Abstract
The neurotrophic factor ARTEMIN (ARTN) has been reported to possess a role in mammary carcinoma progression and metastasis. Herein, we report that ARTN modulates endothelial cell behaviour and promotes angiogenesis in ER-mammary carcinoma (ER-MC). Human microvascular endothelial cells (HMEC-1) do not express ARTN but respond to exogenously added, and paracrine ARTN secreted by ER-MC cells. ARTN promoted endothelial cell proliferation, migration, invasion and 3D matrigel tube formation. Angiogenic behaviour promoted by ARTN secreted by ER-MC cells was mediated by AKT with resultant increased TWIST1 and subsequently VEGF-A expression. In a patient cohort of ER-MC, ARTN positively correlated with VEGF-A expression as measured by Spearman's rank correlation analysis. In xenograft experiments, ER-MC cells with forced expression of ARTN produced tumors with increased VEGF-A expression and increased microvessel density (CD31 and CD34) compared to tumors formed by control cells. Functional inhibition of ARTN by siRNA decreased the angiogenic effects of ER-MC cells. Bevacizumab (a humanized monoclonal anti-VEGF-A antibody) partially inhibited the ARTN mediated angiogenic effects of ER-MC cells and combined inhibition of ARTN and VEGF-A by the same resulted in further significant decrease in the angiogenic effects of ER-MC cells. Thus, ARTN stimulates de novo tumor angiogenesis mediated in part by VEGF-A. ARTN therefore co-ordinately regulates multiple aspects of tumor growth and metastasis.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Angiogenesis Inhibitors / pharmacology
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Animals
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Antibodies, Monoclonal, Humanized / pharmacology
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Antigens, CD / genetics
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Antigens, CD / metabolism
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Bevacizumab
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Breast Neoplasms / blood supply*
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Breast Neoplasms / genetics*
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Breast Neoplasms / metabolism
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Breast Neoplasms / pathology
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Cell Movement / drug effects
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Cell Proliferation / drug effects
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Endothelial Cells / drug effects
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Endothelial Cells / metabolism
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Endothelial Cells / pathology
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Endothelium, Vascular / drug effects
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Endothelium, Vascular / metabolism
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Endothelium, Vascular / pathology
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Female
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Gene Expression Regulation, Neoplastic / drug effects*
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Humans
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Mice
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Mice, Nude
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Neovascularization, Pathologic
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Nerve Tissue Proteins / antagonists & inhibitors
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Nerve Tissue Proteins / genetics*
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Nerve Tissue Proteins / metabolism
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism
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Proto-Oncogene Proteins c-akt / genetics
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Proto-Oncogene Proteins c-akt / metabolism
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RNA, Small Interfering / genetics
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Signal Transduction / drug effects*
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Twist-Related Protein 1 / genetics
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Twist-Related Protein 1 / metabolism
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Vascular Endothelial Growth Factor A / antagonists & inhibitors
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Vascular Endothelial Growth Factor A / genetics
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Vascular Endothelial Growth Factor A / metabolism
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Xenograft Model Antitumor Assays
Substances
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ARTN protein, human
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Angiogenesis Inhibitors
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Antibodies, Monoclonal, Humanized
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Antigens, CD
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Nerve Tissue Proteins
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Nuclear Proteins
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RNA, Small Interfering
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TWIST1 protein, human
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Twist-Related Protein 1
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VEGFA protein, human
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Vascular Endothelial Growth Factor A
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Bevacizumab
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Proto-Oncogene Proteins c-akt
Grants and funding
This work was funded by the Breast Cancer Research Trust, New Zealand, The Dick Roberts Trust, New Zealand, Cancer Science Institute, Singapore, The National Key Scientific Programme of China (2012CB934002 and 2010CB912804), Chinese Academy of Sciences (XDA01040410), National Natural Science Foundation of China (30971492, and 81101597), and the Chinese Academy of Sciences Visiting Professorship for Senior International Scientists (2010T2S03). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.