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Blood, Vol. 94 No. 3 (August 1), 1999:
pp. 959-967
Homocysteine-Induced Endoplasmic Reticulum Stress and Growth Arrest
Leads to Specific Changes in Gene Expression in Human Vascular
Endothelial Cells
P. Andrew Outinen,
Sudesh K. Sood,
Sabine I. Pfeifer,
Sushmita Pamidi,
Thomas J. Podor,
Jun Li,
Jeffrey I. Weitz, and
Richard C. Austin
From the Department of Pathology and Molecular Medicine, McMaster
University and the Hamilton Civic Hospitals Research Centre, Hamilton,
Ontario, Canada.
Alterations in the cellular redox potential by homocysteine promote
endothelial cell (EC) dysfunction, an early event in the progression of
atherothrombotic disease. In this study, we demonstrate that
homocysteine causes endoplasmic reticulum (ER) stress and growth arrest
in human umbilical vein endothelial cells (HUVEC). To determine if
these effects reflect specific changes in gene expression, cDNA
microarrays were screened using radiolabeled cDNA probes generated from
mRNA derived from HUVEC, cultured in the absence or presence of
homocysteine. Good correlation was observed between expression profiles
determined by this method and by Northern blotting. Consistent with its
adverse effects on the ER, homocysteine alters the expression of genes
sensitive to ER stress (ie, GADD45, GADD153, ATF-4, YY1). Several other genes observed to be differentially expressed by homocysteine are known
to mediate cell growth and differentiation (ie, GADD45, GADD153, Id-1,
cyclin D1, FRA-2), a finding that supports the observation that
homocysteine causes a dose-dependent decrease in DNA synthesis in
HUVEC. Additional gene profiles also show that homocysteine decreases
cellular antioxidant potential (glutathione peroxidase, NKEF-B PAG,
superoxide dismutase, clusterin), which could potentially enhance the
cytotoxic effects of agents or conditions known to cause oxidative
damage. These results successfully demonstrate the use of cDNA
microarrays in identifying homocysteine-respondent genes and indicate
that homocysteine-induced ER stress and growth arrest reflect specific
changes in gene expression in human vascular EC.
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C. H. He, P. Gong, B. Hu, D. Stewart, M. E. Choi, A. M. K. Choi, and J. Alam
Identification of Activating Transcription Factor 4 (ATF4) as an Nrf2-interacting Protein. IMPLICATION FOR HEME OXYGENASE-1 GENE REGULATION
J. Biol. Chem.,
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[Abstract]
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[PDF]
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N. Weiss, Y.-Y. Zhang, S. Heydrick, C. Bierl, and J. Loscalzo
Overexpression of cellular glutathione peroxidase rescues homocyst(e)ine-induced endothelial dysfunction
PNAS,
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[Abstract]
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[PDF]
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