Hypoxia‐Induced Membrane‐Bound Apoptotic DNA Particles
Annals of the New York Academy of Sciences(2006)
摘要
Abstract: Fetal DNA is found in the plasma of pregnant women that appears to be stable for PCR amplification. Although the underlying mechanism giving rise to this DNA in plasma remains unclear, the source of these fragments may be from apoptotic bodies (Apo‐Bodies) created from dying cells. Trophoblast apoptosis is essential for normal placental development, given the enormous amount of proliferation, differentiation, and migration during pregnancy. Through flow cytometric analysis coupled with real‐time PCR, our lab has shown that aggregates of acridine orange (AO)‐stained material (apoptotic particles) are resistant to DNase treatment, disrupted by sodium dodecyl sulfate (SDS), and contain fetal DNA. Because the placenta continuously remodels in an hypoxic environment, our hypothesis is that fetal DNA in maternal plasma comes from hypoxia‐induced dying trophoblasts and that this DNA circulates predominately in the form of Apo‐Bodies. We have developed a model culture system for analysis of Apo‐Bodies derived from JEG‐3 cells, an extravillous trophoblastic cell line, undergoing various methods of cell death: hypoxia‐induced, etoposide‐induced, and heat stress (necrosis like)–induced cell death. Under conditions of similar propidium iodide (PI) uptake, suggesting comparable levels of death, both hypoxia‐ and etoposide‐induced Apo‐Bodies increase in concentration over time, whereas heat‐induced levels of particles remain fairly constant, indicating that production of DNA‐associated Apo‐Bodies is a continuous process. Hypoxia, which is likely to be responsible for trophoblast cell death in vivo, produced membrane‐bound Apo‐Bodies containing DNA. Our results are consistent with the characteristics of membrane‐bound particles containing fetal DNA found in maternal plasma.
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关键词
apoptotic bodies,fetal DNA
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