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Updated: Jun 9, 2026

Strategic Endothelial Cell Tube Formation Assay: Comparing Extracellular Matrix and Growth Factor Reduced Extracellular Matrix
08:46

Strategic Endothelial Cell Tube Formation Assay: Comparing Extracellular Matrix and Growth Factor Reduced Extracellular Matrix

Published on: August 14, 2016

Involvement of c-Jun N-terminal kinase and extracellular signal-regulated kinase 1/2 in EGF-induced angiogenesis

Kaikai Shen1, Yuchen Sheng, Lili Ji

  • 1Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201210, Peoples Republic of China.

Cell Biology International
|August 26, 2010
PubMed

Insights

Epidermal growth factor (EGF) triggers angiogenesis in human umbilical vein endothelial cells (HUVECs) primarily through the JNK/SAPK and ERK1/2 pathways, influencing both cell proliferation and tube formation.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Physiology

Background:

  • Angiogenesis is essential for forming new blood vessels from existing ones.
  • Mitogen-activated protein kinases (MAPKs) are implicated in cellular signaling pathways.
  • Epidermal growth factor (EGF) is a known stimulant of endothelial cell activity.

Purpose of the Study:

  • To investigate the role of MAPKs in EGF-induced angiogenesis in human umbilical vein endothelial cells (HUVECs).
  • To determine which specific MAPK pathways are critical for EGF-mediated endothelial cell proliferation and tube formation.

Main Methods:

  • HUVECs were treated with varying concentrations of EGF.
  • Western blot analysis was used to detect MAPK phosphorylation.
  • Specific MAPK inhibitors (SP600125, PD098059, SB203580) and siRNA were employed to block pathway activity.
  • Tube formation and cell proliferation assays were conducted.

Main Results:

  • EGF stimulated HUVEC proliferation in a dose-dependent manner, with 10 ng/ml being optimal.
  • EGF induced phosphorylation of ERK1/2, p38, and JNK/SAPK.
  • Inhibition of JNK/SAPK (using SP600125 or siRNA) significantly blocked EGF-induced tube formation and proliferation.
  • Inhibition of ERK1/2 partially blocked tube formation, while p38 inhibition had no effect.
  • SP600125 and JNK/SAPK siRNA showed no cytotoxicity at effective concentrations.

Conclusions:

  • JNK/SAPK signaling is crucial for EGF-induced HUVEC proliferation and angiogenesis.
  • ERK1/2 signaling also plays a role in EGF-mediated tube formation.
  • MAPK pathways, particularly JNK/SAPK, are key regulators of EGF-driven angiogenesis.

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