Assessment of endothelial cell proliferation in primary breast carcinoma and its association with axillary lymph node status (original) (raw)
Related papers
Clinical & Experimental Metastasis, 2000
It is well established that the ability of a neoplasm to induce a blood supply from a pre-existing circulation (angiogenesis) is a major factor in tumour growth, invasion and metastasis. However, the angiogenic potential of metastases and their subsequent growth have not been extensively studied. The question arises: can metastatic clones induce the same level of angiogenesis as in the primary neoplasm they emanated from? In this study it is hypothesised that in the same patient the level of vascularity and angiogenesis is the same in both the primary invasive ductal carcinoma and in the axillary lymph node metastasis at the time of surgery, according to Kerbels theory of clonal-dominance. To directly address the hypothesis, morphological measures of the established blood/lymphatic circulation (vascularity) as well as estimates of angiogenesis (endothelial cell proliferation) were measured in primary tumours and directly compared to the same parameters in the corresponding lymph node metastasis in a case by case basis (n = 17). The results demonstrate varying associations between the level of vascularity and angiogenesis between matched individual tumours and their metastatic lymph nodal deposits. It is possible that either variations in the angiogenic characteristics of the metastasising clone or local or systemic promoters or inhibitors of angiogenesis influence tumour angiogenesis at the different sites.
Clinical Cancer Research, 2008
Purpose: The interaction between tumor cells, stroma, and endothelial cells is important for the dissemination of tumor cells. The aim of the present study is to examine vascularity in primary breast carcinomas and its prognostic significance and relationship with tumor cell dissemination. Experimental Design: A total of 498 invasive breast carcinomas were analyzed. Representative tumor sections were stained for CD34 and CD105, and vascularity was quantified by the Chalkley method. The relationship between Chalkley counts, vascular invasion, disseminated tumor cells (DTC) in the bone marrow, other clinicopathologic variables, and clinical outcome was evaluated. Results: High vascular grades determined by Chalkley counts were significantly associated with shorter distant disease^free survival and breast cancer^specific survival in all patients (P < 0.001, log-rank) and in node-negative patients not receiving adjuvant systemic therapy (P < 0.05). In multivariate analysis, both CD34 and CD105 Chalkley counts showed prognostic significance for distant disease^free survival (P = 0.014 and P = 0.026), whereas CD34 also showed prognostic significance for breast cancer^specific survival (P = 0.007). Vascular invasion and DTCs in the bone marrow showed independent prognostic significance. DTC did not discriminate survival for CD34 low Chalkley counts, whereas a very poor prognosis was observed for DTC-positive patients with high CD34 counts. In node-negative patients not receiving systemic chemotherapy, high CD34 and high CD105 counts in combination identified patients with unfavorable outcome, as opposed to all other CD34/CD105 combinations. Conclusions: Improved identification of risk groups could be obtained by adding CD34 and CD105 vascular analysis to DTC, vascular invasion, and other primary tumor factors. This may facilitate the selection of candidates for adjuvant systemic therapy.
Breast Cancer Research and Treatment, 1998
Case-control methodology was used to evaluate the significance of vascularity in small breast carcinomas with regard to the presence or absence of axillary lymph node metastases. Vascularity was assessed in 32 axillary node positive primary breast tumours (LN+ve) less than 2 cm in size and compared with 56 control axillary node negative primary tumours (LN−ve), which were matched for histological type and grade and tumour size. This study design employed computer-assisted video analysis (CAVA) to assess the total blood vessel perimeter (BVP), total blood vessel area (BVA), and total blood vessel density (BVD) throughout a tissue section that encompassed an entire cross section of the tumour and its immediate periphery. The BVA and BVD in these tumours were not significantly different between LN+ve and LN−ve groups. The LN−ve carcinomas had, on average, a significantly (P < 0.05) higher total BVP (3355 µm/mm 2) than LN+ve tumours (2771 µm/mm 2). 'Hot spot' areas were also independently assessed by two pathologists and the same areas measured by CAVA. A strong correlation (P < 0.001) between the two methods of assessment of BVD of the neovascular 'hot spots' was found; however, no association with axillary lymph node metastasis was found using either method of assessment. In conclusion, vascularity assessed by either blood vessel density or blood vessel size in primary invasive breast cancers less than 2 cm in diameter showed no association with axillary lymph node metastasis; in fact a negative association was found with total BVP of whole tumour sections and BVD in 'hot spots' using CAVA. Further, this study has established a computer-assisted method of quantifying vascularity in solid neoplasms and is a positive step towards a standardised approach to this diverse and methodologically variable area.
Examining the technique of angiogenesis assessment in invasive breast cancer
British Journal of Cancer, 1997
The intensity of angiogenesis as measured by the density of microvessels has been reported to be associated with a poor prognosis in invasive breast cancer in some, but not all, studies. The reasons for these discrepancies may be variations in the methodologies used. The monoclonal antibody used to identify the microvessels, the number of high-density areas or 'hotspots' counted and the type of value taken for statistical analysis (highest count or mean count) have varied between the different studies. We have assessed which of the three commonly used monoclonal antibodies provides the best visualization of microvessels in invasive breast cancer and have used methods that give reproducible data for the optimum number of 'hotspots' to count for each reagent. Thus, microvessels in formalin-fixed paraffin-embedded specimens from 174 primary breast cancers were immunohistochemically stained with monoclonal antibodies to FVIIIRAg, CD31 and CD34 and ten fields counted at 200 x magnification for each antibody. The highest count and the mean value of the highest of three, five and ten counts were used to examine the relationship between the density of microvessels and overall survival of patients with a median follow-up time of 7.1 years. Antibodies to CD31 and CD34 identified more vessels than antibodies to FVIIIRAg (median highest count per mm2: CD31 = 100, CD34 = 100, FVIIIRAg = 81). The monoclonal antibody to CD31, however, was the least reliable antibody, immunohistochemically staining only 87% of sections compared with 98% for the monoclonal to CD34 and 99% for the monoclonal to FVIIIRAg. There was a high degree of correlation between the number of vessels stained by the different antibodies, though there were some considerable differences in actual counts for serial sections of the same specimen stained by the different antibodies. Patients could be divided into two groups corresponding to those with high microvessel densities and those with low microvessel densities. Using Kaplan-Meier survival curves, there was a close association for all three antibodies between vessel density and survival whichever method of recording the highest vessel densities was used. Using log-rank tests and Cox's regression analysis, anti-CD34 gave the most significant results of the three antibodies, whereas a simple cutoff at the 75th percentile for the high and low groups produced the best association with patient survival. For anti-CD34 the highest microvessel density (P = 0.0014) and the mean value of the highest three microvessel densities (P= 0.004) showed a good correlation with patient death, whereas for anti-CD31 (P= 0.008) and anti-FVIIIRAg (P= 0.007) the highest count gave the best correlation using Cox's regression analysis.
Evaluation of Vascular Proliferation in Molecular Subtypes of Breast Cancer
in Vivo, 2018
Background: Angiogenesis plays a pivotal role in tumor development. Although microvessel density (MVD) is the most common method used for evaluation of angiogenesis, it has several limitations. Our aim was to evaluate MVD and microvessel proliferation (MVP) in a series of invasive breast carcinomas and analyze whether angiogenesis is influenced by the molecular phenotype of each tumor. Materials and Methods: We examined vascular proliferation using double immunohistochemistry (CD34/Ki67) in a series of 54 invasive breast carcinomas and compared the results with standard MVD, molecular subtypes and other classical parameters. Results: Increased MVD and MVP values were recorded in basal-like subtype, but only the MVP value reached significance among this group of patients (p=0.0001). For all cases combined, increased MVP was significantly correlated with negative estrogen receptor (ER) status (p=0.010) and higher histological grade (p=0.002). Conclusion: MVP more accurately reflects the state of angiogenesis in breast cancer, compared with standard MVD. Vascular proliferation was associated with aggressive tumor features, indicating its contribution to tumor progression. The strong association between vascular proliferation and basal-like tumors suggests that this marker can be used for stratification of patients who might benefit from therapies targeting angiogenesis. Angiogenesis is considered a hallmark of cancer and a key requisite in their growth, invasion and progression (1). In 1971, Folkman suggested that tumors can be treated by inhibiting their vascularization (2). It is well known that tumors cannot exceed 2-3 mm without vascular support (2), thus, anti-angiogenic therapy is an attractive target for angiogenesis-dependent tumors such as breast cancer. Microvessel density (MVD) is the most widely method used for evaluation of angiogenesis, based on counting the vessels in the most vascularized areas of the tumors, namely 'hot spots'. This method was developed by Weidner and coworkers in 1991, who demonstrated that MVD influences the prognostic of patients with breast cancer (3, 4). Since then, many other researchers have investigated the role of MVD in breast tumors, but the results are contradictory. However, MVD has some limitations, as it cannot predict the response to therapy or the treatment efficacy (5). Recent studies showed that microvessel proliferation (MVP), defined as the average number of vessels exhibiting co-expression of an endothelial and a proliferation marker, is a better indicator of angiogenesis compared with MVD (5-7). In prostate and endometrial carcinomas, microvessel proliferation was found to be a more reliable prognostic marker compared with standard MVD (5-7). With this background, the aim of the present study was to evaluate vascular proliferation (CD34/Ki67 co-expression) and standard MVD in a series of invasive breast carcinomas, in accordance with the molecular classification. The results were compared by classical clincopathological parameters. Materials and Methods The present study included 54 female patients, aged between 39-85 years (mean=57.3 years), who underwent radical modified mastectomy and lymph node dissection between 2009-2013. Surgical specimens were fixed in buffer formalin and paraffin embedded and 5 μm-thick step sections were performed for each 79 This article is freely accessible online.
Assessment of Lymphatic Vessel Density in Breast Carcinoma Using Immunohistochemical Marker D 2-40
2018
Background: Breast carcinoma is the most common malignant tumor in females and the incidence of this disease has been significantly increased. Metastasis is the leading cause of mortality in patients diagnosed with breast cancer. It relies heavily on development of new blood vessels (angiogenesis) and lymphatics (lymphangiogenesis) Objective: The aim of the present study was to assess the significance of lymphatic vessel quantitation in breast carcinoma. Methods: The study included 50 cases of invasive breast cancer. Lymphatic microvessels were identified by using immunohistochemical stain D2-40 in tumoral and peritumoral area. The microvessels were counted within a 400 x magnification field in the area of highest microvessel density. Result: -Intratumoral lymphatic vessel density and peritumoral lymphatic vessel density in node positive cases was significantly correlated. Conclusion: A significant intra and peritumoral lymphatics vessel density in node positive patients indicate th...
Breast Cancer Research and Treatment, 2009
Controversy exists regarding the topography of lymph vessels in breast cancer, their usefulness as prognostic factors, relationship with angiogenesis and whether active lymphangiogenesis occurs within the tumour. A series of 177 well-characterized breast cancers, with long term follow up, were stained with D2-40, CD31 and CD34. Distribution of lymphatics and lymph vessel density (LVD) were assessed in three areas, intratumoural, peripheral and peritumoural and correlated with clinicopathological criteria and patient prognosis. Microvessel density (MVD) was assessed and correlated with LVD. Double immunohistochemical staining with D2-40 and MIB-1 was carried out to assess the proliferative status of lymphatics and of the tumour emboli within. Peritumoural lymphatics were detected in all tumours whereas peripheral and intratumoural lymphatics were detected in 86 and 41% of specimens, respectively. Tumours with higher total LVD were significantly associated with the presence of lymph node (LN) metastasis and shorter overall survival (OS). In multivariate analysis, tumour grade, LN status and the presence of lymphovascular invasion, but not LVD, were independent poor prognostic factors. No association was found between LVD and MVD. Proliferating lymphatics were detected in 29% of specimens and were significantly associated with dense inflammatory infiltrate. In conclusion, lymphatics are located primarily in the peritumoural and peripheral areas in breast cancer and seem to play an important role in disease progression by being routes for tumour dissemination. The lack of correlation between lymphangiogenic and angiogenic characteristics suggests two distinct processes and the presence of active lymphangiogenesis, albeit in a small portion of specimens, may have important therapeutic implications.
Angiogenesis in breast cancers without lymph node metastasis
Turkish Journal of Pathology, 2010
Objective: Many studies have been focused on angiogenesis as a possible prognostic factor, recently. In this study, we evaluated microvessel density by CD34 in primary tumors of the patients with lymph node negative breast carcinomas and compared the results with prognostic parameters. Material and Method: 31 patients with invasive breast carcinomas without axillary lymph node metastasis were included in this study. Microvessel density was assessed by CD34 staining in the primary tumor and compared with the tumor type and prognostic parameters such as age, tumor size, nuclear grade, lymphatic vessel invasion, and estrogen and progesteron reseptor status. CD34 staining was evaluated in three areas by x200 magnification. Results: The mean value microvessel density of CD34 staining in the primary tumor was 43 (minimum 19 and maximum 100). There was a significant relationship between microvessel density and progesterone receptor expression. microvessel density was higher in lobular type breast carcinoma. Conclusion: The correlation between progesterone receptor status and microvessel density should be further confirmed by prospective studies. The high microvessel density rate in lobular carcinoma may be attributed to abundance of stromal cells or to the low number of patients.