Inhibition of endothelial cell proliferation, in vitro angiogenesis, and the down-regulation of cell adhesion-related genes by genistein. Combined with a cDNA microarray analysis.

Abstract

Antiangiogenesis is presently one of the powerful strategies for treating cancer, and endothelial cells play a pivotal role in the process of angiogenesis. Genistein, a tyrosine kinase inhibitor, a major isoflavone plentiful in soybeans, is known to inhibit both tumor growth and angiogenesis. However, the precise molecular mechanism(s) by which genistein affects endothelial cells has yet to be elucidated. In the present study, a cDNA microarray was performed to investigate the targeted genes of human umbilical vein endothelial cells (HUVECs) affected by 10 microM genistein. As a result, a total of 256 genes showed an altered expression of more than twofold. Among them were the genes related to cell proliferation, adhesion, transcription, translation, metabolism, cytoskeleton, apoptosis, kinases, and functionally unknown. The down-regulation of mRNA or the protein expression of cell adhesion-related genes, including VE-cadherin, gap junction protein alpha 1 (connexin 43), integrin alpha V, and multimerin, were confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) or by immunofluorescence staining. The impaired cell-cell adhesion by genistein was also observed by electron microscopy. In addition, the antiangiogenesis role of genistein was confirmed on Matrigel using inverted microscopy and electron microscopy. In conclusion, genistein affects endothelial cells as a negative mediator of proliferation and angiogenesis in vitro, partially by down-regulating cell adhesion-related genes and impairing cell adhesions.