Exposure of cells to static magnetic field accelerates loss of integrity of plasma membrane during apoptosis.
BACKGROUND
Much attention is being paid to the biologic effects of magnetic fields (MFs). Although MFs enhance tumorigenesis, they are neither mutagenic nor tumorigenic. The mechanism of their tumorigenic effect has not been elucidated.


METHODS
To investigate the effect of MFs on apoptosis in HL-60 cells, we exposed the cells to static MFs of 6 mT generated by a magnetic disk of known intensity. Apoptosis was triggered by the DNA topoisomerase I inhibitor, camptothecin (CPT). Activation of caspases in situ using the fluorochrome-labeled inhibitor (FLICA) method and determination of plasma membrane integrity by excluding propidium iodide (PI) were measured by both laser scanning cytometry (LSC) and flow cytometry (FC). LSC and FC identified cells at three sequential stages of their demise: early apoptosis (cells with activated caspases and PI negative); late apoptosis (cells with activated caspases but unable to exclude PI); secondary necrosis (cells with apoptotic morphology no longer stained with FLICA, not excluding PI).


RESULTS
MF alone did not induce any apoptogenic or necrogenic effect. CPT exposure led to the sequential appearance of apoptotic cells. In the presence of CPT and MF, the overall proportion of cells undergoing apoptosis was not significantly changed. However, we consistently observed a significant increase in the frequency of late apoptotic/necrotic cells when compared with samples treated with CPT alone (P < 0.001), as well as a decrease in the percentage of early apoptotic cells (P = 0.013). The data obtained by FC and LSC were consistent with each other, showing a similar phenomenon.


CONCLUSIONS
Whereas MF alone or with CPT did not affect overall cell viability, it accelerated the rate of cell transition from apoptosis to secondary necrosis after induction of apoptosis by the DNA-damaging agent, CPT. Modulation of the kinetics of the transition from apoptosis to secondary necrosis by MF in vivo may play a role in inflammation and tumorigenesis.
Teodori
Laura
L
Department of Biomedicine and Toxicology, UTS BIOTEC, ENEA-Casaccia, Rome, Italy.
Grabarek
Jerzy
J
Smolewski
Piotr
P
Ghibelli
Lina
L
Bergamaschi
Antonio
A
De Nicola
Milena
M
Darzynkiewicz
Zbigniew
Z
D017209:Apoptosis / Q000502:physiology / Q000528:radiation effects*; D002166:Camptothecin / Q000494:pharmacology; D002452:Cell Count; D002462:Cell Membrane / Q000528:radiation effects*; D002470:Cell Survival / Q000528:radiation effects; D005434:Flow Cytometry; D018922:HL-60 Cells / Q000473:pathology* / Q000528:radiation effects; D006801:Humans; D019044:Image Cytometry; D007938:Leukemia; D008280:Magnetics / Q000009:adverse effects*; D009336:Necrosis
D002166:Camptothecin