Research

Analysis of VEGF-responsive Genes Involved in the activation of endothelial cells

Kishore K Wary¹ , Geeta D Thakker² , Joseph O Humtsoe¹ and Jun Yang¹

¹  Center for Extracellular Matrix Biology, Institute of Biosciences and Technology, Texas A&M University System-Health Science Center, 2121 W. Holcombe Blvd., Houston, TX-77030, USA

²  Current address: Lexicon Genetics Inc., 8800 Technology Forest Place, The Woodlands, TX-77381

author email corresponding author email

Molecular Cancer 2003, 2:25doi:10.1186/1476-4598-2-25

Published:	9 July 2003

Abstract

Background

Identification of the genes and pathways associated with the activation of endothelial cells (ECs) could help uncover the role of ECs in wound healing, vascular permeability, blood brain barrier function, angiogenesis, diabetic retinopathy, atherosclerosis, psoriasis, and growth of solid tumors.

Design

Herein, we embedded ECs in 3D type I collagen gel, left unstimulated or stimulated with VEGF¹⁶⁵, and subjected to suppression subtractive hybridization followed by differential display (SSHDD). Gene fragments obtained from SSHDD were subjected to DNA sequence analysis. Database search with nucleotide sequence were performed using the BLAST algorithm and expression of candidate genes determined by northern blot analysis.

Results

A total of ~32 cDNA fragments, including known regulators of angiogenesis, and a set of genes that were not reported to be associated with activation of ECs and angiogenesis previously were identified. We confirmed the mRNA expression of KDR, α₂ integrin, Stanniocalcin, including a set of 11 candidate genes. Western immunoblotting results indicated that KDR, α₂ integrin, MMP-1, MMP-2, and VE-cadherin genes were indeed active genes.

Conclusion

We have identified a set of 11 VEGF-responsive endothelial cell candidate genes. Their expression in endothelial cell is confirmed by northern blot analyses. This preliminary report forms as a foundation for functional studies to be performed to reveal their roles in EC activation and pathophysiological events associated with the vasculature including tumor growth.