area were higher in tubulopapillary carcinomas as were the areas stained by VEGF-A and BMP2. Immunostaining for CD31 was higher in low-grade carcinomas as well as in areas immunostained by VEGF-A and BMP2. There was a positive correlation between VEGF and BMP2 in high (r = 0.556, P < .0001) and low-grade (r = 0.287, P < .0001) carcinomas and between MVD and VEGF-A in low-grade carcinomas (r = 0.267, P = .0064). Thus, the markers evaluated showed greater immunostaining in canine mammary tumors with a lower degree of malignancy.