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Thromb Haemost 2002; 88(06): 1047-1053
DOI: 10.1055/s-0037-1613353
Involvement of Thrombin Receptors in the Subject-dependent Variability in Ca2+ Signal Generation
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Flow Conditions Modulate Homocysteine Induced Changes in the Expression of Endothelial Cell Genes Associated with Cell-Cell Interaction and Cytoskeletal Rearrangement

Rima Dardik
1   Institute of Thrombosis and Hemostasis, Sheba Medical Center, Tel Hashomer
,
Naphtali Savion
2   Goldschleger Eye Research Institute, Sackler Medical School, Tel Aviv University
,
Nurit Gal
3   Functional Genomics Unit, Molecular Hemato-Oncology & Institute of Respiratory Medicine, Sheba Medical Center, Tel Hashomer
,
David Varon
4   Coagulation Unit, Department of Hematology, Hadassah, Hebrew University Medical Center, Jerusalem, Israel
› Author Affiliations
Further Information

Publication History

Received 18 February 2002

Accepted after resubmission 28 August 2002

Publication Date:
09 December 2017 (online)

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Summary

In the present study we analyzed changes in the pattern of gene expression in endothelial cells (HUVEC) exposed to elevated homocysteine levels under flow conditions. RNA samples of untreated and homocysteine treated EC were analyzed using a human cDNA array containing 588 cDNAs relevant to the cardiovascular system. Exposure to homocysteine under flow resulted in altered expression of 8 genes, the alterations of 3 of which were further confirmed by RT-PCR analysis: upregulation of MCP-1 and of profilin-I, and downregulation of alpha-catenin. The increased expression of profilin-I and the decreased expression of alpha-catenin were also confirmed at the protein level by Western blot analysis. Furthermore, immunostaining for alpha-catenin demonstrated complete loss of the protein from intercellular junctions following exposure to homocysteine under flow. All these changes were not observed in cells exposed to homocysteine under static conditions. We show that under flow conditions, homocysteine modulates the expression of genes/proteins involved in actin cytoskeleton reorganization, as well as in the formation and/or maintenance of cellcell junction, thus contributing to the morphological changes (e.g. cell retraction) reflecting endothelial cell injury.

Keywords

Homocysteine - endothelial cells - cell-cell junctions - alpha-catenin - profilin
 

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