Electroporation (EP) is widely applied in electrochemotherapy to enhance drug delivery by transient membrane permeabilization. However, its impact on intercellular communication mediated by extracellular microvesicles (MVs) remains poorly understood. This study investigated the effects of reversible EP on MV release and MV-mediated biological responses in human melanoma cell lines (A375, Me45) and keratinocytes (HaCaT). MVs were isolated by differential centrifugation and characterized by flow cytometry, dynamic light scattering, transmission electron microscopy, and CD63 detection. Functional effects of EP-derived MVs on recipient cells were evaluated using viability, migration, adhesion, and real-time impedance assays. Changes in focal adhesion kinase (FAK) levels and E- and N-cadherin expression were analyzed by ELISA and flow cytometry. Reversible EP significantly increased MV release in a time- and cell line–dependent manner without altering vesicle size distribution. MVs derived from non-electroporated melanoma cells promoted proliferation, whereas EP-derived MVs inhibited proliferation, migration, and adhesion of melanoma cells. These effects were accompanied by reduced FAK levels and modulation of cadherin expression. Keratinocytes showed distinct responses, including a cadherin switch characterized by decreased E-cadherin and increased N-cadherin expression. Reversible electroporation alters not only MV secretion but also their biological activity, inducing MV-mediated bystander effects that suppress key pro-metastatic properties of melanoma cells. These findings suggest that electroporation may modulate the tumor microenvironment beyond membrane permeabilization, supporting its safety and therapeutic potential while highlighting cell type–specific effects that warrant further investigation. • Reversible electroporation increases microvesicle release in melanoma cells. • EP-derived microvesicles inhibit melanoma cell migration and proliferation. • Microvesicles mediate a bystander effect after electroporation. • Electroporation-modified microvesicles reduce focal adhesion kinase levels. • Keratinocytes exhibit cadherin switch in response to EP-derived microvesicles.
Szwedowicz et al. (Thu,) studied this question.