Relaionship among Expression of ESM-1, VEGF-D, Lymphangiogenesis and Lymph Node Metastasis in Human Invasive Lobular Breast Cancer
|Keywords||Invasive lobular breast cancer Endothelial cell specific molecule-1(ESM-1) Vascular endothelial growth factor-D Lymphatic microvessel density Lymphangiogenesis|
Background and objectiveMetastasis remains the most deadly aspect of breast carcinoma. In patients with breastcancer, lymph node metastasis is recognized as the most important prognostic indicator. Withthe discovery of specific lymphatic endothelial cells markers, such as D2-40, many studies havedemonstrated the presence of increasing newly formed lymphatic vessels in tumor tissue, whichis significantly correlated with the invasion of tumours, lymphatic metastasis, prognosis and soon. However, the reasons, process and molecular mechanisms of lymphangiogenesis inneoplasms are still unknown. Vascular endothelial growth factors -C, D are recognized as amajor lymphangiogenesis factor, can induce lymphangiogenesis by binding to thereceptor(VEGFR-3), the same as tumor angiogenesis, tumor lymphangiogenesis also needmultiple tumor-derived growth factor interactions. Recently, a literature reported that VEGFR-3‐mediated signaling can increase the expression of ESM-1, while ESM-1 also plays a centralrole in VEGFR‐ 3‐ mediated lymphangiogenesis in vitro and in experimental models.However, there is little research on the relationship between ESM-1 and VEGFR-3 axis inhuman solid tumour, especially the relationship among lymphangiogenesis, lymph metastasisand expression of ESM-1, VEGF-D in human breast carcinoma have not been reported yet.In the present study, we used immunohistochemistry to evaluate the expression pattern ofESM-1, VEGF-D and lymphatic microvessel density(using D2-40) in 108 infiltrating ductalcarcinoma (IDC) and 20 breast fibroadenoma in relation to the well known clinicopathologicalparameters, as well as other biological indicators of tumor behavior, such as c-erbB-2, ER andPR. We also detect the correlation between VEGF-D and ESM-1 expression infiltrating ductalcarcinoma (IDC). This may provide clues to reveal the mechanism of lymphangiogenesis, and italso provide better ways to prevent lymph metastasis in in infiltrating ductal carcinoma.Materials and methods(1) Our study assessed the cases of 108 Chinese infiltrating ductal carcinoma patients aged 29–77 years (mean age 44 years) who had been surgically treated, from January 2006 toJanuary 2008, in the Department of General Surgery, Second Affiliated Hospital, ShantouUniversity Medical College, Shantou, China, and all were deidentified prior to the study. All ofthese cases had no family history of breast cancer. We collected full clinical and pathologicaldata for these108 patients. The patients were all underwent curative surgery, and none of themhad received radiation or chemotherapy preoperation. We used a histories of another 20 patientswith breast fibroadenoma as controls. Lastly, an informed consent was obtained from patients inorder the material derived from them to be used in our research.(2) The expression of D2-40, ESM-1、VEGF-D in infiltrating ductal carcinoma were detectedby SP immunohistochemistrical method, and lymphatic microvessel density (LMVD) wasdetermined by lymphatic endothelial specific marker D2-40.(3) All of statistical analysis was performed with statistical package SPSS version 16.0software.Results(1) A positive immunoreaction of ESM-1 was observed, predominantly in cytoplasm andnuclei of the malignant cells, but not in normal breast epithelium. Vessel endothelial cells at theleading edge of the tumor were occasionally immunopositive for ESM-1 The positive rates ofESM-1 in infiltrating ductal carcinoma were significantly higher than that in the controlgroup(74.7.3% vs 10.0%, P < 0.01).(2) VEGF-D protein was immunodetected both in malignant cells and normal breast epithelium.The positive rates of VEGF-D in infiltrating ductal carcinoma were significantly higher thanthat in the control group（64.8% vs 15.0%, P < 0.01).(3) ESM-1 was related to larger tumor size and Lymph Nodes metastasis. No associations werefound between ESM-1 expression and other clinical parameters such as age, TNM stage, ER,PR and G-erbB2.(4) VEGF-D was significantly associated with Lymph Nodes metastasis. No associations werefound between VEGF-D expression and other clinical parameters such as tumor size, age, TNM stage, ER, PR and G-erbB2.(5) At the leading edge infiltrating ductal carcinoma Intensive lymphatic vessels showed larger,thin-walled structure and tumor embolus in their lumen. Lymphatic microvessels of humaninvasive lobular breast cancer show funicular or no lumen structure in intratumoral tissues. TheLMVD in breast carcinoma was significantly higher than in control tissues（17.63±4.65 vs5.80±1.11，T =11.21 , P<0.01).(6) Using Spearman correlation analysis, there was a statistically significant relationshipbetween ESM-1 expression and VEGF-D expression(r=0.711, p<0.01). LMVD rose graduallywith increasing expression of ESM-1 and VEGF-D in breast carcinoma（p<0.01）.Conclusion(1) In infiltrating ductal carcinoma, ESM-1 participate in the processes of Lymphangiogenesis,Lymph Nodes metastasis metastasis and tumor proliferation.(2) In infiltrating ductal carcinoma, The expression of ESM-1 was associated with the VEGF-Dexpression. It seems logical to postulate that VEGF-D could also increase ESM-1 expression,depended on the activity of both VEGFR-2 and VEGFR-3. In our study, LMVD was associatedwith ESM-1. We postulate that ESM-1 played an important role in VEGF-D/VEGFR-3-mediated lymphangiogenesis in a paracrine positive feedback loop(3) ESM-1 played an important role in VEGFR-3-mediated lymphangiogenesis not only invitro and in animal model, but also in human solid tumour. More experimental evidence iscontinually appearing on the involvement of ESM-1 in the control of tumor progression such astumor cells proliferation，migration and immune evasion, angiogenesis and lymphangiogenesis。If ESM-1 could be used as a valid therapeutic target, we can treat cancer through multiple ways.