Abstract
Background
Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of
growth factors that have been implicated in skin patho-physiology. Although endothelial
nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) appear
to be involved in mitogenesis and chemotaxis in epidermal keratinocytes, the activation
of eNOS and VEGF production induced by HB-EGF and its signaling mechanism remains
undefined.
Objective
We examined possible signal transduction pathways by which HB-EGF leads to eNOS activation
and VEGF production in human epidermal keratinocyte cell line (HaCaT cells).
Methods
The phosphorylation of epidermal growth factor receptor (EGFR), mitogen-activated
protein kinase (MAPK; p42/p44 MAPK), Akt and eNOS were examined by Western blotting
analysis. VEGF production was determined by enzyme-linked immunosorbent assay. Various
inhibitors were utilized to investigate the signaling mechanisms of eNOS activation
and VEGF production.
Results
HB-EGF-induced phosphorylation of EGFR with maximum phosphorylation at 1 h. HB-EGF-induced phosphorylation of p42/p44 MAPK in a few minutes. It activated Akt
with maximum phosphorylation at 1 h and eNOS with maximum phosphorylation at 3 h. The HB-EGF-induced eNOS activation was significantly blocked by the p42/p44 MAPK
inhibitor U0126 and the phosphatidylinositol 3-kinase (P13K) inhibitor LY294002. HB-EGF
increased VEGF production. The HB-EGF-induced VEGF production was blocked by U0126
and LY294002. Finally, the HB-EGF-induced activation of Akt and eNOS was suppressed
by VEGF competitive antagonist, CBO-P11.
Conclusion
These results demonstrate that HB-EGF-induced eNOS activation depends on p42/p44 MAPK,
PI3K/Akt pathways and endogenous VEGF in HaCaT cells.
Keywords
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Article info
Publication history
Published online: July 12, 2010
Accepted:
June 2,
2009
Received in revised form:
May 21,
2009
Received:
March 25,
2009
Identification
Copyright
© 2009 Japanese Society for Investigative Dermatology. Published by Elsevier Inc. All rights reserved.
