Gene Expression Profile of Zinc-Deficient, Homocysteine-Treated Endothelial Cells

  • Kwun, In-Sook ;
  • Beattie, John H.
  • Published : 2003.12.01


In the post-genome period, the technique for identifying gene expression has been progressed to high throughput screening. In the field of molecular nutrition, the use of screening techniques to clarify molecular function of specific nutrients would be very advantageous. In this study, we have evaluated Zn-regulated gene expression in Zn-deficient, homocystein-treated EA.hy926 cells, using cDNA microarray, which can be used to screen the expression of many genes simultaneously. The information obtained can be used for preliminary assessment of molecular and signaling events modulated by Zn under pro-atherogenic conditions. EA.hy926 cells derived from human umbilical vein endothelial cells were cultured in Zn-adequate (control, 15 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) Dulbecco's MEM media under high homocysteine level (100 $\mu$M) for 3 days of post-confluency. Cells were harvested and RNA was extracted. Total RNA was reverse-transcribed and the synthesized cDNA was labeled with Cy3 or Cy5. Fluorescent labeled cDNA probe was applied to microarray slides for hybridization, and the slide was then scanned using a fluorescence scanner. The expression of seven genes was found to be significantly decreased, and one significantly increased, in response to treatment of EA.hy926 cells with Zn-deficient medium, compared with Zn-supplemented medium. The upregulated genes were oncogenes and tumor suppressor genes, cell cycle-related genes and transporter genes. The down-regulated gene was RelB, a component of the NF-kappaB complex of transcription factors. The results of this study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, namely Zn. Furthur study, using tailored-cDNA array and vascular endothelial cell lines, would be beneficial to clarify the molecular function of Zn in atherosclerosis, more in detail.


Zn-deficiency;endothelial cells;EA.hy926 cell;homocysteine;cDNA microarray;atherosclerosis


  1. J Nutr v.130 Human zinc deficiency Hambidge M
  2. Ann Rev Bioche v.61 Zinc proteins: enzymes, storage proteins, transcription factors, and replicationl proteins Coleman JE
  3. In Handbook of nutritionall essential mineral elements Chesters JK;O'Dell BL(ed.);Sunde RA(ed.)
  4. J Cell Physiol v.155 Two zinc-dependent steps during G1 to S phase transition Chesters JK;Petrie L;Lipson KE
  5. J Endocrinol v.178 Experiments using microarray technology: limitations and standard operating procedures Forster T;Roy D;Ghazal P
  6. Pharmacogenomics v.4 The anlysis of microarray data Hariharan R
  7. In The world Health Report 2002 Quantifying Selected Major Risks to Health WHO
  8. Bibliotheca Nutrito et Dieta v.54 Trace elements and cardiovascular disease Strain JJ
  9. Cancer Metastasis Rev v.21 Zinc in cancer prevention Prasad AS;Kucuk O
  10. Nutrition v.15 Zinc nutrition and apoptosis of vascular endothelial cells; implications in atherosclerosis Hennig B;Meerarani P;Ramadass P;Toborek M;Malecki A;Slim R;McClain CJ
  11. Brit J Nutr v.91 no.2 In zinc deficiency a risk factor for atherosclerosis? Beattie JH;Kwun IS
  12. Am J Clin Nutr v.71 Zinc protects against apoptosis of endothelial cells induced by linoleic acid and tumor necrosis factor alpha Meerarani P;Ramadass P;Toborek M;Bauer HC;Bauer H;Henning B
  13. Atherosclerosis. Gene Expression, Cell Interactions and Oxidation The endothelial cell in atherosclerosis Bannon P;James N;Jessup W;Dean RT(ed.);Kelly DT(ed.)
  14. J Nutr Health Aging v.5 Homocysteine, folic acid, B vitamins and cardiovascular risk Blacher J;Safar ME
  15. Atherosclerosis v.168 Homocysteine strongly enhances metal-catalyzed LDL oxidation in the presence of cystine and cysteine Pfanzagl B;Tribl F;Koller E;Moslinger T
  16. J Inorg Biochem v.95 Copperhomocysteine complexes and potential physiological actions Apostolova MD;Bontchev PR;Ivanova BB;Russell WR;Mehandjiev DR;Beattie JH;Nachev CK
  17. Proc Natl Acad Sci USA v.90 Ligand-specific activation of HER4/p180erbB4, a fourth member of the epidermal growth factor receptor family Plowman GD;Culouscou JM;Whitney GS;Green JM;Carlton GW;Foy L;Neubauer MG;Shoyab M
  18. Am J Physiol v.277 Neuregulin activation of ErbB receptors in vascular endothelium leads to angiogenesis Russell KS;Stern DF;Polverini PJ;Bender JR
  19. Antioxdants and Redox Signaling v.3 Zinc binding and redox control of p53 structure and function Hainaut P;Mann K
  20. Nature v.389 A model for p53-induced apoptosis Polyak K;Xia Y;Zweier JL;Kinzler KW;Vogelstein B
  21. Apoptosis v.3 Biological significance and molecular mechanisms of p53-induced apoptosis Bedi A;Mookerjee B
  22. proc Natl Acad Sci USA v.89 Cloning and antisense oligodeoxy-nucleotide inhibition of a human homolog of cdc2 required in hematopoiesis Lapidot-Lifson Y;Patinkin D;Prody CA;Ehrlich G;Seidman S:Ben Aziz R;Benseler F;Eckstein F;Zakut H;Soreq H
  23. Mol Cell Biol v.22 TRRAP-dependent and TRRAP-independent transcriptional activation by Myc family oncoproteins Nikiforov MA;Chandriani S;Park J;Kotenko I;Matheos D;Johnsso A;McMahon SB;Cole MD
  24. Immunology Today v.19 Signal transduction through NF-kappa May MJ;Ghosh S
  25. Ann N Y Acad Sci v.987 The role of relB in regulating the adaptive immune response Zanetti M;Castiglioni P;Schoenberger S;Gerloni M
  26. Bioch Biophy Res Comm v.259 Pyrithione, a zinc ionophore, inhibits NF-kappaB activation Kim CH;Kim JH;Moon SJ;Chung KC;Hsu CY;Seo JT;Ahn YS