The effect of high frequency steep pulsed electric fields on in vitro and in vivo antitumor efficiency of ovarian cancer cell line skov3 and potential use in electrochemotherapy

Background: Patients received electrochemotherapy often associated with unpleasant sensations mainly result from low-frequency electric pulse induced muscle contractions. Increasing the repetition frequency of electric pulse can reduce unpleasant sensations. However, due to the specificity of SPEF, frequency related antitumor efficiency need to be further clarified. The aim of this study was to compare in vitro cytotoxic and in vivo antitumor effect on ovarian cancer cell line SKOV3 by SPEF with different repetition frequencies. Explore potential benefits of using high frequency SPEF in order to be exploitable in electrochemotherapy. Methods: For in vitro experiment, SKOV3 cell suspensions were exposed to SPEF with gradient increased frequencies (1, 60, 1 000, 5 000 Hz) and electric field intensity (50, 100, 150, 200, 250, 300, 350, 400 V/cm) respectively. For in vivo test, SKOV3 subcutaneous implanted tumor in BALB/c nude mice (nu/nu) were exposure to SPEF with gradient increased frequencies (1, 60, 1 000, 5 000 Hz) and fixed electric field intensity (250 V/cm) (7 mice for each frequency and 7 for control). Antitumor efficiency was performed by in vitro cytotoxic assay and in vivo tumor growth inhibition rate, supplemented by histological and TEM observations. Data were analyzed using one-way ANOVA followed by the comparisons of multiple groups. Results: SPEF with a given frequency and appropriate electric field intensity could achieve similar cytotoxicity until reached a plateau of maximum cytotoxicity (approx. 100%). SPEF with different frequencies had significant antitumor efficiency in comparison to the control group (P < 0.05). However, there was no difference in tumor responses among test groups (P > 0.05). Histological and TEM observations demonstrated obvious cell damages in response to SPEF exposure. Furthermore, SPEF with 5 kHz could induce apoptosis under TEM observations both in vitro and in vivo. Conclusion: SPEF with high frequency could also achieve similar antitumor efficiency which can be used to reduce unpleasant sensations in tumor electrical treatment. Our research proposed potential applications of using high frequency SPEF in clinical cancer treatment.