Detection of drug-induced apoptosis and necrosis in human cervical carcinoma cells using 1H NMR spectroscopy.
Apoptosis and necrosis need to be differentiated in order to distinguish drug-induced cell death from spontaneous cell death due to hypoxia. The ability to differentiate between these two modes of cell death, especially at an early stage in the process, could have a significant impact on accessing the outcome of anticancer drug therapy in the clinic. Nuclear magnetic resonance spectroscopy was used to distinguish apoptosis from necrosis in human cervical carcinoma (HeLa) cells. Apoptosis was induced by treatment with the topoisomerase II inhibitor etoposide, whereas necrosis was induced by the use of ethacrynic acid or cytochalasin B. We found that the intensity of the methylene resonance increases significantly as early as 6 h after the onset of apoptosis, but that no such changes occur during necrosis. The spectral intensity ratio of the methylene to methyl resonances also shows a high correlation with the percentage of apoptotic cells in the sample (r2=0.965, P<0.003).
Bezabeh
T
T
Institute for Biodiagnostics, National Research Council, Winnipeg, Manitoba, R3B 1Y6 Canada. Tedros.Bezabeh@nrc.ca
Mowat
M R
MR
Jarolim
L
L
Greenberg
A H
AH
Smith
I C
IC
D017209:Apoptosis / Q000187:drug effects*; D003571:Cytochalasin B / Q000494:pharmacology; D004976:Ethacrynic Acid / Q000494:pharmacology; D005260:Female; D006367:HeLa Cells; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009336:Necrosis / Q000139:chemically induced; D011522:Protons; D002583:Uterine Cervical Neoplasms / Q000188:drug therapy / Q000473:pathology*
D011522:Protons; D003571:Cytochalasin B; D004976:Ethacrynic Acid