Gamabufotalin, a major derivative of bufadienolide, inhibits VEGF-induced angiogenesis by suppressing VEGFR-2 signaling pathway

Ning Tang; Lei Shi; Zhenlong Yu; Peipei Dong; Chao Wang; Xiaokui Huo; Baojing Zhang; Shanshan Huang; Sa Deng; Kexin Liu; Tonghui Ma; Xiaobo Wang; Lijun Wu; Xiao-Chi Ma

doi:10.18632/oncotarget.6514

Gamabufotalin, a major derivative of bufadienolide, inhibits VEGF-induced angiogenesis by suppressing VEGFR-2 signaling pathway

Oncotarget. 2016 Jan 19;7(3):3533-47. doi: 10.18632/oncotarget.6514.

Authors

Ning Tang¹, Lei Shi², Zhenlong Yu^{1

3}, Peipei Dong¹, Chao Wang¹, Xiaokui Huo¹, Baojing Zhang¹, Shanshan Huang¹, Sa Deng¹, Kexin Liu¹, Tonghui Ma⁴, Xiaobo Wang³, Lijun Wu³, Xiao-Chi Ma^{1

3}

Affiliations

¹ College of Pharmacy, Academy of Integrative Medicine, Key Laboratory of Pharmacokinetic and Drug Transport of Liaoning, Dalian Medical University, Dalian, China.
² Institute of Cancer Stem Cell, Dalian Medical University, Dalian, China.
³ Department of Pharmacy and Traditional Chinese medicine, Chinese People's Liberation Army 210 Hospital, Dalian, China.
⁴ College of Basic Medical Science, Dalian Medical University, Dalian, China.

Abstract

Gamabufotalin (CS-6), a main active compound isolated from Chinese medicine Chansu, has been shown to strongly inhibit cancer cell growth and inflammatory response. However, its effects on angiogenesis have not been known yet. Here, we sought to determine the biological effects of CS-6 on signaling mechanisms during angiogenesis. Our present results fully demonstrate that CS-6 could significantly inhibit VEGF triggered HUVECs proliferation, migration, invasion and tubulogenesis in vitro and blocked vascularization in Matrigel plugs impregnated in C57/BL6 mice as well as reduced vessel density in human lung tumor xenograft implanted in nude mice. Computer simulations revealed that CS-6 interacted with the ATP-binding sites of VEGFR-2 using molecular docking. Furthermore, western blot analysis indicated that CS-6 inhibited VEGF-induced phosphorylation of VEGFR-2 kinase and suppressed the activity of VEGFR-2-mediated signaling cascades. Therefore, our studies demonstrated that CS-6 inhibited angiogenesis by inhibiting the activation of VEGFR-2 signaling pathways and CS-6 could be a potential candidate in angiogenesis-related disease therapy.

Keywords: angiogenesis; aortic ring; gamabufotalin; vascular endothelial growth factor (VEGF); vascular endothelial growth factor receptor 2 (VEGFR-2).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Angiogenesis Inhibitors / chemistry
Angiogenesis Inhibitors / pharmacology*
Animals
Aorta / cytology
Aorta / metabolism
Apoptosis
Blotting, Western
Bufanolides / chemistry
Bufanolides / pharmacology*
Cell Movement
Cell Proliferation
Cells, Cultured
Endothelium, Vascular / cytology
Endothelium, Vascular / drug effects*
Endothelium, Vascular / metabolism
Fluorescent Antibody Technique
Human Umbilical Vein Endothelial Cells / cytology
Human Umbilical Vein Endothelial Cells / drug effects*
Human Umbilical Vein Endothelial Cells / metabolism
Humans
Immunoprecipitation
Lung Neoplasms / blood supply
Lung Neoplasms / drug therapy*
Lung Neoplasms / metabolism
Lung Neoplasms / pathology
Mice
Mice, Inbred C57BL
Mice, Nude
Models, Molecular
Neovascularization, Physiologic / drug effects*
RNA, Small Interfering / genetics
Vascular Endothelial Growth Factor A / chemistry
Vascular Endothelial Growth Factor A / metabolism
Vascular Endothelial Growth Factor Receptor-2 / antagonists & inhibitors*
Vascular Endothelial Growth Factor Receptor-2 / genetics
Vascular Endothelial Growth Factor Receptor-2 / metabolism
Wound Healing

Substances

Angiogenesis Inhibitors
Bufanolides
RNA, Small Interfering
VEGFA protein, human
Vascular Endothelial Growth Factor A
gamabufotalin
Vascular Endothelial Growth Factor Receptor-2