Lymphangiogenesis as a Prognostic Marker in Breast Cancer Using D2-40 as Lymphatic Endothelial Marker—A Preliminary Study

Mumtaz A. Ansari, Vaibhav Pandey, Vivek Srivastava, Mohan Kumar, R. N. Mishra, Anand Kumar

Department of Biostatistics, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India.
Department of General Surgery, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India.
Department of Pathology, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India.

DOI: 10.4236/jct.2012.325103                

Abstract

We studied tumour lymphangiogenesis and lymphatic invasion using D2-40 endothelial marker in 35 breast cancer patients treated by primary surgery and correlated it with various clinico-pathological prognostic parameters. Lymphangiogenesis was quantified using lymphatic micro vessel density (LMVD) by counting the immunostained lymphatic microvessels at 200X. The mean age was 45.97±12.09 years (range 30-80 years). LMVD ranged from 5/hpf to 56/hpf with a mean score of 13.4±10.8 and median of 9. The median value of 9 was taken to classify patients into a low or high LMVD. LMVD correlated significantly with tumour size (p=0.003), histological grade (p=0.046), lymph node status (p=0.030). There was no significant correlation of LMVD with stage, estrogen receptor, progesterone receptor or HER2/neu immunoreactivity. Lymphovascular invasion on D2-40 staining [LVI-D40] was found in 13 (37.1%) cases compared to 6 cases (17.1%) on H & E staining showing a poor agreement (k=0.244). LVI correlated significantly with lymph node status (p=0.011). There was a strong association between tumour size (p=0.142), histological grade (p=0.066) though the correlation was not statistically significant. No correlation was found with stage, estrogen receptor, progesterone receptor or HER2/neu immunoreactivity. The mean LMVD in LVI positive patients was higher (22.85±13.29) as compared to LVI negative patients (7.95±2.05) and this was statistically significant (p=0.001). Increased D2-40 detected LMVD and LVI correlated with poor prognostic parameters.

Keywords

Breast Cancer; Lymphangiogenesis, D2-40; Prognosis

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	E. R. Fisher, et al., “Pathologic findings from the National Surgical Adjuvant Breast Project (NSABP) Protocol B-17: Intraductal Carcinoma (Ductal Carcinoma in Situ),” Cancer, Vol. 75, No. 6, 1995, pp. 1310-1319. doi:10.1002/1097-0142(19950315)75:6<1310::AID-CNCR2820750613>3.0.CO;2-G
[2]	P. A. Kyzas, et al., “Evidence for Lymphangiogenesis and Its Prognostic Implications in Head and Neck Squamous Cell Carcinoma,” Journal of Pathology, Vol. 206, No. 2, 2005, pp. 170-177. doi:10.1002/path.1776
[3]	K. Schmid, et al., “Prognostic Value of Lymphatic and Blood Vessel Invasion in Neuroendocrine Tumours of the Lung,” The American Journal of Surgical Pathology, Vol. 29, 2005, pp. 324-328. doi:10.1097/01.pas.0000149706.74216.b6
[4]	P. Bono, et al., “High LYVE-1-Positive Lymphatic Vessel Numbers Are Associated with Poor Outcome in Breast Cancer,” Clinical Cancer Research, Vol. 10, No. 21, 2004, pp. 7144-7149. doi:10.1158/1078-0432.CCR-03-0826
[5]	C. S. M. Williams, et al., “Absence of Lymphangiogenesis and Intratumoural Lymph Vessels in Human Metastatic Breast Cancer,” The Journal of Pathology, Vol. 200, 2003, pp. 195-206. doi:10.1002/path.1343
[6]	H. J. Kahn, et al., “Monoclonal Antibody D2-40, a New Marker of Lymphatic Endothelium, Reacts with Kaposi’s Sarcoma and a Subset of Angiosarcomas,” Modern Pathology, Vol. 15, No. 4, 2002, pp. 434-440. doi:10.1038/modpathol.3880543
[7]	N. Weidner, et al., “Tumour Angiogenesis and Metastasis—Correlation in Invasive Breast Carcinoma,” The New England Journal of Medicine, Vol. 324, No. 1, 1991, pp. 1-8. doi:10.1056/NEJM199101033240101
[8]	R. G. C. Dumitrescu, “Understanding Breast Cancer Risk Where Do We Stand in 2005?” Journal of Cellular and Molecular Medicine, Vol. 9, No. 1, 2005, pp. 208-221. doi:10.1111/j.1582-4934.2005.tb00350.x
[9]	A. J. Leu, et al., “Absence of Functional Lymphatics within a Murine Sarcoma: A Molecular and Functional Evaluation,” Cancer Research, Vol. 60, No. 16, 2000, pp. 4324-4327.
[10]	M. Skobe, et al., “Induction of Tumour Lymphangiogenesis by VEGF-C Promotes Breast Cancer Metastasis,” Nature Medicine, Vol. 7, No. 16, 2001, pp. 192-198. doi:10.1038/84643
[11]	S. J. Mandriota, et al., “Vascular Endothelial Growth Factor-C Mediated Lymphangiogenesis Promotes Tumour Metastasis,” The EMBO Journal, Vol. 72, No. 20, 2001, pp. 672-682. doi:10.1093/emboj/20.4.672
[12]	Y. He, et al., “Suppression of Tumour Lymphangiogenesis and Lymph Node Metastasis by Blocking Vascular Endothelial Growth Factor Receptor 3 Signaling,” Journal of the National Cancer Institute, Vol. 94, No. 11, 2002, pp. 819-825. doi:10.1093/jnci/94.11.819
[13]	S. Breiteneder-Geleff, et al., “Angiosarcomas Express Mixed Endothelial Phenotypes of Blood and Lymphatic Capillaries: Podoplanin as a Specific Marker for Lymphatic Endothelium,” American Journal of Pathology, Vol. 154, 1999, pp. 385-394. doi:10.1016/S0002-9440(10)65285-6
[14]	M. Schmelz and W. W. Franke, “Complexus Adhaerentes, a New Group of Desmoplakin-Containing Junctions in Endothelial Cells: The Syndesmos Connecting Retothelial Cells of Lymph Nodes,” European Journal of Cell Biology, Vol. 61, No. 2, 2005, pp. 274-289.
[15]	J. T. Wigle and G. Oliver, “Prox1 Function Is Required for the Development of the Murine Lymphatic System,” Cell, Vol. 98, No. 2, 1999, pp. 769-778. doi:10.1016/S0092-8674(00)81511-1
[16]	D. Massi, et al., “Tumour Lymphangiogenesis Is a Possible Predictor of Sentinel Lymph Node Status in Cutaneous Melanoma: A Case-Control Study,” Journal of Clinical Pathology, Vol. 59, No. 2, 2006, pp. 166-173. doi:10.1136/jcp.2005.028431
[17]	Y. Miyata, et al., “Lymphangiogenesis and Angiogenesis in Bladder Cancer: Prognostic Implications and Regulation by Vascular Endothelial Growth Factors-A, -C, and -D,” Clinical Cancer Research, Vol. 12, 2006, pp. 800-806. doi:10.1158/1078-0432.CCR-05-1284
[18]	H. J. Kahn and A. Marks, “A New Monoclonal Antibody, D2-40, for Detection of Lymphatic Invasion in Primary Tumours,” Laboratory Investigation, Vol. 82, 2002, pp. 1255-1257.
[19]	Marinho VF, et al., “Lymph Vascular Invasion in Invasive Mammary Carcinomas Identified by the Endothelial Lymphatic Marker D2-40 Is Associated with Other Indicators of Poor Prognosis,” BMC Cancer, Vol. 8, 2008, pp. 64-68. doi:10.1186/1471-2407-8-64
[20]	S. F. Schoppmann, et al., “Prognostic Value of Lymphangiogenesis and Iymphovascular Invasion in Invasive Breart Cancer,” Annals of Surgery, Vol. 240, No. 2, 2004, pp. 306-312. doi:10.1097/01.sla.0000133355.48672.22
[21]	X. D. Xie, et al., “Relation of Lymphatic Microvessel Density Detected by Monoclonal Antibody D2-40 with VEGF-C Expression in Breast Cancer,” Zhonghua Zhong Liu Za Zhi, Vol. 30, No. 5, 2008, pp. 356-360.
[22]	S. D. Nathanson, “Insights into the Mechanisms of Lymph Node Metastasis,” Cancer, Vol. 98, No. 2, 2003, pp. 413-423. doi:10.1002/cncr.11464
[23]	G. Gurleyik, et al., “Lymphovascular Invasion, as a Prognostic Marker in Patients with Invasive Breast Cancer,” Acta Chirurgica Belgica, Vol. 107, No. 3, 2007, pp. 284-287.
[24]	R. Jagsi, et al., “Locoregional Recurrence Rates and Prognostic Factors for Failure in Node-Negative Patients Treated with Mastectomy: Implications for Postmastectomy Radiation,” International Journal of Radiation Oncology, Biology, Physics, Vol. 62, No. 4, 2005, pp. 1035-1039. doi:10.1016/j.ijrobp.2004.12.014
[25]	B. Kuru, et al., “Prognostic Factors Affecting Survival and Disease-Free Survival in Lymph Nodenegative Breast Carcinomas,” Journal of Surgical Oncology, Vol. 83, 2003, pp. 167-172. doi:10.1002/jso.10264
[26]	S. D. Nathanson, et al., “Sentinel Lymph Node Metastasis in Experimental Melanoma: Relationships among Primary Tumour Size, Lymphatic Vessel Diameter and 99mTclabeled Human Serum Albumin Clearance,” Annals of Surgical Oncology, Vol. 4, No. 2, 1997, pp. 161-168. doi:10.1007/BF02303800