Increased expression of extracellular proteins as a hallmark of human endothelial cell in vitro senescence

Exp Gerontol. 2006 May;41(5):474-81. doi: 10.1016/j.exger.2006.03.001. Epub 2006 Apr 19.

Abstract

A convenient way to study processes of aging in distinct human tissues consists of a molecular analysis of cells from the tissue in question, that were explanted and grown in vitro until they reach senescence. Using human umbilical vein endothelial cells (HUVEC), we have established an in vitro senescence model for human endothelial cells. A major hallmark of HUVEC in vitro senescence is the increased frequency of apoptotic cell death, which occurs as a determining feature of HUVEC senescence. Senescent endothelial cells are also found in vivo in atherosclerotic lesions, suggesting that the presence of such cells may contribute to the development of vascular pathology. To elucidate mechanisms underlying endothelial cell senescence and age-associated apoptosis, gene expression analyses were carried out. In these experiments, we observed the up-regulation of genes coding for extracellular proteins in senescent HUVEC. In particular, a significant upregulation of interleukin-8, VEGI, and the IGF-binding proteins 3 and 5 was observed. Upregulation of these genes was confirmed by both RT-PCR and Western blot. In the case of interleukin-8, a roughly 50-fold upregulation of the protein was also found in cellular supernatants. The extracellular proteins encoded by these genes are well known for their ability to modulate the apoptotic response of human cells, and in the case of interleukin-8, clear links to the establishment of atherosclerotic lesions have been defined. The results described here support a new model, where changes in the secretome of human endothelial cells contribute to vascular aging and vascular pathology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis
  • Blotting, Western / methods
  • Cells, Cultured
  • Cellular Senescence / physiology*
  • Endothelial Cells / metabolism
  • Endothelial Cells / physiology*
  • Endothelium, Vascular / cytology*
  • Endothelium, Vascular / metabolism
  • Humans
  • Insulin-Like Growth Factor Binding Proteins / biosynthesis
  • Insulin-Like Growth Factor Binding Proteins / genetics
  • Interleukin-8 / biosynthesis
  • Interleukin-8 / genetics
  • Protein Biosynthesis / physiology*
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Tumor Necrosis Factor Ligand Superfamily Member 15 / biosynthesis
  • Tumor Necrosis Factor Ligand Superfamily Member 15 / genetics
  • Umbilical Veins / cytology
  • Umbilical Veins / metabolism
  • Up-Regulation / physiology

Substances

  • Insulin-Like Growth Factor Binding Proteins
  • Interleukin-8
  • TNFSF15 protein, human
  • Tumor Necrosis Factor Ligand Superfamily Member 15