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Updated: Jun 2, 2026
HUVEC Tube-formation Assay to Evaluate the Impact of Natural Products on Angiogenesis
Published on: June 24, 2019
Cyclopeptide RA-V inhibits angiogenesis by down-regulating ERK1/2 phosphorylation in HUVEC and HMEC-1 endothelial
Grace G L Yue1, Jun-Ting Fan, Julia K M Lee
1Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
Background And Purpose:
Anti-angiogenic agents have recently become one of the major adjuvants for cancer therapy. A cyclopeptide, RA-V, has been shown to have anti-tumour activities. Its in vitro anti-angiogenic activities were evaluated in the present study, and the underlying mechanisms were also assessed.
Experimental Approach:
Two endothelial cell lines, human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC-1), were used. The effects of RA-V on the proliferation, cell cycle phase distribution, migration, tube formation and adhesion were assessed. Western blots and real-time PCR were employed to examine the protein and mRNA expression of relevant molecules.
Key Results:
RA-V inhibited HUVEC and HMEC-1 proliferation dose-dependently with IC(50) values of 1.42 and 4.0 nM respectively. RA-V inhibited migration and tube formation of endothelial cells as well as adhesion to extracellular matrix proteins. RA-V treatment down-regulated the protein and mRNA expression of matrix metalloproteinase-2. Regarding intracellular signal transduction, RA-V interfered with the activation of ERK1/2 in both cell lines. Furthermore, RA-V significantly decreased the phosphorylation of JNK in HUVEC whereas, in HMEC-1, p38 MAPK was decreased.
Conclusions And Implications:
RA-V exhibited anti-angiogenic activities in HUVEC and HMEC-1 cell lines with changes in function of these endothelial cells. The underlying mechanisms of action involved the ERK1/2 signalling pathway. However, RA-V may regulate different signalling pathways in different endothelial cells. These findings suggest that RA-V has the potential to be further developed as an anti-angiogenic agent.
Insights
The cyclopeptide RA-V demonstrates significant anti-angiogenic properties by inhibiting endothelial cell proliferation and migration. Its mechanisms involve the ERK1/2 signaling pathway, suggesting potential as a novel cancer therapy adjuvant.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Anti-angiogenic agents are crucial in cancer therapy.
- The cyclopeptide RA-V exhibits anti-tumor activities.
- In vitro anti-angiogenic potential of RA-V requires evaluation.
Purpose of the Study:
- To assess the in vitro anti-angiogenic activities of RA-V.
- To investigate the underlying molecular mechanisms of RA-V's action.
- To evaluate RA-V's effects on endothelial cell functions.
Main Methods:
- Utilized human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC-1).
- Assessed proliferation, cell cycle, migration, tube formation, and adhesion.
- Employed Western blots and real-time PCR for molecular analysis.
Main Results:
- RA-V inhibited endothelial cell proliferation (IC50: 1.42 nM for HUVEC, 4.0 nM for HMEC-1).
- RA-V reduced endothelial cell migration, tube formation, and adhesion.
- RA-V modulated matrix metalloproteinase-2 expression and ERK1/2 signaling.
Conclusions:
- RA-V exhibits potent anti-angiogenic activity in endothelial cells.
- Mechanisms involve the ERK1/2 pathway, with potential differential effects on other pathways.
- RA-V shows promise for development as an anti-angiogenic cancer therapeutic.
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