Expression of Lymphangiogenesis Markers and Correlation with Lymph Nodal Metastasis in Breast cancer
Authors: Vuthaluru Srineil , Mathur Sandeep , Seenu  , Srivastava Anurag 
Published: June 2021

Aim

To study expression of VEGF and D II40 in breast cancer and correlate with lymph nodal metastasis.

Materials & Methods

Formalin fixed paraffin embedded blocks of breast cancer were retrieved and 4 mu thick sections obtained& stained with routine ‘H&E’. Sections were cut on APES (3‐Triethoxysilylpropylamine) coated slides & immunohistochemistry (IHC) was performed using Streptavidin Biotin Peroxidase technique using monoclonal antibodies against VEGF and DII 40 on coated sections of cases & controls. IHC stains were reviewed in conjunction with H&E. Intensity of VEGF expression & its distribution in the tumour were observed under high power magnification & semi‐quantitatively analyzed. Brown granules in the cytoplasm/membrane of the cells were considered as +ve for VEGF expression when the proportion of immunoreactive cells was ≥5%. ; 5–25% staining, weakly +ve (+) 26–50% staining,+ve (++); >50% staining, strongly +ve (+++). For statistical analysis, samples with 0/+ staining considered in low expression group, while ++/+++ expression were considered as high expression. For DII40 well demarcated vessels showing positivity in endothelial cells were counted per 10 hpf after selecting hot spots at scanner view. For statistical analysis, 0 was considered as no expression and 1/10HPF was considered as high expression.

Results

48 tissue specimens were included in the study. High VEGF expression (Fig 1) was seen 33/43 patients (76.74%%; 95% CI 61.37%–88.24%) and high DII 40 expression (Fig 2) in 30/44 (68.18%; 95% CI 52.42%–81.39%). Variation in expression of VEGF and DII 40 in relation to each other is given in figure 1. Of the 33 patients with high VEGF expression, DII 40 was expressed in 26 of them (78.79%). There is a strong association between VEGF expression and DII 40 expression (p<0.01). The scatter diagram of VEGF and DII 40 expression showed a significant trend (Figure 3 ) indicating that as VEGF expression increases so does the DII 40 expression. The variance explained by VEGF level in the variance of DII 40 as measured by the correlation (R2) was 0.1265 (p=0.02). When VEGF expression was compared with LN status, low expression of VEGF was found in 38.47 % (5/13) of SLN positive and 16.67 % (5/30) of negative senitnel node patients. High expression of VEGF was in 61.53% of positive patients as compared to 83.33% of negative sentinel node patients. Results of VEGF expression and sentinel node status revealed no statistically significant relation between expression of VEGF and LN involvement. No D II 40 expression of was found in 38.46% (5/13) % of LN +ve patients as compared to 29.03% (9/31) in node negative. High D II 40 expression was found 61.54% LN+ve patients and 70.97% of snode negative. Results of DII 40 expression and LN status revealed no statistically significant relation between expression of DII 40 and node involvement. When the expression of VEGF and D II 40 was compared with tumour characteristics there was no statistically significant relation with tumour size, tumour type, grade, ER/PR staus and Her2neu status.

Conclusions

Lympahngiogenesis marker (VEGF) expression is high in most tumours and there is significant association between lymph vessels detection as studied by D II 40 expression and lymphangiogenesis marker expression as studied VEGF expresssion. However, no association was found between expression of VEGF and D II 40 with neither sentinel / non‐sentinel nodal status nor any of the primary tumour characteristics.

Support or Funding Information

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This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.