University of Natural Resources and Life Sciences, Vienna (BOKU) - Research portal
Selected Publication:
Grillari, J; Hohenwarter, O; Grabherr, RM; Katinger, H; .
(2000):
Subtractive hybridization of mRNA from early passage and senescent endothelial cells.
Exp Gerontol. 2000; 35(2):187-197
- Abstract:
- Regulation of cellular processes that eventually lead to a state of growth arrest is an important manifestation of in vitro cellular senescence caused and accompanied by variations of the gene expression pattern. Whereas these changes at the mRNA level have been studied mainly in fibroblast cultures, we concentrated on endothelial cells that represent an accepted model for Vascular systems and may be involved in the pathogenesis of diseases related to aging. To isolate differentially expressed genes, we created a subtractive cDNA library using mRNA from senescent (35 passages) and young (five passages) human umbilical vein endothelial cells (HUVECs). Candidate clones were isolated from the cDNA library, differential expression was confirmed by Northern blot analyses and sequences were compared with a genbank data base. Because many mRNAs were below the detection limit of Northern blot analysis, we were forced to establish a more sensitive PCR based method (ATAC-PCR) to quantify and confirm altered levels of gene expression. Several mRNAs were found to be upregulated in senescent HUVECs including two components of the extracellular matrix (ECM): plasminogen activator inhibitor and fibronectin. Elevated expression of both has already been described in senescent cells. The mRNAs of TGF-beta-inducible gene H3 (beta-IG-H3; ECM protein), insulin-like growth factor binding protein (IGFBP-3), p53-inducible gene (PIG3) a protein involved in vesicular transport (SEC13R) and ribosomal protein L28 have likewise been shown to be preferentially expressed in senescent cells. Because studies support the involvement of ECM components, TGF-beta and p53 in tumor suppressing mechanisms, our data supports the hypothesis that cellular senescence and upregulation of ECM proteins may be associated with tumor preventive functions. (C) 2000 Elsevier Science Inc. All rights reserved.
- Authors BOKU Wien:
- Grabherr Reingard
- Grillari Johannes
- Katinger Hermann
- Find related publications in this database (using NML MeSH Indexing)
- Cell Aging/physiology*;Cells, Cultured;Endothelium, Vascular/cytology;Endothelium, Vascular/metabolism;Endothelium, Vascular/physiology*;Humans;Nucleic Acid Hybridization;Polymerase Chain Reaction/methods;RNA, Messenger/metabolism*;Subtraction Technique;
- Find related publications in this database (Keywords)
- human umbilical vein endothelial cells (HUVEC)
- replicative senescence
- subtractive hybridization
- ATAC-PCR
- TGF-beta inducible gene human 3 (beta IG-H3)
- SEC13R
- p53 inducible gene 3 (PIG3)
- fibronectin
- L6
- TAX REB 107
- IGFBP-3 PAI-1
- Staf50
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