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Proceedings Paper

Changes in protein expression of U937 and Jurkat cells exposed to nanosecond pulsed electric fields
Author(s): Erick K. Moen; Caleb C. Roth; Caesar Cerna; Larry Estalck; Gerald Wilmink; Bennett L. Ibey
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Paper Abstract

Application of nanosecond pulsed electric fields (nsPEF) to various biological cell lines has been to shown to cause many diverse effects, including poration of the plasma membrane, depolarization of the mitochondrial membrane, blebbing, apoptosis, and intracellular calcium bursts. The underlying mechanism(s) responsible for these diverse responses are poorly understood. Of specific interest in this paper are the long-term effects of nsPEF on cellular processes, including the regulation of genes and production of proteins. Previous studies have reported transient activation of select signaling pathways involving mitogen-activated protein kinases (MAPKs), protein phosphorylation and downstream gene expression following nsPEF application. We hypothesize that nsPEF represents a unique stimulus that could be used to externally modulate cellular processes. To validate our hypothesis, we performed a series of cuvette-based exposures at 10 and 600ns pulse widths using a custom Blumlien line pulser system. We measured acute changes in the plasma membrane structure using flow cytometry by tracking phosphatidylserine externalization via FITC-Annexin V labeling and poration via propidium iodide uptake. We then compared these results to viability of the cells at 24 hours post exposure using MTT assay and changes in the MAPK family of proteins at 8 hours post-exposure using Luminex assay. By comparing exposures at 10 and 600ns duration, we found that most MAPK family-protein expression increased in Jurkat and U937 cell lines following exposure and compared well with drops in viability and changes in plasma membrane asymmetry. What proved interesting is that some MAPK family proteins (e.g. p53, STAT1), were expressed in one cell line, but not the other. This difference may point to an underlying mechanism for observed difference in cellular sensitivity to nsPEFinduced stresses.

Paper Details

Date Published: 23 February 2013
PDF: 9 pages
Proc. SPIE 8585, Terahertz and Ultrashort Electromagnetic Pulses for Biomedical Applications, 85850R (23 February 2013); doi: 10.1117/12.2005062
Show Author Affiliations
Erick K. Moen, Univ. of Southern California (United States)
Caleb C. Roth, General Dynamics Information Technology (United States)
Caesar Cerna, Air Force Research Lab. (United States)
Larry Estalck, Air Force Research Lab. (United States)
Gerald Wilmink, Air Force Research Lab. (United States)
Bennett L. Ibey, Air Force Research Lab. (United States)

Published in SPIE Proceedings Vol. 8585:
Terahertz and Ultrashort Electromagnetic Pulses for Biomedical Applications
Gerald J. Wilmink; Bennett L. Ibey, Editor(s)

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