9 research outputs found
Metabolomics Using 1H-NMR of apoptosis and necrosis in HL60 leukemia cells: Differences between the two types of cell death and independence from the stimulus of apoptotis used.
High-resolution proton nuclear magnetic resonance (1HNMR)
spectroscopy was used to examine and compare the
metabolic variations that occur in cells of the HL60 promyelocytic
leukemia cell line after induction of apoptosis by ionizing
radiation and the antineoplastic drug doxorubicin as
well as after induction of necrosis by heating. Apoptosis and
necrosis were confirmed by fluorescence microscopy using the
chromatin stain Hoechst 33258, agarose gel electrophoresis of
DNA, and determination of caspase 3 enzymatic activity. The
1H-NMR experiments revealed that the spectra of both samples
containing apoptotic cells were characterized by the same
trend of several important metabolites. Specifically, an increase
in CH2 and CH3 mobile lipids, principally of CH2, decreases
in glutamine and glutamate, choline-containing metabolites,
taurine and reduced glutathione were observed. By
contrast, the sample containing necrotic cells presented a completely
different profile of 1H-NMR metabolites since it was
characterized by a significant increase in all the metabolites
examined, with the exception of CH2 mobile lipids, which remain
unchanged, and reduced glutathione, which decreased.
The results suggest that variations in 1H-NMR metabolites are
specific to apoptosis independent of the physical or chemical
nature of the stimulus used to induce this mode of cell death,
while cells dying from necrosis are characterized by a completely
different behavior of the same metabolites