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Bystander Effect Induced by Electroporation is Possibly Mediated by Microvesicles and Dependent on Pulse Amplitude,
Ajda Prevc1, Apolonija Bedina Zavec2, Maja Cemazar1,3
1Department of Experimental Oncology, Institute of Oncology Ljubljana, Zaloska ulica 2, 1000, Ljubljana, Slovenia.
Abstract:
Bystander effect, a known phenomenon in radiation biology, where irradiated cells release signals which cause damage to nearby, unirradiated cells, has not been explored in electroporated cells yet. Therefore, our aim was to determine whether bystander effect is present in electroporated melanoma cells in vitro, by determining viability of non-electroporated cells exposed to medium from electroporated cells and by the release of microvesicles as potential indicators of the bystander effect. Here, we demonstrated that electroporation of cells induces bystander effect: Cells exposed to electric pulses mediated their damage to the non-electroporated cells, thus decreasing cell viability. We have shown that shedding microvesicles may be one of the ways used by the cells to mediate the death signals to the neighboring cells. The murine melanoma B16F1 cell line was found to be more electrosensitive and thus more prone to bystander effect than the canine melanoma CMeC-1 cell line. In B16F1 cell line, bystander effect was present above the level of electropermeabilization of the cells, with the threshold at 800 V/cm. Furthermore, with increasing electric field intensities and the number of pulses, the bystander effect also increased. In conclusion, electroporation can induce bystander effect which may be mediated by microvesicles, and depends on pulse amplitude, repetition frequency and cell type.
Insights
Electroporation induces a bystander effect in melanoma cells, where electroporated cells damage neighboring non-electroporated cells. This effect, potentially mediated by microvesicles, varies by cell type and electric pulse parameters.
Area of Science:
- Cell Biology
- Biophysics
- Cancer Research
Background:
- The bystander effect, known in radiation biology, involves irradiated cells damaging nearby unirradiated cells.
- This phenomenon has not been previously investigated in the context of electroporation.
Purpose of the Study:
- To determine if a bystander effect exists in electroporated melanoma cells in vitro.
- To investigate the role of microvesicles in mediating this effect.
Main Methods:
- Assessing the viability of non-electroporated cells exposed to medium from electroporated cells.
- Analyzing the release of microvesicles from electroporated cells.
- Comparing the electrosensitivity and bystander effect in murine melanoma B16F1 and canine melanoma CMeC-1 cell lines.
Main Results:
- Electroporation of melanoma cells induces a bystander effect, decreasing the viability of non-electroporated neighboring cells.
- Microvesicle shedding is identified as a potential mechanism for mediating damage signals.
- Murine melanoma B16F1 cells are more susceptible to electroporation-induced bystander effects than canine melanoma CMeC-1 cells.
- The bystander effect in B16F1 cells was observed above 800 V/cm and increased with higher electric field intensities and pulse numbers.
Conclusions:
- Electroporation can induce a bystander effect in melanoma cells.
- Microvesicles may mediate this effect, influencing cell viability.
- The magnitude of the bystander effect is dependent on electric pulse parameters and cell type.
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