Inhibitory Effects of Sub-Nanosecond Pulsed Electric Field on Hela Cells at Elevated Temperature
Guo Fei1*, Zhang Lin1, Liu Xin1, Zhang Yu2
1 Chongqing Key Laboratory of Complex Systems and Bionic Control, Chongqing University of Posts and Telecommunications, Chongqing 400065, China; 2 Department of Gynecology and Obstetrics, Chongqing Traditional Chinese Medicine Hospital, Chongqing 400021, China
Abstract:In order to study the killing effects of combined exposure of sub-nanosecond pulsed electric field (sub-nsPEF) and temperature on HeLa cells, sub-nsPEFs (field intensity of 25, 50 and 100 kV/cm, pulse duration of 1 ns, pulse number of 4 000, 1 000 and 250, repetition frequency of 5 Hz) were applied on HeLa cells at different temperatures (25 ℃, 43 ℃, 48 ℃). The inhibition rate of cell proliferation was determined by MTT assay; morphological changes of the cells were examined by acridine orange and ethidium bromide (AO/EB) staining and transmission electron microscopy; protein expression of caspase-3 was detected by immunocytochemistry. Experimental results showed that the cell death rate was increased over time when the culture temperature reached 48℃. Next, sub-nsPEF (field intensity of 50 kV/cm, pulse duration of 1 ns, pulse number of 1 000; repetition frequency of 5 Hz) was applied on cells at different temperatures. The cell death rate was 18.07%±1.98% when the temperature reached 43℃ (P<0.05), and increased to 25.11%±6.05% when the temperature was reached 48℃ (P<0.01). The cell death rate was proportionally correlated with the field intensity when the sub-nsPEF with same energy was applied on the cells at the same temperature. The largest cell death rate of 31.09%±5.03% was obtained with the field intensity was 100 kV/cm (P<0.01). Mechanistic study indicated that the cells underwent apoptosis after the combined treatment. These data demonstrate that the sub-nsPEF with lower field intensity and temperature can cause tumor cell death through inducing apoptosis.
郭飞, 张琳, 刘欣, 张玉. 亚纳秒脉冲电场联合温度杀伤人宫颈癌细胞的实验研究[J]. 中国生物医学工程学报, 2020, 39(3): 335-341.
Guo Fei, Zhang Lin, Liu Xin, Zhang Yu. Inhibitory Effects of Sub-Nanosecond Pulsed Electric Field on Hela Cells at Elevated Temperature. Chinese Journal of Biomedical Engineering, 2020, 39(3): 335-341.
[1] 姚陈果, 赵亚军, 李成祥, 等.不可逆电穿孔微创消融肿瘤技术的研究进展 [J]. 高电压技术, 2014, 40(12): 3725-3837. [2] Aguado-Romeo MJ, Benot-López S, Romero-Tabares A. Electrochemotherapy for the treatment of unresectable locoregionally advanced cutaneous melanoma: A systematic review [J]. Actas Dermo-Sifiliográficas, 2017, 108(2): 91-97. [3] Mali B, Jarm T, Snoj M, et al. Antitumor effectiveness of electrochemotherapy: A systematic review and meta-analysis [J]. European Journal of Surgical Oncology (EJSO), 2013, 39(1): 4-16. [4] Rolong A, Rubinsky B, Davalos RV. Tissue ablation by irreversible electroporation [J]. Handbook of Electroporation, 2017, 1(1): 1-15. [5] Frühling P, Nilsson A, Duraj F, et al. Single-center nonrandomized clinical trial to assess the safety and efficacy of irreversible electroporation (IRE) ablation of liver tumors in humans: short to mid-term results [J]. European Journal of Surgical Oncology (EJSO), 2017, 43(4), 751-757. [6] Kim K, Lee WG. Electroporation for nanomedicine: A review [J]. Journal of Materials Chemistry B, 2017, 5(15): 2726-2738. [7] Dai Jie, Wu Shan, Kong Yan, et al. Nanosecond pulsed electric fields enhance the anti-tumour effects of the mTOR inhibitor everolimus against melanoma[J]. Scientific Reports, 2017, 7(1): 1-10. [8] Altunc S, Baum CE, Buchenauer CJ, et al. Design of a special dielectric lens for concentrating a subnanosecond electromagnetic pulse on a biological target [J]. IEEE Transactions on Dielectrics and Electrical Insulation, 2009, 16(5):1364-1375. [9] 郭飞, 姚陈果, 刘泽辉, 等.采用冲激脉冲辐射天线实现皮秒脉冲电场生物组织聚焦的仿真分析 [J]. 高电压技术, 2013, 39(1): 175-180. [10] 郭飞, 李成祥, 姚陈果, 等. 损耗介质透镜实现皮秒脉冲电场在乳房组织模型中的聚焦[J]. 高电压技术, 2015, 41(4): 1377-1382. [11] Schoenbach KH, Xiao S, Joshi RP, et al. The effect of intense subnanosecond electrical pulses on biological cells [J]. IEEE Transactions on Plasma Science, 2008, 36(2): 414-422. [12] Xiao Shu, Guo Siqi, Nesin V, et al. Subnanosecond electric pulses cause membrane permeabilization and cell death[J]. IEEE Transactions on Biomedical Engineering, 2011, 58(5): 1239-1245. [13] 郭飞, 姚陈果, 章锡明, 等. 高强度皮秒脉冲电场诱导 HeLa 细胞生物电效应分析[J]. 高电压技术, 2012, 38(12): 3381-3386. [14] Hua YY, Wang XS, Zhang Y, et al. Intense picosecond pulsed electric fields induce apoptosis through a mitochondrial-mediated pathway in HeLa cells [J]. Molecular Medicine Reports, 2012, 5(4): 981-987. [15] Semenov I, Xiao S, Kang D, et al. Cell stimulation and calcium mobilization by picosecond electric pulses [J]. Bioelectrochemistry, 2015, 105: 65-71. [16] Yao C, Zhao Z, Dong S, et al. High-voltage subnanosecond pulsed power source with repetitive frequency based on Marx structure [J]. IEEE Transactions on Dielectrics and Electrical Insulation, 2015, 22(4): 1896-1901. [17] 姚陈果, 赵东阳, 米彦, 等. 高压皮秒脉冲发生器的设计与实现[J]. 高电压技术, 2010, 36(7): 1698-1703. [18] 郭飞, 姚陈果, 章锡明, 等. 高强度皮秒脉冲电场诱导 HeLa 细胞生物电效应分析[J]. 高电压技术, 2012, 38(12): 3381-3386. [19] Chen WJ, Xiong ZA, Zhang M, et al. Picosecond pulsed electric fields induce apoptosis in HeLa cells via the endoplasmic reticulum stress and caspase-dependent signaling pathways[J]. International Journal of Oncology, 2013, 42(3): 963-970. [20] Jia J, Xiong ZA, Qin Q, et al. Picosecond pulsed electric fields induce apoptosis in a cervical cancer xenograft [J]. Molecular Medicine Reports, 2015, 11(3): 1623-1628. [21] Song J, Joshi RP. Temperature effects on high-intensity, ultrashort electric pulse induced cell death [C] // 2014 40th Annual Northeast Bioengineering Conference (NEBEC). Boston: IEEE, 2014: 487-488. [22] Song J, Joshi RP, Schoenbach KH. Synergistic effects of local temperature enhancements on cellular responses in the context of high-intensity, ultrashort electric pulses[J]. Medical & Biological Engineering & Computing, 2011, 49(6): 713-718. [23] Yin Shengyong, Miao Xudong, Zhang Xueming, et al. Environmental temperature affects physiology and survival of nanosecond pulsed electric field-treated cells [J]. Journal of Cellular Physiology, 2018, 233(2):1179-1190.