Faculty of Medicine, Wroclaw Medical University, 50-367 Wroclaw, Poland.
Department of Molecular and Cellular Biology, Wroclaw Medical University, 50-556 Wroclaw, Poland.
Molecules. 2020 Nov 19;25(22):5406. doi: 10.3390/molecules25225406.
Electroporation, applied as a non-thermal ablation method has proven to be effective for focal prostate treatment. In this study, we performed pre-clinical research, which aims at exploring the specific impact of this so-called calcium electroporation on prostate cancer. First, in an in-vitro study of DU 145 cell lines, microsecond electroporation (μsEP) parameters were optimized. We determined hence the voltage that provides both high permeability and viability of these prostate cancer cells. Subsequently, we compared the effect of μsEP on cells' viability with and without calcium administration. For high-voltage pulses, the cell death's mechanism was evaluated using flow-cytometry and confocal laser microscopy. For lower-voltage pulses, the influence of electroporation on prostate cancer cell mobility was studied using scratch assays. Additionally, we applied calcium-binding fluorescence dye (Fluo-8) to observe the calcium uptake dynamic with the fluorescence microscopy. Moreover, the molecular dynamics simulation visualized the process of calcium ions inflow during μsEP. According to our results calcium electroporation significantly decreases the cells viability by promoting apoptosis. Furthermore, our data shows that the application of pulsed electric fields disassembles the actin cytoskeleton and influences the prostate cancer cells' mobility.
电穿孔作为一种非热消融方法已被证明对前列腺焦点治疗有效。在这项研究中，我们进行了临床前研究，旨在探索这种所谓的钙电穿孔对前列腺癌的具体影响。首先，在 DU 145 细胞系的体外研究中，优化了微秒电穿孔 (μsEP) 参数。因此，我们确定了既能提供高通透性又能保持这些前列腺癌细胞活力的电压。随后，我们比较了钙给药与否对细胞活力的影响。对于高压脉冲，使用流式细胞术和共聚焦激光显微镜评估细胞死亡的机制。对于低压脉冲，使用划痕实验研究电穿孔对前列腺癌细胞迁移的影响。此外，我们应用钙结合荧光染料（Fluo-8）通过荧光显微镜观察钙摄取的动态。此外，分子动力学模拟可视化了 μsEP 过程中钙离子流入的过程。根据我们的结果，钙电穿孔通过促进细胞凋亡显著降低细胞活力。此外，我们的数据表明，脉冲电场的应用会破坏肌动蛋白细胞骨架并影响前列腺癌细胞的迁移能力。
