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| 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 |
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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.
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Received: 30 July 2018
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