Angiogenesis and lymphangiogenesis are essential for
breast cancer progression and are regulated by
vascular endothelial growth factors (
VEGF). To determine clinical and molecular correlates of these processes, we measured blood and lymphatic vascular microvessel density in 29 invasive
carcinomas (22 ductal, six lobular, one papillary), using the vascular marker CD31 and the novel lymphatic marker D2-40. Microvessel density was assessed microscopically and by image cytometry, and was compared with
tumor histology, grade, stage,
lymph node metastasis,
hormone receptors, HER2/neu status, and expression of
VEGF,
VEGF-C and
VEGF-D by immunohistochemistry or quantitative RT-PCR. Strong correlation was observed between visual and image cytometric microvessel density using D2-40 but not CD31 (P=0.016 and 0.1521, respectively). Image cytometric CD31 microvessel density correlated with
tumor size, grade, stage and
lymph node metastasis (P=0.0001, 0.0107, 0.0035 and 0.0395, respectively). D2-40 microvessel density correlated with
tumor stage (P=0.0123 by image cytometry) and
lymph node metastasis (P=0.0558 by microscopy). Immunohistochemical
VEGF signal in peritumoral blood vessels correlated with image cytometric CD31 and D2-40 microvessel density (P=0.022 and 0.0012, respectively), consistent with the role of
VEGF in blood and lymphatic vascular growth. Intratumoral
VEGF-C and
VEGF-D expression by quantitative RT-PCR correlated with D2-40 (P=0.0291 by image cytometry) but not with CD31 microvessel density, which could suggest a selective role of
VEGF-C and
VEGF-D in lymphangiogenesis. CD31 and D2-40 microvessel density correlated significantly with several prognostic factors, including
lymph node metastasis. Thus, measurements of angiogenesis and lymphangiogenesis may have utility for
breast cancer pathology, particularly for estimation of metastatic risk.