Open Access Highly Accessed Research article

The endogenous soluble VEGF receptor-2 isoform suppresses lymph node metastasis in a mouse immunocompetent mammary cancer model

Masa-Aki Shibata1*, Jayakrishna Ambati2, Eiko Shibata13, Romulo JC Albuquerque2, Junji Morimoto4, Yuko Ito1 and Yoshinori Otsuki1

Author Affiliations

1 Department of Anatomy and Cell Biology, Division of Life Sciences, Osaka Medical College, Osaka, Japan

2 Department of Ophthalmology and Visual Sciences and Physiology, University of Kentucky, Lexington, KY, USA

3 Laboratory for Drug Discovery Innovation, Department of Molecular Pharmacology, National Cerebral & Cardiovascular Center Research Institute, Osaka, Japan

4 Laboratory Animal Center, Osaka Medical College, Osaka, Japan

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BMC Medicine 2010, 8:69  doi:10.1186/1741-7015-8-69

Published: 3 November 2010



Cancer metastasis contributes significantly to cancer mortality and is facilitated by lymphangiogenesis and angiogenesis. A new splicing variant, endogenous soluble vascular endothelial growth factor receptor-2 (esVEGFR-2) that we recently identified is an endogenous selective inhibitor of lymphangiogenesis. To evaluate the antimetastatic potential of esVEGFR-2, gene therapy with vector expressing esVEGFR-2 (pesVEGFR-2) or endostatin (pEndo) as a positive control was conducted on murine metastatic mammary cancer.


Syngeneic inoculated metastatic mammary cancers received direct intratumoral injection of pesVEGFR-2, pEndo or pVec as control, once a week for six weeks. In vivo gene electrotransfer was performed on the tumors after each injection.


Deaths from metastasis were much lower in the pesVEGFR-2 and pEndo groups than in those of the pVec. Tumor volume was significantly lower in the pesVEGFR-2 and the pEndo groups throughout the study. Multiplicity of lymph node and lung metastatic nodules was significantly suppressed in the pesVEGFR-2 and pEndo groups. Moreover, the total number of overall metastasis including the other organs was also decreased in these groups. However, pesVEGFR-2 was not able to decrease the number of lungs, ovaries, kidneys and adrenals with metastasis as counted by unilateral or bilateral metastasis. The number of CD34+/Lyve-1- blood microvessels was significantly decreased in the pEndo group, while the number of CD34-/Lyve-1+ lymphatic vessels was significantly decreased in the pesVEGFR-2 and pEndo groups. In addition, a significant reduction in the number of dilated lymphatic vessels containing intraluminal cancer cells was observed in the pesVEGFR-2 and pEndo groups. Levels of apoptosis were significantly increased in the pEndo group, whereas the rates of cell proliferation were significantly decreased in the pesVEGFR-2 and pEndo groups.


Our data demonstrate that esVEGFR-2 can inhibit mainly lymph node metastasis. The antimetastatic activity of esVEGFR-2 may be of high clinical significance in the treatment of metastatic breast cancer because lymph node involvement is a most important prognostic factor in cancer patients.