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

Awasthi, Kamlesh; Li, Si-Pei; Zhu, Chaoyuan; Hsu, Hsin-Yun; Ohta, Nobuhiro

doi:10.1016/j.snb.2022.133199

Cited by 7 publications

(3 citation statements)

References 51 publications

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“…The direct interaction of nsPEF to inner mitochondria was suggested to stop the ATP synthesis in the electron transfer chain and induce necrotic cell death . Then, the increase in lifetime of FAD autofluorescence might suggest that inner mitochondria remained functional for a prolonged period during field-induced apoptotic cell death, as reported during the drug and chemical-induced apoptosis. , The inner mitochondrial membrane remodeling and the change in ATP may have to be considered as the secondary consequences of MOMP under the nsPEF-induced apoptosis.…”

Section: Resultsmentioning

confidence: 91%

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

Awasthi

Hsu

et al. 2023

J. Phys. Chem. B

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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.

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Section: Resultsmentioning

confidence: 91%

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

Awasthi

Hsu

et al. 2023

J. Phys. Chem. B

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“…Additionally, a Correspondence to: Ippei Yagi. E-mail: yagi-3@tmu.ac.jp Department of Electrical Engineering and Computer Science, Graduate School of Systems Design, Tokyo Metropolitan University, 6-6, Asahigaoka, Hino-shi Tokyo, Japan 191-0065, an increase in intracellular Ca 2+ concentration and a decrease in Ψ m and an increase in reactive oxygen species immediately after nsPEF application have recently been reported using the technique of observing changes over time in fluorescence-stained cells [5,6].…”

Section: Introductionmentioning

confidence: 99%

“…It has also been shown that the decrease in mitochondrial membrane potential

{\Delta \varPsi}_{\mathrm{m}}

is caused by the release of

{\mathrm{Ca}}^{2+}

into the cytosol and that the frequency component in nsPEF corresponds to the shift in

{\Delta \varPsi}_{\mathrm{m}}

[1,4]. Additionally, an increase in intracellular

{\mathrm{Ca}}^{2+}

concentration and a decrease in

{\Delta \varPsi}_{\mathrm{m}}

and an increase in reactive oxygen species immediately after nsPEF application have recently been reported using the technique of observing changes over time in fluorescence‐stained cells [5,6].…”

Section: Introductionmentioning

confidence: 99%

Effects of Pulse Width and Electrical Energy of Low‐Voltage Nanosecond Pulsed Electric Fields on Mitochondria in Cancer Cells

Ninagawa,

Sugiura,

Kato

et al. 2024

IEEJ Transactions Elec Engng

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Nanosecond pulsed electric fields (nsPEFs) have recently shown promise as a cancer therapy. Although nsPEF‐induced apoptotic responses have been observed, the stress on individual cell components has not been quantified. Therefore, the authors built a cell equivalent circuit, including mitochondria, to quantitatively calculate the voltage, electric field, and electrical energy applied to the cell components in the frequency domain. Additionally, pulse width and electrical energy changed in mitochondrial membrane potential over time. The results indicated that the change of pulsed voltage switched the cell stimulation pathway, resulting in different mitochondrial membrane potentials over time. The frequency response analysis confirmed that the potential on mitochondrial membranes increased under shorter pulse width conditions. This suggested that the frequency analysis provided in this paper is useful for relating the site of cellular stimulation to induce physiological effects. © 2024 Institute of Electrical Engineer of Japan and Wiley Periodicals LLC.

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Simulation and Experimental Study on Damage of Subcellular Structure by High Voltage and Extremely Short Pulse Electric Field

Ma,

Wang,

Chen

et al. 2023

Lecture Notes in Electrical Engineering

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

Cited by 7 publications

References 51 publications

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

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

Effects of Pulse Width and Electrical Energy of Low‐Voltage Nanosecond Pulsed Electric Fields on Mitochondria in Cancer Cells

Simulation and Experimental Study on Damage of Subcellular Structure by High Voltage and Extremely Short Pulse Electric Field

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