nsPEF Application Research Articles

Evaluation of genetic response of mesenchymal stem cells to nanosecond pulsed electric fields by whole transcriptome sequencing.

Mesenchymal stem cells (MSCs) modulated by various exogenous signals have been applied extensively in regenerative medicine research. Notably, nanosecond pulsed electric fields (nsPEFs), characterized by short duration and high strength, significantly influence cell phenotypes and regulate MSCs differentiation via multiple pathways. Consequently, we used transcriptomics to study changes in messenger RNA (mRNA), long noncoding RNA (lncRNA), microRNA (miRNA), and circular RNA expression during nsPEFs application. To explore gene expression profiles and potential transcriptional regulatory mechanisms in MSCs pretreated with nsPEFs. The impact of nsPEFs on the MSCs transcriptome was investigated through whole transcriptome sequencing. MSCs were pretreated with 5-pulse nsPEFs (100 ns at 10 kV/cm, 1 Hz), followed by total RNA isolation. Each transcript was normalized by fragments per kilobase per million. Fold change and difference significance were applied to screen the differentially expressed genes (DEGs). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed to elucidate gene functions, complemented by quantitative polymerase chain reaction verification. In total, 263 DEGs were discovered, with 92 upregulated and 171 downregulated. DEGs were predominantly enriched in epithelial cell proliferation, osteoblast differentiation, mesenchymal cell differentiation, nuclear division, and wound healing. Regarding cellular components, DEGs are primarily involved in condensed chromosome, chromosomal region, actin cytoskeleton, and kinetochore. From aspect of molecular functions, DEGs are mainly involved in glycosaminoglycan binding, integrin binding, nuclear steroid receptor activity, cytoskeletal motor activity, and steroid binding. Quantitative real-time polymerase chain reaction confirmed targeted transcript regulation. Our systematic investigation of the wide-ranging transcriptional pattern modulated by nsPEFs revealed the differential expression of 263 mRNAs, 2 miRNAs, and 65 lncRNAs. Our study demonstrates that nsPEFs may affect stem cells through several signaling pathways, which are involved in vesicular transport, calcium ion transport, cytoskeleton, and cell differentiation.

Susceptibility of various human cancer cell lines to nanosecond and microsecond range electrochemotherapy: Feasibility of multi-drug cocktails

Electrochemotherapy (ECT) involves combining anticancer drugs with electroporation, which is induced by pulsed electric fields (PEFs), while the effects vary in effectiveness based on the specific parameters of the electrical pulses and susceptibility of the cells to a specific drug. In this work, we utilized conventional microsecond electroporation protocols (0.8 – 1.5 kV/cm × 100 μs × 8, 1 Hz) and the new modality of nanosecond pulses (4 and 8 kV/cm × 500 ns × 100, 1 kHz and 1 MHz), which are compressed into a high frequency burst. Sensitive and resistant lung, breast and ovarian human cancer cell lines were used in the study. In order to overcome drug-resistance, we have investigated the feasibility to use anticancer drug cocktails i.e., bleomycin and cisplatin combinations with metformin, vinorelbine and Dp44mT. The different susceptibility of various human cancer cells lines to electric pulses was determined, the efficacy of ECT was characterized and the type of cell death depending on the combinations of drugs was investigated. The results indicate that synergistic effects of PEFs with drug cocktails may be used to overcome drug-resistance in cancer, while the application of nsPEF provides more flexibility in parametric protocols and modulation of cancer cell death.

Application of Nanosecond Pulsed Electric Field and Autofluorescence Lifetime Microscopy of FAD in Lung Cells.

Exposure of nanosecond pulsed electric fields (nsPEFs) to live cells is an increasing research interest in biology and medicine. Despite extensive studies, a question still remains as to how effects of application of nsPEF on intracellular functions are different between cancerous cells and normal cells and how the difference can be detected. Herein, we have presented an approach of autofluorescence lifetime (AFL) microscopy of flavin adenine dinucleotide (FAD) to detect effects of application of nsPEF having 50 ns of a pulse width, nsPEF(50), on intracellular function in lung cancerous cells, A549 and H661, which show nsPEF(50)-induced apoptosis, and normal cells, MRC-5, in which the field effect is less or not induced. Then, the application of nsPEF(50) is shown to increase the lifetime of FAD autofluorescence in lung cancerous cells, whereas the electric field effects on the autofluorescence of FAD was not significant in normal healthy cells, which indicates that the lifetime measurements of FAD autofluorescence are applicable to detect the field-induced change in intracellular functions. Lifetime and intensity microscopic images of FAD autofluorescence in these lung cells were also acquired after exposure to the apoptosis-inducer staurosporine (STS). Then, it was found that the AFL of FAD became longer after exposure not only in the cancerous cells but also in the normal cells. These results indicate that nsPEF(50) applied to lung cells induced apoptotic cell death only in lung cancerous cells (H661 and A549) but not in lung normal cells (MRC-5), whereas STS induced apoptotic cell death both in lung cancerous cells and in lung normal cells. The lifetime microscopy of FAD autofluorescence is suggested to be very useful as a sensitive detection method of nsPEF-induced apoptotic cell death.

Fluorescence microscopic approach for detection of two different modes of breast cancer cell death induced by nanosecond pulsed electric field

Application of nanosecond pulsed electric field (nsPEF) to living systems is a fast-growing research area with great potential for cancer treatment. Here, we have demonstrated a cellular-level approach applying nsPEF having a pulse width of 50 ns and a frequency of 2 kHz to induce the change in intracellular function and dynamics for two subtypes of breast cancerous cells, that is, estrogen and progesterone-positive (ER+ and PR+) and human epidermal growth factor 2 (HER2) negative MCF-7 cells and highly aggressive drug resistance triple-negative MDA-MB-231 cells. Microscopic monitoring of the field effects in real-time or before and after the application of nsPEF indicated that nsPEF induced different field effects on intracellular function and dynamics in MCF-7 and MDA-MB-231 cells and activated two different modes of cell death, that is, apoptosis in MCF-7 cells and necrosis (or necroptosis) in MDA-MB-231 cells. These two different modes of nsPEF-induced cell death were confirmed based on the following microscopic measurements: 1) autofluorescence intensity and lifetime imaging of nicotinamide adenine dinucleotide (NADH); 2) production of reactive oxygen species (ROS); 3) change in mitochondrial membrane potential; 4) morphological alternation of cells; 5) imaging of phosphatidyl serine (PS) externalization by Annexin V; 6) cell viability. The present finding of nanosecond pulsed electric field-induced apoptosis and necrosis (or necroptosis) in subtypes of breast cancerous cells provides valuable information that the mechanism of field-induced cell death depends on the subtype of cancerous cells. It will be helpful for further development and optimization of field-induced cancer therapy.

Effects of nanosecond pulsed electric fields application on cancer cell and combination of anticancer drug

AbstractWith the goal of establishing a new cancer treatment method using nanosecond pulsed electric fields (nsPEFs), we investigated the effect of applying nsPEFs and anticancer drugs (adriamycin, ADM) to cancer cells. The surviving fraction of cancer cells was significantly reduced by nsPEFs application compared to the control. Compared to nsPEFs alone and ADM administration alone, a significant decrease of the surviving fraction was observed in combination of nsPEFs application and ADM administration. In addition, multiplying value of surviving fractions of nsPEFs application and ADM administration was calculated as a pseudo‐combination effect. The surviving fractions of the experimental combinational results of nsPEFs and ADM were lower than the pseudo‐combination results. Therefore, the synergistic effect of combination of nsPEFs and ADM was confirmed. Furthermore, the surviving fraction of ADM administration after nsPEFs application was significantly lower than that of ADM administration on 24 h before nsPEFs application. ADM action on cells would be effective owing to nanopores were formed by applying nsPEFs. It was confirmed that ADM and nsPEF induced apoptosis of cells from apoptosis and necrosis test using FACS.

Cancer specific apoptosis induced by electric field: A possible key mechanism in cell-competition and photodynamic action

Apoptosis induced by nanosecond pulsed electric field (nsPEF) has been demonstrated in non-small lung cancerous cells by the observation of field-induced change in cell morphology (swelling and blebbing), phosphatidylserine externalization and change in viability, whereas such field effect was harmless in normal lung cells. Lengthening of autofluorescence lifetime of nicotinamide adenine dinucleotide, decrease of mitochondrial membrane potential, activation of caspases, and generation of reactive oxygen species such as superoxide anion and singlet oxygen were clearly observed only in cancerous cells with application of nsPEF, indicating that the field-induced apoptotic cell death is initiated intrinsically in mitochondria. The finding implicated that electric field effect may be a key mechanism in “cell-competition” between normal and cancerous cells to facilitate the elimination of transformed cancerous cells from surrounding normal cells, and a new insight in “photodynamic action” to kill cancerous cells, which is a root of photodynamic therapy, has been provided.

Exploring the Conformational Changes Induced by Nanosecond Pulsed Electric Fields on the Voltage Sensing Domain of a Ca2+ Channel.

Nanosecond Pulsed Electric Field (nsPEF or Nano Pulsed Stimulation, NPS) is a technology that delivers a series of pulses of high-voltage electric fields during a short period of time, in the order of nanoseconds. The main consequence of nsPEF upon cells is the formation of nanopores, which is followed by the gating of ionic channels. Literature is conclusive in that the physiological mechanisms governing ion channel gating occur in the order of milliseconds. Hence, understanding how these channels can be activated by a nsPEF would be an important step in order to conciliate fundamental biophysical knowledge with improved nsPEF applications. To get insights on both the kinetics and thermodynamics of ion channel gating induced by nsPEF, in this work, we simulated the Voltage Sensing Domain (VSD) of a voltage-gated Ca channel, inserted in phospholipidic membranes with different concentrations of cholesterol. We studied the conformational changes of the VSD under a nsPEF mimicked by the application of a continuous electric field lasting 50 ns with different intensities as an approach to reveal novel mechanisms leading to ion channel gating in such short timescales. Our results show that using a membrane with high cholesterol content, under an nsPEF of 50 ns and = 0.2 V/nm, the VSD undergoes major conformational changes. As a whole, our work supports the notion that membrane composition may act as an allosteric regulator, specifically cholesterol content, which is fundamental for the response of the VSD to an external electric field. Moreover, changes on the VSD structure suggest that the gating of voltage-gated Ca channels by a nsPEF may be due to major conformational changes elicited in response to the external electric field. Finally, the VSD/cholesterol-bilayer under an nsPEF of 50 ns and = 0.2 V/nm elicits a pore formation across the VSD suggesting a new non-reported effect of nsPEF into cells, which can be called a “protein mediated electroporation”.

Nanosecond pulsed electric fields impair viability and mucin expression in mucinous colorectal carcinoma cell

Nanosecond pulsed electric fields (nsPEFs) are a non-thermal technology that can induce a myriad of biological responses and changes in cellular physiology. nsPEFs have gained significant attention as a novel cancer therapy. However, studies investigating the application of nsPEF in mucinous carcinomas are scarce. In this study, we explored several biological responses in two mucinous colorectal adenocarcinoma cell lines, LS 174T and HT-29, to nsPEF treatment. We determined the overall cell survival and viability rates following nsPEF treatment using CCK-8 and colony formation assays. We measured the intracellular effects of nsPEF treatment by analyzing cell cycle distribution, cell apoptosis and mitochondrial potential. We also analyzed mucin production at both mRNA and protein levels. Our results showed that nsPEF treatment significantly reduced mucinous cell viability in a dose-dependent manner. nsPEF treatment increased cell cycles arrest at G0/G1 while the proportion of G2/M cells gradually decreased. Cell apoptosis increased following nsPEF treatment with a clear loss in mitochondrial membrane potential. Furthermore, the protein expression of functional mucin family members decreased after nsPEF treatment. In conclusion, nsPEF treatment reduced MCRC cell viability, cell proliferation, and mucin protein production while promoted apoptosis. Our work is a pilot study that projects some insights into the potential clinical applications of nsPEFs in treating mucinous colorectal carcinoma.

High-voltage 10 ns delayed paired or bipolar pulses for in vitro bioelectric experiments

Recent studies proved that classical bio-effects induced by nanosecond pulsed electric field (nsPEF) can be reduced by the delivery of a negative polarity pulse generated immediately after a positive polarity pulse. This phenomenon is known as “bipolar cancellation” and it was reported for a wide range of bipolar pulses with pulse duration from 2 ns to 900 ns. On the contrary, paired pulses, i.e., two identical pulses with the same polarity, increased traditional nsPEF outcomes. Herein, we propose a novel robust and flexible generator, based on the frozen-wave concept, able to produce a broad range of pulses with the duration of 10 ns and delay between 17 and 360 ns. Numerical simulations and experimental measurements were performed to fully characterize the proposed generator. YO-PROTM-1 uptake was investigated in the U87-MG human glioblastoma cell line as a marker of membrane permeabilization in response to 10 ns, 11.5MV/m nsPEF. Our results showed that bipolar cancellation occurred for delays of 0–30 ns and decreased as a function of the interphase interval. In addition, we observed that cellular response following the application of paired nsPEF was more than two-fold compared to the unipolar pulse response and was independent from the interphase interval.

Effect of nanosecond pulsed electric field treatment on cell proliferation of microalgae

Photoautotrophic microalgae based biorefinery concepts are currently not competitive compared to other established production systems. Therefore, innovative upstream processes need to be developed to increase the competitiveness of photoautotrophic microalgae biorefinery concepts. Abiotic sub-lethal stress induction via nanosecond pulsed electric field (nsPEF) treatment might be a viable process to increase the efficiency of photoautotrophic microalgae cultivation. In this work, an increased cell growth after nsPEF treatment was observable. Application of nsPEF to highly proliferating cells in a repetitive process resulted in a statistical significant increase in cell growth (p = 0.009). The effect was most pronounced after five days wherefore cellular structures and processes were analyzed to reveal a possible mechanism. Within this work, a protocol for increased cell proliferation with a possible mechanism was derived, which improves competitiveness of photoautotrophic microalgae biorefineries in the future. However, based on the derived mechanism, the results are also relevant for other microorganisms.

Ns Pulsed Electric Field-Induced Action Potentials in the Circuital Model of an Axon.

Pulsed electric fields with duration in the sub- and ns time scale (nsPEFs) increase the permeability of cell membranes, enabling the transport of normally impermeant molecules into or out of the cell (electroporation). Such effect is associated to intracellular alterations and indicates nsPEFs as a new stimulus to modulate cell functions. In particular, studies dealing with the application of nsPEFs to excitable cells suggest their use for the stimulation/inhibition of cell excitation. In this paper, the circuital model per surface unit of the plasma membrane of an axon was developed to implement the Hodgkin and Huxley equations, describing the action potential activation process. For the first time, a power electronics circuital simulator was adopted. The model was first validated with conventional microsecond stimuli, and then it was employed to identify the conditions for cell excitation by nsPEFs. The results demonstrated the possibility of electrostimulation by nsPEFs at depolarization levels far below those required for inducing electroporation, and with ionic current dynamics similar to that induced by conventional stimuli, confirming recent experimental findings. Moreover, by using a power electronics tool, easier integration of the cell modeling with the design and optimization of pulse generation systems can be gained.

Effects of Nanosecond Pulsed Electric Field on Intracellular NADH Autofluorescence: A Comparison between Normal and Cancer Cells.

Intracellular fluorescence lifetime and intensity images of the endogenous fluorophore of nicotinamide adenine dinucleotide (NADH) have been observed before and after application of nanosecond pulsed electric field (nsPEF) in normal and cancer cells, that is, in Wistar-King-Aptekman rat fetus fibroblast (WFB) cells and W31 cells, which are the malignant transformed cells from WFB. The application of nsPEF induces a change both in intensity and lifetime of NADH, indicating that the intracellular function is affected by application of nsPEF in both normal and cancer cells. The application of nsPEF induces an increase in the fluorescence lifetime of NADH and a morphological change, which is attributed to the induction of apoptosis by nsPEF. The field effect on the intensity and lifetime clearly depends on the pulse width, and magnitude of the field-induced increase in the fluorescence lifetime of NADH has a tendency to increase with a decreasing pulse width. It is also found that apoptosis can be induced only in cancer cells using a suitable nsPEF, showing a possibility that ultrashort pulsed electric field is applicable for drug-free cancer therapy.

Nanosecond pulsed electric field induced changes in cell surface charge density

This study reports that the surface charge density changes in Jurkat cells with the application of single 60 nanosecond pulse electric fields, using atomic force microscopy. Using an atomic force microscope tip and Jurkat cells on silica in a 0.01M KCl ionic concentration, we were able to measure the interfacial forces, while also predicting surface charge densities of both Jurkat cell and silica surfaces. The most important finding is that the pulsing conditions varyingly reduced the cells’ surface charge density. This offers a novel way in which to examine cellular effects of pulsed electric fields that may lead to the identification of unique mechanical responses. Compared to a single low field strength NsPEF (15kV/cm) application, exposure of Jurkat cells to a single high field strength NsPEF (60kV/cm) resulted in a further reduction in charge density and major morphological changes. The structural, physical, and chemical properties of biological cells immensely influence their electrostatic force; we were able to investigate this through the use of atomic force microscopy by measuring the surface forces between the AFM's tip and the Jurkat cells under different pulsing conditions as well as the interfacial forces in ionic concentrations.

A new mechanism for efficient hydrocarbon electro-extraction from Botryococcus braunii

BackgroundRecent understanding that specific algae have high hydrocarbon production potential has attracted considerable attention. Botryococcus braunii is a microalga with an extracellular hydrocarbon matrix, which makes it an appropriate green energy source.ResultsThis study focuses on extracting oil from the microalgae matrix rather than the cells, eliminating the need for an excessive electric field to create electro-permeabilization. In such a way, technical limitations due to high extraction energy and cost can be overcome. Here, nanosecond pulsed electric fields (nsPEF) with 80 ns duration and 20–65 kV/cm electric fields were applied. To understand the extraction mechanism, the structure of the algae was accurately studied under fluorescence microscope; extraction was quantified using image analysis; quality of extraction was examined by thin-layer chromatography (TLC); and the cell/matrix separation was observed real-time under a microscope during nsPEF application. Furthermore, optimization was carried out by screening values of electric fields, pulse repetition frequencies, and energy spent.ConclusionsThe results offer a novel method applicable for fast and continues hydrocarbon extraction process at low energy cost.

Calcium-independent disruption of microtubule dynamics by nanosecond pulsed electric fields in U87 human glioblastoma cells

High powered, nanosecond duration, pulsed electric fields (nsPEF) cause cell death by a mechanism that is not fully understood and have been proposed as a targeted cancer therapy. Numerous chemotherapeutics work by disrupting microtubules. As microtubules are affected by electrical fields, this study looks at the possibility of disrupting them electrically with nsPEF. Human glioblastoma cells (U87-MG) treated with 100, 10 ns, 44 kV/cm pulses at a frequency of 10 Hz showed a breakdown of their interphase microtubule network that was accompanied by a reduction in the number of growing microtubules. This effect is temporally linked to loss of mitochondrial membrane potential and independent of cellular swelling and calcium influx, two factors that disrupt microtubule growth dynamics. Super-resolution microscopy revealed microtubule buckling and breaking as a result of nsPEF application, suggesting that nsPEF may act directly on microtubules.

Effects of Nanosecond Pulsed Electric Fields on the Intracellular Function of HeLa Cells As Revealed by NADH Autofluorescence Microscopy.

The fluorescence lifetime of the endogenous fluorophore of reduced nicotinamide adenine dinucleotide (NADH) in HeLa cells is affected by the application of nanosecond pulsed electric fields (nsPEFs). In this study, we found that after nsPEF application, the fluorescence lifetime became longer and then decreased in a stepwise manner upon further application, irrespective of the pulse width in the range of 10–50 ns. This application time dependence of the NADH fluorescence lifetime is very similar to the time-lapse dependence of the NADH fluorescence lifetime following the addition of an apoptosis inducer, staurosporine. These results, as well as the membrane swelling and blebbing after the application of nsPEFs, indicate that apoptosis is also induced by the application of nsPEFs in HeLa cells. In contrast to the lifetime, the fluorescence intensity remarkably depended on the pulse width of the applied nsPEF. When the pulse width was as large as 50 ns, the intensity monotonically increased and was distributed over the entire cell as the application duration became longer. As the pulse width of the applied electric field became smaller, the magnitude of the field-induced increase in NADH fluorescence intensity decreased; the intensity was reduced by the electric field when the pulse width was as small as 10 ns. These results suggest that the mechanism of electric-field-induced apoptosis depends on the pulse width of the applied nsPEF.

Ablation of Myocardial Tissue With Nanosecond Pulsed Electric Fields

BackgroundAblation of cardiac tissue is an essential tool for the treatment of arrhythmias, particularly of atrial fibrillation, atrial flutter, and ventricular tachycardia. Current ablation technologies suffer from substantial recurrence rates, thermal side effects, and long procedure times. We demonstrate that ablation with nanosecond pulsed electric fields (nsPEFs) can potentially overcome these limitations.MethodsWe used optical mapping to monitor electrical activity in Langendorff-perfused New Zealand rabbit hearts (n = 12). We repeatedly inserted two shock electrodes, spaced 2–4 mm apart, into the ventricles (through the entire wall) and applied nanosecond pulsed electric fields (nsPEF) (5–20 kV/cm, 350 ns duration, at varying pulse numbers and frequencies) to create linear lesions of 12–18 mm length. Hearts were stained either with tetrazolium chloride (TTC) or propidium iodide (PI) to determine the extent of ablation. Some stained lesions were sectioned to obtain the three-dimensional geometry of the ablated volume.ResultsIn all animals (12/12), we were able to create nonconducting lesions with less than 2 seconds of nsPEF application per site and minimal heating (< 0.2°C) of the tissue. The geometry of the ablated volume was smoother and more uniform throughout the wall than typical for RF ablation. The width of the lesions could be controlled up to 6 mm via the electrode spacing and the shock parameters.ConclusionsAblation with nsPEFs is a promising alternative to radiofrequency (RF) ablation of AF. It may dramatically reduce procedure times and produce more consistent lesion thickness than RF ablation.

Nanosecond pulsed electric fields (nsPEFs) impact and enhanced Photofrin II® delivery in photodynamic reaction in cancer and normal cells

BackgroundNanosecond pulsed electric field (nsPEF) treatment is a new anti-cancer strategy with ultrashort pulse duration and high intensity of electric fields. The application of nsPEFs affects all intra- and extracellular membranes and independently initiates the process of apoptosis within cancer cells, leading the tumor to slowly auto-destruct without the use of toxic drugs. MethodsThis study involves cells of gastric adenocarcinoma (EPG85-257P and EPG85-257RDB), metastatic melanoma (Me45), epidermal cancer (A431), normal keratinocytes (HaCaT), and macrophages (P388/D1). The influence of nanosecond pulses on the cellular structure and cellular proliferation was evaluated. The effect of nsPEF was determined by MTT and clonogenic assays and the efficiency was monitored by following the propidium iodide and Photofrin II® uptake using FACS analysis. The cell membranes state was visualized with DHCC marker. ResultsnsPEFs (up to 60kV/cm) induced significant decrease of cellular viability in all cancer cells except the A431 cell line. Photodynamic reactions combined with nsPEFs induced the highest decrease of cellular viability in both gastric cell lines and skin derived cancer cells. Normal (HaCaT and P388/D1) cells were in contrary not significantly affected. Propidium iodide and Photofrin II® uptake, used as markers of membrane permeabilization, were the most efficient in gastric cells. Finally, the most disturbed morphology was observed in the latter. ConclusionsThis is the first attempt of combining nsPEF with photodynamic reaction using Ph II® for selective destruction of cancer cells. The results indicate the potential of nsPEF for inducing cytotoxicity mainly in adenocarcinoma cells, while combined with Photofrin II® and irradiation.

Effects of Burst Electromagnetic Wave Focusing and Their Combination With Nanosecond Pulsed Electric Fields on HeLa S3 Cells

Burst electromagnetic waves focusing, nanosecond pulsed electric fields (nsPEF), and their combinations were applied to HeLa S3 cells to investigate their respective biological effects. The research was motivated by its application for cancer therapy. Burst electromagnetic waves, characterized by high-frequency (800 MHz) and narrow-band signals, were applied through a focusing system consisting of an ellipsoidal reflector and a radiating antenna. Burst widths of electromagnetic waves were adjusted to 1 and 5 ms. For nsPEF exposure, a magnetic pulse power modulator with an 80-ns pulsewidth and repetition frequencies of 0.5 and 1 pulse per second was employed. Cervical cancer cells (HeLa S3) subjected to electric fields were cultured in an incubator for 24 h and stained using propidium iodide to monitor the cell death rate. Sole application of burst electromagnetic waves was found not to change the cancer cell survival, while sole application of nsPEF suppressed the cells growth. The combination of nsPEF and focused burst electromagnetic wave was found to be very effective in producing maximum cell death rate. The results show the importance of the exposure combination component, as a first step, to design a more efficient and less-invasive cancer therapy system.

Synergistic effect of nanosecond pulsed electric field combined with low-dose of pingyangmycin on salivary adenoid cystic carcinoma

Adenoid cystic carcinoma (ACC) is one of the most common malignant neoplasms in salivary glands. To evaluate the therapeutic effects of nanosecond pulsed electric field (nsPEF) combined with pingyangmycin (PYM) on salivary gland adenoid cystic carcinoma (SACC), ACC high metastatic cell line (SACC-LM) and low metastatic cell line (SACC‑83) were tested by CCK-8 assay, cell clonogenic assay, flow cytometry and Transwell assay. Extracellular matrix metalloproteinase inducer (EMMPRIN) expression was tested by western blotting to verify the synergistic mechanism of nsPEF and PYM. The results showed that nsPEF inhibited the cell proliferation of both cell lines, and the inhibitory effect was strongly associated with time and electrical field strength. Moreover, PYM combined with nsPEF may enhance the suppression effect significantly, even at a very low dose (0.01 µg/ml). The synergistic effects may contribute to the downregulation of EMMPRIN expression resulting from the application of nsPEF. For SACC, nsPEF combined with chemotherapy agents may be a valuable strategy not only to improve the treatment effect and prognosis, but also to reduce the side-effects of chemotherapy.