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Related Experiment Videos

Tissue transglutaminase mRNA expression in apoptotic cell death.

E B Volokhina1, R Hulshof, C Haanen

  • 1Department of Clinical Chemistry, Medisch Spectrum Twente, Hospital Group, P.O. Box 50000, 7500 KA Enschede, The Netherlands.

Apoptosis : an International Journal on Programmed Cell Death
|January 24, 2004
PubMed
Summary

Tissue transglutaminase messenger RNA (t.TG-mRNA) expression significantly increases with apoptosis. This rise in t.TG-mRNA levels can serve as a reliable marker for detecting and quantifying recent apoptotic cell death.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Tissue transglutaminase (t.TG) is an enzyme crucial for protein cross-linking and maintaining cellular integrity.
  • During apoptosis, t.TG is activated and plays a role in the formation of apoptotic bodies.
  • Understanding t.TG's role in apoptosis is key to developing new diagnostic markers.

Purpose of the Study:

  • To investigate the expression levels of t.TG-mRNA during apoptosis.
  • To determine if t.TG-mRNA expression can be used as a quantitative marker for recent apoptosis.
  • To assess the reliability and specificity of t.TG-mRNA as an apoptosis biomarker.

Main Methods:

  • Real-time RT-PCR (TaqMan based) was employed to measure t.TG-mRNA expression.
  • Experiments were conducted on cultured cells (MCF-7, human endothelial cells) and peripheral blood mononuclear cells (PBMCs).

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  • Apoptosis was induced in vitro, and t.TG-mRNA levels were measured before and after induction.

    • Main Results:

    • TaqMan RT-PCR for t.TG-mRNA was found to be reliable, reproducible, and specific for apoptotic cell death.
    • t.TG-mRNA expression increased significantly (up to 20-fold) in response to apoptosis induction in a dose- and time-dependent manner.
    • Elevated t.TG-mRNA expression was observed in PBMCs from myelodysplasia patients, correlating with in vivo apoptosis.

    Conclusions:

    • t.TG-mRNA expression is a significant indicator of apoptosis induction.
    • The dose and time-dependent nature of t.TG-mRNA changes supports its utility.
    • t.TG-mRNA serves as a valuable trace marker for the detection and quantification of apoptosis.