Distinguishing blood and lymph vessel invasion in breast cancer: a prospective immunohistochemical study

Abstract

Recently, peritumoural (lympho)vascular invasion, assessed on haematoxylin -eosin (HE)-stained slides, was added to the St Gallen criteria for adjuvant treatment of patients with operable breast cancer (BC). New lymphatic endothelium-specific markers, such as D2-40, make it possible to distinguish between blood (BVI) and lymph vessel invasion (LVI). The aim of this prospective study was to quantify and compare BVI and LVI in a consecutive series of patients with BC. Three consecutive sections of all formalin-fixed paraffinembedded tissue blocks of 95 BC resection specimens were (immuno)histochemically stained in a fixed order: HE, anti-CD34 (panendothelium) and anti-D2-40 (lymphatic endothelium) antibodies. All vessels with vascular invasion were marked and relocated on the corresponding slides. Vascular invasion was assigned LVI (CD34" or~/D2-40") or BVI (CD34"/D2-40~) and intra-(contact with tumour cells or desmoplastic stroma) or peritumoural. The number of vessels with LVI and BVI as well as the number of tumour cells per embolus were counted. Results were correlated with clinico-pathological variables. Sixty-six (69.5%) and 36 (37.9%) patients had, respectively, LVI and BVI. The presence of 'vascular' invasion was missed on HE in 20% (peritumourally) and 65% (intratumourally) of cases. Although LVI and BVI were associated intratumourally (P ¼ 0.02), only peritumoural LVI, and not BVI, was associated with the presence of lymph node (LN) metastases (p peri ¼ 0.002). In multivariate analysis, peritumoural LVI was the only independent determinant of LN metastases. Furthermore, the number of vessels with LVI was larger than the number of vessels with BVI (P ¼ 0.001) and lymphatic emboli were larger than blood vessel emboli (P ¼ 0.004). We demonstrate that it is possible to distinguish between BVI and LVI in BC specimens using specific lymphatic endothelium markers. This is important to study the contribution of both processes to BC metastasis. Furthermore, immunohistochemical detection of lymphovascular invasion might be of value in clinical practice.

Significance of lymph vessel invasion identified by the endothelial lymphatic marker D2-40 in node negative breast cancer

“…34,35 The distinction between lymph vessel and vascular invasion in breast cancer has also been demonstrated using D2-40 and the vascular marker CD34. 36 Furthermore D2-40 has been shown to be the best marker for lymphatic endothelium. 37 In this study, we identified lymph vessel invasion using D2-40 in 82/303 (27%) of cases of node negative breast cancer.…”

“…Van Eynden's study using D2-40 also showed that lymphovascular invasion was missed on H&E in 20%(peritumorally) and in 65% (intratumorally) of breast cancer cases. 36 The subgroup analysis of the 46 cases whose tumors were H&EÀ/D2-40 þ and the 36 cases that Endothelial lymphatic marker D2-40 A Arnaout-Alkarain et al…”

“…Other studies have also reported that vascular invasion is less frequent than lymphatic invasion in breast cancer. 7,36 As only 17 tumors were H&E þ /D2-40À compared to the 204 cases that were H&EÀ/D2-40-, subgroup analysis could not be performed owing to the small numbers of H&E þ /D2-40À cases. Nevertheless, D2-40 could be useful in distinguishing true lymphatic invasion from retraction artefact.…”

“…34,35 In a pilot study of 50 breast cancers, D2-40 increased the detection of lymph vessel invasion by 20% in node-negative cases and 16% in node-positive cases. 34 Van den Eynden et al's 36 study also showed that D2-40 distinguished lymphatic invasion from blood vessel invasion in breast cancers. In the present study, we correlated the presence of lymph vessel invasion identified by D2-40 with the clinicopathological factors in primary node-negative breast cancers.…”

Podoplanin is a useful diagnostic marker for epithelioid hemangioendothelioma of the liver

“…2 Distinguishing between lymphatic vessels and blood capillaries is difficult using only hematoxylin and eosin (H&E) staining, and a commercially available D2-40 antibody reactive to podoplanin has been used to evaluate the lymphatic invasion of cancer in pathological examinations. 3 In addition, podoplanin was reported to be a useful marker for the diagnosis of vascular and non-vascular tumors of various organs. 4 Podoplanin was expressed by some vascular tumors with lymphatic differentiation, such as lymphangioma, Kaposi's sarcoma, Kaposiform hemangioendothelioma (socalled Dabska tumor), hobnail hemangioma, and a subset of angiosarcomas.…”