Evaluation of the Genetic Response of U937 and Jurkat Cells to 10-Nanosecond Electrical Pulses (nsEP)

Roth, Caleb C.; Glickman, Randolph D.; Tolstykh, Gleb P.; Estlack, Larry E.; Moen, Erick; Echchgadda, Ibtissam; Beier, Hope T.; Barnes, Ronald Jr A; Ibey, Bennett L.

doi:10.1371/journal.pone.0154555

Cited by 15 publications

(17 citation statements)

References 60 publications

(57 reference statements)

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“…We found that these shock waves were most likely not directly responsible for nanoporation; however, we hypothesized that they might be strong enough to elicit a mechanical stress response. We performed a microarray analysis on both Jurkat and U937 cells exposed to nsEP and identified genetic markers of mechanical stress [31] . However, these cell types are not generally used in mechanobiological research; therefore the experiments were repeated with cells used in mechanobiology.…”

Section: Discussionmentioning

confidence: 99%

“…Cells were counted using the Countess® Cell Counter from Life Technologies (Grand Island, NY) and the final concentration was adjusted to 1200 cells/μL in complete growth medium. The cells were then aliquoted into 1 mm gap (150 μL volume) electroporation cuvettes (VWR, Radnor, PA) and were exposed to either nsEP or they were SHAM exposed (both in complete growth medium) [31] . SHAM and nsEP exposures occurred in a random fashion.…”

Section: Methodsmentioning

confidence: 99%

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Adult human dermal fibroblasts exposed to nanosecond electrical pulses exhibit genetic biomarkers of mechanical stress

Roth

Glickman

Martens³

et al. 2017

Biochemistry and Biophysics Reports

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BackgroundExposure of cells to very short (<1 µs) electric pulses in the megavolt/meter range have been shown to cause a multitude of effects, both physical and molecular in nature. Physically, nanosecond electrical pulses (nsEP) can cause disruption of the plasma membrane, cellular swelling, shrinking and blebbing. Molecularly, nsEP have been shown to activate signaling pathways, produce oxidative stress, stimulate hormone secretion and induce both apoptotic and necrotic death. We hypothesize that studying the genetic response of primary human dermal fibroblasts exposed to nsEP, will gain insight into the molecular mechanism(s) either activated directly by nsEP, or indirectly through electrophysiology interactions.MethodsMicroarray analysis in conjunction with quantitative real time polymerase chain reaction (qRT-PCR) was used to screen and validate genes selectively upregulated in response to nsEP exposure.ResultsExpression profiles of 486 genes were found to be significantly changed by nsEP exposure. 50% of the top 20 responding genes coded for proteins located in two distinct cellular locations, the plasma membrane and the nucleus. Further analysis of five of the top 20 upregulated genes indicated that the HDFa cells’ response to nsEP exposure included many elements of a mechanical stress response.ConclusionsWe found that several genes, some of which are mechanosensitive, were selectively upregulated due to nsEP exposure. This genetic response appears to be a primary response to the stimuli and not a secondary response to cellular swelling.General significanceThis work provides strong evidence that cells exposed to nsEP interpret the insult as a mechanical stress.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Adult human dermal fibroblasts exposed to nanosecond electrical pulses exhibit genetic biomarkers of mechanical stress

Roth

Glickman

Martens³

et al. 2017

Biochemistry and Biophysics Reports

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“…Additionally, we investigated the mRNA expression of Caspase‐4 as a possible initiator of intrinsic apoptosis and utilized JUN (Jun Proto‐Oncogene, AP‐1 Transcription Factor Subunit) as a positive control in our experiments, as JUN is known to be upregulated post‐nsPEF exposure in U937 and Jurkat cell lines [Roth et al, ].…”

Section: Introductionmentioning

confidence: 99%

Nanosecond pulsed electric field exposure does not induce the unfolded protein response in adult human dermal fibroblasts

Martens

Roth

Ibey

2018

Bioelectromagnetics

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Cell-circuit models have suggested that nanosecond pulsed electric fields (nsPEFs) can disrupt intracellular membranes including endoplasmic reticulum (ER), mitochondria, and/or nucleus thereby inducing intrinsic apoptotic pathways. Therefore, we hypothesized that the unfolded protein response (UPR) would be activated, due to the fluctuations of ionic concentrations, upon poration of the ER membrane. Quantitative real-time polymerase chain reaction was utilized to measure changes in messenger RNA (mRNA) expression of specific ER stress genes in adult human dermal fibroblast (HDFa) cells treated with tunicamycin (TM) (known ER stress inducer) and cells exposed to nsPEFs (100, 10-ns pulses at 150 kV/cm delivered at a repetition rate of 1 Hz). For HDFa cells, results showed time-dependent UPR activation to TM; however, when HDFa cells were exposed to nsPEFs, no significant changes in mRNA expression of ER stress genes, and/or caspase gene were observed. These results indicate that although cell death can be observed under these exposure parameters, it is most likely not initiated through activation of the UPR. Bioelectromagnetics. 2018;39:491-499, 2018. Published 2018. This article is a U.S. Government work and is in the public domain in the USA.

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“…Transcriptome analysis, in our study, had the unique property of including the immediate, early (4 h), and late response (24 h) of cells to nsPEF exposure, in contrast to previously published studies analysing the differential gene expression of the immediate (at 0.5 and 1 h) [ 77 ] and early (at 4 h) cell responses [ 78 , 79 ]. This allowed us to trace the nsPEF-related variations in gene expression for the first time over a more extended period.…”

Section: Discussionmentioning

confidence: 95%

Nanosecond Pulsed Electric Field Only Transiently Affects the Cellular and Molecular Processes of Leydig Cells

Kasprzycka

Trębińska-Stryjewska

Lewandowski

et al. 2021

IJMS

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The purpose of this study was to verify whether the nanosecond pulsed electric field, not eliciting thermal effects, permanently changes the molecular processes and gene expression of Leydig TM3 cells. The cells were exposed to a moderate electric field (80 quasi-rectangular shape pulses, 60 ns pulse width, and an electric field of 14 kV/cm). The putative disturbances were recorded over 24 h. After exposure to the nanosecond pulsed electric field, a 19% increase in cell diameter, a loss of microvilli, and a 70% reduction in cell adhesion were observed. Some cells showed the nonapoptotic externalization of phosphatidylserine through the pores in the plasma membrane. The cell proportion in the subG1 phase increased by 8% at the expense of the S and G2/M phases, and the DNA was fragmented in a small proportion of the cells. The membrane mitochondrial potential and superoxide content decreased by 37% and 23%, respectively. Microarray’s transcriptome analysis demonstrated a negative transient effect on the expression of genes involved in oxidative phosphorylation, DNA repair, cell proliferation, and the overexpression of plasma membrane proteins. We conclude that nanosecond pulsed electric field affected the physiology and gene expression of TM3 cells transiently, with a noticeable heterogeneity of cellular responses.

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Evaluation of the Genetic Response of U937 and Jurkat Cells to 10-Nanosecond Electrical Pulses (nsEP)

Cited by 15 publications

References 60 publications

Adult human dermal fibroblasts exposed to nanosecond electrical pulses exhibit genetic biomarkers of mechanical stress

Adult human dermal fibroblasts exposed to nanosecond electrical pulses exhibit genetic biomarkers of mechanical stress

Nanosecond pulsed electric field exposure does not induce the unfolded protein response in adult human dermal fibroblasts

Nanosecond Pulsed Electric Field Only Transiently Affects the Cellular and Molecular Processes of Leydig Cells

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