Magnetic resonance detects metabolic changes associated with chemotherapy-induced apoptosis.
Apoptosis was induced by treating L1210 leukaemia cells with mechlorethamine, and SW620 colorectal cells with doxorubicin. The onset and progression of apoptosis were monitored by assessing caspase activation, mitochondrial transmembrane potential, phosphatidylserine externalization, DNA fragmentation and cell morphology. In parallel, 31P magnetic resonance (MR) spectra of cell extracts were recorded. In L1210 cells, caspase activation was detected at 4 h. By 3 h, the MR spectra showed a steady decrease in NTP and NAD, and a significant build-up of fructose 1,6-bisphosphate (F-1,6-P) dihydroxyacetonephosphate and glycerol-3-phosphate, indicating modulation of glycolysis. Treatment with iodoacetate also induced a build-up of F-1,6-P, while preincubation with two poly(ADP-ribose) polymerase inhibitors, 3-aminobenzamide and nicotinamide, prevented the drop in NAD and the build-up of glycolytic intermediates. This suggested that our results were due to inhibition of glyceraldehyde-3-phosphate dehydrogenase, possibly as a consequence of NAD depletion following poly(ADP-ribose) polymerase activation. Doxorubicin treatment of the adherent SW620 cells caused cells committed to apoptosis to detach. F-1,6-P was observed in detached cells, but not in treated cells that remained attached. This indicated that our observations were not cell line- or treatment-specific, but were correlated with the appearance of apoptotic cells following drug treatment. The 31P MR spectrum of tumours responding to chemotherapy could be modulated by similar effects.
Ronen
S M
SM
Cancer Research Campaign, Clinical Magnetic Resonance Research Group, Institute of Cancer Research, Sutton, Surrey, UK.
DiStefano
F
F
McCoy
C L
CL
Robertson
D
D
Smith
T A
TA
Al-Saffar
N M
NM
Titley
J
J
Cunningham
D C
DC
Griffiths
J R
JR
Leach
M O
MO
Clarke
P A
PA
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis; D001549:Benzamides / Q000494:pharmacology; D020169:Caspases / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D004099:Dihydroxyacetone Phosphate / Q000378:metabolism; D004317:Doxorubicin / Q000494:pharmacology; D005635:Fructosediphosphates / Q000378:metabolism; D005994:Glycerophosphates / Q000378:metabolism; D006801:Humans; D007461:Iodoacetates / Q000494:pharmacology; D007939:Leukemia L1210 / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D009682:Magnetic Resonance Spectroscopy; D008466:Mechlorethamine / Q000494:pharmacology; D009243:NAD / Q000378:metabolism; D009536:Niacinamide / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D001549:Benzamides; D005635:Fructosediphosphates; D005994:Glycerophosphates; D007461:Iodoacetates; D009243:NAD; D009536:Niacinamide; D008466:Mechlorethamine; D004099:Dihydroxyacetone Phosphate; D004317:Doxorubicin; C025160:3-aminobenzamide; C029620:alpha-glycerophosphoric acid; D020169:Caspases; C029063:fructose-1,6-diphosphate