15(S)-hydroxyeicosatetraenoic acid–induced angiogenesis requires Src-mediated Egr-1–dependent rapid induction of FGF-2 expression
Authors: Nikhlesh K Singh
Gadiparthi N Rao
Qiuhua Zhang
Jixiao Niu
Jaganathan Subramani
Saradasri Karri
Dong Wang
Dong Van Quyen
Venkatesh Kundumani-Sridharan
Journal: Blood (2010)
Discipline: CLINICAL MEDICINE
Keywords: 15(S)-HETE; Egr-1; angiogenesis; HDMVECs; Src
Affiliations: Department of Physiology, University of Tennessee Health Science Center, 894 Union Ave, Memphis, TN 38163, USA.

Abstract:  To understand the mechanisms underlying 15(S)-hydroxyeicosatetraenoic acid [15(S)-HETE]-induced angiogenesis, we studied the role of Egr-1. 15(S)-HETE induced Egr-1 expression in a time-dependent manner in human dermal microvascular endothelial cells (HDMVECs). Blockade of Egr-1 via forced expression of its dominant-negative mutant attenuated 15(S)-HETE-induced HDMVEC migration and tube formation as well as Matrigel plug angiogenesis. 15(S)-HETE-induced Egr-1 expression requires Src activation. In addition, adenovirus-mediated expression of dominant-negative mutant of Src blocked 15(S)-HETE's effects on migration and tube formation of HDMVECs and Matrigel plug angiogenesis. 15(S)-HETE induced fibroblast growth factor-2 (FGF-2) expression rapidly via Src-mediated production of Egr-1. Cloning and mutational analysis of FGF-2 promoter revealed that Egr-1 binding site proximal to transcription start site is required for 15(S)-HETE-induced FGF-2 expression. Neutralizing antibody-mediated suppression of FGF-2 function also attenuated the effects of 15(S)-HETE on HDMVEC migration and tube formation as well as Matrigel plug angiogenesis. Furthermore, in contrast to wild-type mice, 12/15-LOX(-/-) mice exhibited decreased Matrigel plug angiogenesis in response to AA, which was rescued by 15(S)-HETE. On the basis of these observations, we conclude that 15(S)-HETE-induced angiogenesis requires Src-mediated Egr-1-dependent rapid induction of FGF-2. These findings may suggest that 15(S)-HETE could be a potential endogenous regulator of pathologic angiogenesis associated with atherosclerosis and restenosis. 

Retracted: No

PubPeer comments:
 Part of Fig panel 1D and 2D look very similar: ![file] Fig 1. Egr-1 mediates 15(S)-HETEu2013induced HDMVEC migration and tube-like structure formation in vitro and Matrigel plug angiogenesis in vivo. Panel D: HDMVECs were transduced with Ad-GFP or Ad-dnEgr-1 at a moi of 40, quiesced, and subjected to 15(S)-HETEu2013induced tube-like structure formation Fig 2. Src mediates 15(S)-HETEu2013induced Egr-1 expression in HDMVECs, leading to their migration and tube-like structure formation. Panel D: HDMVECs that were transduced with Ad-GFP or Ad-dnSrc at a moi of 40 and quiesced were subjected to 15(S)-HETE (0.1u03bcM)u2013induced tube-like structure formation , 
 Dear Hyphoderma Heterocystidiatum: Thank you for posting a comment on our paper pointing similarity of one image between Figure 1D and Figure 2D. We performed the experiment to study the role of Egr1 and Src on 15(S)-HETE-induced EC tube formation in the same experiment but the data was split between Figure 1D and Figure 2D for better presentation, and the image for Ad-GFP + 15(S)-HETE treatment was inserted inadvertently in both Figure 1D and Figure 2D. Since this is one of the several images of Ad-GFP + 15(S)-HETE treatment groups that were used for comparing the effects of dn-Egr1 + 15(S)-HETE and dn-Src + 15(S)-HETE treatments from the same experiment and the bar graphs show quantitative data of several replicates with statistical analysis, it does not affect the results or the conclusions of the paper. Thank you Venkatesh Kundumani-Sridharan ,