Establishment of a quantitative RT-pCR for detection of vascular cell adhesion molecule-1 transcripts in endothelial cells after stimulation with advanced glycation endproducts.

  • Authors:
    • T Kunt
    • T Forst
    • A Wilhelm
    • A Zschäbitz
    • A Göpfert
    • P Kann
    • A Pfuetzner
    • O Harzer
    • M Engelbach
    • J Beyer
    • E Stofft
  • View Affiliations

  • Published online on: October 1, 1998     https://doi.org/10.3892/ijmm.2.4.455
  • Pages: 455-515
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Abstract

Advanced glycation endproducts (AGE) are supposed to increase endothelial expression of adhesion molecules like vascular cell adhesion molecule-1 (VCAM-1) by inducing an intracellular stress with subsequent activation of nuclear transcription factor NF-kappa-B. Quantitative analysis of VCAM-1-transcription has not been demonstrated concerning this topic. Thus, the aim of this study was to establish quantitative reverse transcription polymerase chain reaction (RT-PCR) assays using a spacer gene in order to measure the amounts of specific mRNA for VCAM-1 in human umbilical vein endothelial cells (HUVEC) which were stimulated with AGE-albumin (AGE-BSA). A recombinant RNA-standard was synthesized and used as internal RT-PCR standard. The amount of VCAM-1-mRNA in unstimulated HUVEC was found to be 2.2 +/- 2.7 copies per cell. After stimulation with AGE-BSA, mRNA-levels were elevated to 38.9 +/- 10.9 copies per cell. Positive controls (stimulated with lipopolysaccharide) revealed mRNA-levels of 78.7 +/- 27.5 copies per cell. We conclude that quantitative RT-PCR using the spacer gene technique is a valid and reliable method for the measurement of small amounts of specific

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Oct 1998
Volume 2 Issue 4

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Kunt T, Forst T, Wilhelm A, Zschäbitz A, Göpfert A, Kann P, Pfuetzner A, Harzer O, Engelbach M, Beyer J, Beyer J, et al: Establishment of a quantitative RT-pCR for detection of vascular cell adhesion molecule-1 transcripts in endothelial cells after stimulation with advanced glycation endproducts.. Int J Mol Med 2: 455-515, 1998
APA
Kunt, T., Forst, T., Wilhelm, A., Zschäbitz, A., Göpfert, A., Kann, P. ... Stofft, E. (1998). Establishment of a quantitative RT-pCR for detection of vascular cell adhesion molecule-1 transcripts in endothelial cells after stimulation with advanced glycation endproducts.. International Journal of Molecular Medicine, 2, 455-515. https://doi.org/10.3892/ijmm.2.4.455
MLA
Kunt, T., Forst, T., Wilhelm, A., Zschäbitz, A., Göpfert, A., Kann, P., Pfuetzner, A., Harzer, O., Engelbach, M., Beyer, J., Stofft, E."Establishment of a quantitative RT-pCR for detection of vascular cell adhesion molecule-1 transcripts in endothelial cells after stimulation with advanced glycation endproducts.". International Journal of Molecular Medicine 2.4 (1998): 455-515.
Chicago
Kunt, T., Forst, T., Wilhelm, A., Zschäbitz, A., Göpfert, A., Kann, P., Pfuetzner, A., Harzer, O., Engelbach, M., Beyer, J., Stofft, E."Establishment of a quantitative RT-pCR for detection of vascular cell adhesion molecule-1 transcripts in endothelial cells after stimulation with advanced glycation endproducts.". International Journal of Molecular Medicine 2, no. 4 (1998): 455-515. https://doi.org/10.3892/ijmm.2.4.455