Nanosecond pulsed electric fields (nsPEF) induce direct electric field effects and biological effects on human colon carcinoma cells.
Nanosecond pulsed electric fields (nsPEFs) are ultrashort pulses with high electric field intensity (kV/cm) and high power (megawatts), but low energy density (mJ/cc). To determine roles for p53 in response to nsPEFs, HCT116 cells (p53+/+ and p53-/-) were exposed to nsPEF and analyzed for membrane integrity, phosphatidylserine externalization, caspase activation, and cell survival. Decreasing plasma membrane effects were observed in both HCT116p53+/+ and p53-/- cells with decreasing pulse durations and/or decreasing electric fields. However, addition of ethidium homodimer-1 and Annexin-V-FITC post-pulse demonstrated greater fluorescence in p53-/- versus p53+/+ cells, suggesting a postpulse p53-dependent biological effect at the plasma membrane. Caspase activity was significantly higher than nonpulsed cells only in the p53-/- cells. HCT116 cells exhibited greater survival in response to nsPEFs than HL-60 and Jurkat cells, but survival was more evident for HCT116p53+/+ cells than for HCT116p53-/- cells. These results indicate that nsPEF effects on HCT116 cells include (1) apparent direct electric field effects, (2) biological effects that are p53-dependent and p53-independent, (3) actions on mechanisms that originate at the plasma membranes and at intracellular structures, and (4) an apparent p53 protective effect. NsPEF applications provide a means to explore intracellular structures and functions that can reveal mechanisms in health and disease.
Hall
Emily H
EH
Center for Pediatric Research, Eastern Virginia Medical School, Children's Hospital of the King's Daughters, Norfolk, Virginia 23510, USA.
Schoenbach
Karl H
KH
Beebe
Stephen J
SJ
D017304:Annexin A5; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism* / Q000528:radiation effects*; D002467:Cell Nucleus / Q000378:metabolism / Q000528:radiation effects; D002470:Cell Survival / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D004574:Electromagnetic Fields; D004789:Enzyme Activation; D004996:Ethidium / Q000031:analogs & derivatives* / Q000494:pharmacology; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins / Q000378:metabolism; D045325:HCT116 Cells; D018922:HL-60 Cells; D006801:Humans; D006834:Hydrazines; D007364:Intercalating Agents / Q000494:pharmacology; D019169:Jurkat Cells; D018613:Microscopy, Confocal; D010718:Phosphatidylserines / Q000032:analysis; D013997:Time Factors; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism* / Q000528:radiation effects*
C417561:Alexa 488 hydrazide; D017304:Annexin A5; D061945:Caspase Inhibitors; D005456:Fluorescent Dyes; D006834:Hydrazines; D007364:Intercalating Agents; D010718:Phosphatidylserines; D016159:Tumor Suppressor Protein p53; D049452:Green Fluorescent Proteins; C018533:ethidium homodimer; D020169:Caspases; D004996:Ethidium; D016650:Fluorescein-5-isothiocyanate