Variability in E-selectin expression, mRNA levels and sE-selectin release between endothelial cell lines and primary endothelial cells

Cell Biol Int. 2000;24(2):91-9. doi: 10.1006/cbir.1999.0455.

Abstract

Endothelial cell lines express markers and are assumed to exhibit other endothelial cell responses. We investigated E-selectin expression from human umbilical vein endothelial cells, the spontaneously transformed ECV304 line and the hybrid line EA.hy926 by flow cytometry and immunofluorescence, mRNA and soluble E-selectin release. In cells exposed to tumour necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta), median (range) percentage of E-selectin-positive HUVECs increased from 1.6(0.9-6. 2)% to 91.4(83.0-96.1)%, (P=0.001) using flow cytometry. In contrast, E-selectin expression by ECV304 and EA.hy926 cell lines was 100-fold lower. E-selectin mRNA was detectable after 2 h, maximal at 6 h in HUVECs and undetectable in EA.hy926 and ECV304 cell lines after exposure to TNF-alpha/IL-1beta. sE-selectin accumulation increased (P=0.004) in HUVECs only. Neutrophil adherence to ECV304 and EA.hy926 cells was poor compared to HUVECs (P=0.004). The cell lines ECV304 and EA.hy926 do not exhibit normal endothelium expression of E-selectin, and may not be appropriate for studies of adhesion.

MeSH terms

  • Cell Adhesion
  • Cell Line
  • Cells, Cultured
  • E-Selectin / biosynthesis*
  • Endothelium, Vascular / metabolism*
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Humans
  • Interleukin-1 / pharmacology
  • Neutrophils / metabolism
  • RNA, Messenger / metabolism
  • Time Factors
  • Tumor Necrosis Factor-alpha / pharmacology
  • Umbilical Cord / metabolism
  • von Willebrand Factor / biosynthesis

Substances

  • E-Selectin
  • Interleukin-1
  • RNA, Messenger
  • Tumor Necrosis Factor-alpha
  • von Willebrand Factor