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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2003;23:231.)
© 2003 American Heart Association, Inc.

Vascular Biology

HEX Acts as a Negative Regulator of Angiogenesis by Modulating the Expression of Angiogenesis-Related Gene in Endothelial Cells In Vitro

Tomowaki Nakagawa; Mayumi Abe; Tohru Yamazaki; Hiroki Miyashita; Hitoshi Niwa; Shoichi Kokubun; Yasufumi Sato

From the Department of Vascular Biology (T.N., M.A., T.Y., H.M., Y.S.), Institute of Development, Aging and Cancer, Tohoku University, Sendai; the Department of Orthopedic Surgery (T.N., S.K.), Tohoku University Graduate School of Medicine, Sendai; and the Laboratory for Cell Pluripotency Studies (H.N.), RIKEN Center for Developmental Biology, Kobe, Japan.

Correspondence to Yasufumi Sato, Department of Vascular Biology, Institute of Development, Aging and Cancer, Tohoku University, 4-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan. E-mail y-sato{at}idac.tohoku.ac.jp

Objective— The hematopoietically expressed homeobox (HEX) is transiently expressed in endothelial cells (ECs) during vascular formation in embryo. Here, we investigated whether HEX played any role in angiogenesis-related properties of ECs in vitro.

Methods and Results— We transiently overexpressed HEX in human umbilical vein ECs (HUVECs). To our surprise, HEX completely abrogated the response of HUVECs to vascular endothelial growth factor (VEGF) with regard to proliferation, migration, and invasion and abolished network formation by HUVECs on Matrigel. cDNA microarray analysis and quantitative real-time reverse transcription–polymerase chain reaction combined with Western blotting revealed that HEX significantly repressed the expression of VEGF receptor-1, VEGF receptor-2, neuropilin-1, tyrosine kinase with Ig and EGF homology domains (TIE)-1, TIE-2, and the integrin v subunit, whereas it augmented the expression of endoglin in HUVECs. We established murine embryonic stem cells that were stably transfected with HEX sense cDNA or antisense cDNA, and we examined the in vitro differentiation to ECs. Although the expression of VEGF receptor-2 was decreased in sense transfectants, the number of cells expressing VE-cadherin, a specific marker of ECs, was not altered.

Conclusions— Our present results suggest that HEX may not affect the differentiation of ECs but acts as a negative regulator of angiogenesis.

Key Words: angiogenesis • HEX • transcription factor • endothelial cell

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