Numerous types of cancer confirm that every tissue in the human body has carcinogenic potential. Malignant transformation, also known as carcinogenesis, results from the failure of repair mechanisms to properly correct mutations in cells, which arise due to various factors. Therefore, efforts to overcome these deficiencies and uncover new forms of cancer treatment are ongoing. A promising treatment method, already in the clinical testing phase, is electroporation-mediated gene therapy, also known as gene electrotransfer (GET). This technique uses electrical pulses to create pores in the membranes of cancer cells, allowing the introduction of therapeutic nucleic acids (DNA, RNA, mRNA, siRNA, antisense oligonucleotides, aptamers) with the aim of inhibiting or destroying the malignancy. Electroporation and the associated transport of exogenous molecules are determined by physical factors and tissue physiology. Therefore, we were interested in how different parameters of GET electrical pulses affect the survival of tumour and normal cells, as well as the efficiency of their transfection. We determined the cytotoxicity and efficiency of individual GET protocol based on the percentage of surviving cells or transfected cells and by quantifying the fluorescence intensity of the reporter protein GFP, which we introduced into the cells in the form of previously isolated plasmid DNA. Electroporation was performed using a Genedrive electroporator with two flat parallel electrodes according to protocols with pulses of electric field strengths for different electrodes in the clinic (1000 V/cm or 1200 V/cm) and with bipolar pulses. Last pulse parameters were selected because they are characteristic of real clinical pulses for flat electrodes. All the mentioned protocols proved to be statistically significantly cytotoxic, which is a result of the transfer of exogenous DNA into cells. The highest percentage of transfection (approximately 35%) was observed in B16F10 cells. Transfection with bipolar pulses was the least effective. At the same level of transfection with the same electrical pulse parameters between cell lines, we observed significance in two lines.
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