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Quantitative analysis of apoptotic cell death using proton nuclear magnetic
resonance spectroscopy
FG Blankenberg, PD Katsikis, RW Storrs, C Beaulieu, D Spielman, JY Chen, L Naumovski and JF Tait
Department of Radiology, Stanford University School of Medicine, CA, USA.
Quantification of apoptotic cell death in vivo has become an important area
of investigation in patients with acute lymphoblastic leukemia (ALL). We
have devised a noninvasive analytical method to estimate the percentage of
apoptotic lymphoblasts in doxorubicin-treated Jurkat T- cell ALL cultures,
using proton nuclear magnetic resonance spectroscopy (1H NMR). We have
found that the ratio of the methylene (CH2) resonance (at 1.3 ppm) to the
methyl (CH3) resonance (at 0.9 ppm) signal intensity, as observed by 1H
NMR, is directly proportional to the percentage of apoptotic lymphoblasts
in vitro. The correlation between the CH2/CH3 signal intensity ratio and
the percentage of apoptotic lymphoblasts was optimal 24 to 28 hours after
doxorubicin treatment (r2 = .947, N = 27 samples). There was also a direct
temporal relationship between an increase in the CH2/CH3 signal intensity
ratio and the onset of apoptosis as detected by nuclear morphologic
analysis, fluorescein- annexin V flow cytometry, and DNA gel
electrophoresis. Thin-layer chromatography confirmed that a dynamic and/or
compositional change of the plasma membrane, rather than increases in
lipase activity or fatty acid production, appears to account for the
increase in the CH2/CH3 signal intensity ratio during apoptosis. 1H NMR may
have clinical utility for the early noninvasive assessment of
chemotherapeutic efficacy in patients with ALL.
Volume 89,
Issue 10,
pp. 3778-3786,
05/15/1997
Copyright © 1997 by The American Society of Hematology
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