Lymphatic and blood vessel morphometry in invasive breast carcinomas: relation with proliferation and VEGF-C and -D proteins expression.

INTRODUCTION The aim of the present study was to investigate the distribution of both lymphatics and blood microvessels in invasive breast carcinomas and the clinicopathological and prognostic significance of their density and size related parameters as well as their correlation with the proliferative potential of the tumor and VEGF-C and -D expression. METHODS Both single and double immunohistochemistry were applied on a series of 146 paraffin-embedded breast tissue specimens to detect VEGF-C and -D as well as lymphatics and blood microvessels, respectively. Computer-assisted morphometry was performed to evaluate the blood and lymphatic vessel density (BVD and LVD respectively) as well as various vascular size related parameters. RESULTS Lymphatics were detected within the stroma at the tumor border, while blood vessels were located in both the interior of the tumor mass and peritumor stroma. BV major axis, minor axis and perimeter inversely correlated with ER (p=0.011, p=0.023 and p=0.008 respectively), while LV major axis, minor axis and the perimeter inversely correlated with tumor nuclear grade (p=0.045, p=0.037 and p=0.032 respectively) and topoisomerase IIalpha (p=0.015, p=0.024 and p=0.045 respectively). The same LV parameters were found to positively correlate with cancerous VEGF-C (p<0.0001, p=0.092 and p=0.012 respectively) and VEGF-D in the stromal fibroblasts surrounding neoplastic cells (p=0.011, p=0.041 and p=0.026 respectively). High BVD exerted an unfavorable impact on both disease-free (p=0.021) and overall survival (p=0.031) of the patients. High LVD correlated with poor disease-free and overall survival only in the subgroup of patients with ER-negative tumors (p=0.056 and p=0.0312 respectively). CONCLUSION These findings, for the first time, correlate lymphatic size with tumors of limited proliferative potential and higher nuclear differentiation. Moreover, they suggest that VEGF-C and -D expression influence lymphatic size rather than being involved in the increase of lymphatic vessel number.

[1]  S. Badve,et al.  Lymphangiogenesis Does Not Occur in Breast Cancer , 2005, The American journal of surgical pathology.

[2]  Misa Nakamura,et al.  Lymph vessel density correlates with nodal status, VEGF-C expression, and prognosis in breast cancer , 2005, Breast Cancer Research and Treatment.

[3]  P. Heikkilä,et al.  High LYVE-1–Positive Lymphatic Vessel Numbers Are Associated with Poor Outcome in Breast Cancer , 2004, Clinical Cancer Research.

[4]  D. Massi,et al.  Tumor lymphangiogenesis in head and neck squamous cell carcinoma , 2004, Cancer.

[5]  Michael Gnant,et al.  Prognostic Value of Lymphangiogenesis and Lymphovascular Invasion in Invasive Breast Cancer , 2004, Annals of surgery.

[6]  M. Karkkainen,et al.  Lymphatic vasculature: development, molecular regulation and role in tumor metastasis and inflammation. , 2004, Trends in immunology.

[7]  A. Shvarts,et al.  Lack of lymphangiogenesis during breast carcinogenesis , 2004, Journal of Clinical Pathology.

[8]  B. Uzzan,et al.  Microvessel density as a prognostic factor in women with breast cancer: a systematic review of the literature and meta-analysis. , 2004, Cancer research.

[9]  S. Tsutsui,et al.  Prognostic value of microvessel density in invasive ductal carcinoma of the breast , 2003, Breast cancer.

[10]  M. Tonato,et al.  Evaluation of the Prognostic Role of Vascular Endothelial Growth Factor and Microvessel Density in Stages I and II Breast Cancer Patients , 2003, Breast Cancer Research and Treatment.

[11]  A. Harris,et al.  Absence of lymphangiogenesis and intratumoural lymph vessels in human metastatic breast cancer , 2003, The Journal of pathology.

[12]  Alexander Marks,et al.  A New Monoclonal Antibody, D2-40, for Detection of Lymphatic Invasion in Primary Tumors , 2002, Laboratory Investigation.

[13]  S. Stacker,et al.  The role of tumor lymphangiogenesis in metastatic spread , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[14]  P. Carmeliet,et al.  Molecular mechanisms of lymphangiogenesis in health and disease. , 2002, Cancer cell.

[15]  T. Veikkola,et al.  Signalling via vascular endothelial growth factor receptor‐3 is sufficient for lymphangiogenesis in transgenic mice , 2001, The EMBO journal.

[16]  M. Pepper,et al.  Lymphangiogenesis and tumor metastasis: myth or reality? , 2001, Clinical cancer research : an official journal of the American Association for Cancer Research.

[17]  Steven A. Stacker,et al.  VEGF-D promotes the metastatic spread of tumor cells via the lymphatics , 2001, Nature Medicine.

[18]  R. Jain,et al.  Absence of functional lymphatics within a murine sarcoma: a molecular and functional evaluation. , 2000, Cancer research.

[19]  P. Athanassiadou,et al.  DNA Topoisomerase II-Alpha Immunoreactivity as a Marker of Tumor Aggressiveness in Invasive Breast Cancer , 2000, Pathobiology.

[20]  M. Oda,et al.  Increased vascular endothelial growth factor and vascular endothelial growth factor-c and decreased nm23 expression associated with microdissemination in the lymph nodes in stage I non-small cell lung cancer. , 2000, The Journal of thoracic and cardiovascular surgery.

[21]  K. Alitalo,et al.  Vascular endothelial growth factor D (VEGF-D) is a ligand for the tyrosine kinases VEGF receptor 2 (Flk1) and VEGF receptor 3 (Flt4). , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Yihai Cao,et al.  Proteolytic processing regulates receptor specificity and activity of VEGF‐C , 1997, The EMBO journal.

[23]  S. Pinder,et al.  Histological grading of breast carcinomas: a study of interobserver agreement. , 1995, Human pathology.

[24]  J. Folkman,et al.  Tumor angiogenesis and metastasis--correlation in invasive breast carcinoma. , 1991, The New England journal of medicine.

[25]  S. Fox,et al.  More about: Tumor angiogenesis as a prognostic assay for invasive ductal breast carcinoma. , 2000, Journal of the National Cancer Institute.

[26]  J. Folkman,et al.  Angiogenesis and angiogenesis inhibition: an overview. , 1997, EXS.