热籽感应加热对恶性黑色素瘤细胞的杀伤作用

Killing Effect of Thermoseeds Inductive Heating on Malignant Melanoma Cells

目的探讨热籽感应加热对体外培养的恶性黑色素瘤细胞的杀伤效果。方法将体外培养的B16恶性黑色素瘤细胞分为对照组、单独磁场组、单独热籽组(一颗热籽、二颗热籽)、热籽加温组(一颗热籽、二颗热籽)6组,其中对照组、热籽加温组又分为6个时间亚组(5、10、15、20、25、30min),各组分别处理后,倒置显微镜进行形态学观察,四甲基噻唑兰比色法测定细胞活性,得出每组处理后的细胞存活率。结果实验中所用的铁磁热籽在现有参数交变磁场中具有良好的升温效果,一颗热籽培养皿内加热温度可达(53.1±0.5)℃,二颗热籽则可达(56.5±0.5)℃。热籽感应加温对体外培养的B16细胞具有明显的杀伤效应,镜下观察出现明显的凋亡或坏死的形态学改变。二颗热籽加温组20min细胞存活率为(3.1±2.9)%,25min和30min便可将培养皿内肿瘤细胞全部杀死。加热30min时,单独磁场和单独热籽对B16细胞存活率影响不大,存活率分别为(98.1±5.1)%、(99.5±2.3)%、(94.6±11.0)%,与对照组相比均无统计学差异(分别为q对照-单独磁场=0.497,P>0.05;q对照-一颗热籽=0.120,P>0.05;q对照-二颗热籽=1.419,P>0.05;q单独磁场-一颗热籽=0.377,P>0.05;q单独磁场-二颗热籽=0.922,P>0.05;q一颗热籽-二颗热籽=1.299,P>0.05);加热30min时,热籽加温组和对照组、单独磁场组、单独热籽组存活率相比有统计学差异(q对照-一颗热籽加温=16.934,P<0.01;q单独磁场-一颗热籽加温=16.437,P<0.01;q一颗热籽-一颗热籽加温=16.814,P<0.01;q二颗热籽-一颗热籽加温=15.515,P<0.01)。结论热籽感应加温交变磁场中可升温到适宜的温度,有效杀伤体外培养的黑色素瘤细胞,导致肿瘤细胞凋亡或死亡。

Objective To study the killing effect of thermoseeds inductive heating on malignant melanoma cells cultured in vitro. Methods In vitro cultured B16 malignant melanoma cells were divided into 6 groups： control, magnetic, thermoseods （one thermoseod and two thermoseeds）, and thermoseods heating （one thermoseed and two thermoseeds） groups. The control and thermoseods heating groups were further divided into six subgroups （5, 10, 15,20, 25, and 30 minutes）. Cell morphological changes were observed by inverted microscopy. The cell activity of each group was measured by MTT method after treatment. The survival rate of the cells was calculated. Results The thermoseeds used in our experiment showed a good heating ability in alternating magnetic field. Temperature of culture dishes with one and two thermoseods could reach （53.5 ± 0.5 ） ℃ and （56.5 ± 0.5 ） ℃ , respectively. The B16 melanoma cells were apparently killed or damaged by thermoseed inductive heating. Inverted microscopy showed apoptosis and necrosis of the tumor cells. The survival rate of the cells heated with two thermoseeds for 20 minutes was （ 3.1 ± 2.9 ） %. After being heated for 25 or 30 minutes, all the tumor cells cultured in the dish were killed. In the magnetic and thermoseed groups, the survival rate of the tumor cells were （98.1 ± 5.1 ） % , （99.5 ± 2.3 ） % , and （94.6 ± 11.0） % , respectively, which are not significantly lower than that in the control group （qcotrol-Magnetic = 0. 497, P 〉 0.05 ; qcontrol-One thermoaced = 0. 120, P 〉 0.05 ; qControl-Two thermosceda = 1. 419, P 〉 0.05 ; qMagnetic-One thermoseed = 0. 377, P 〉 0.05 ; qMagnetie-Two thermoseeds = 0. 922, P 〉 0.05 ; and qOne thermoseed-Two thermoseeds = 1. 299, P 〉 0.05 ）. After being heated for 30 minutes, the survival rate of the thermoseods heating group was significantly lower than that in the control, magnetic, and thermoseeds groups （qControl-One thermossed heating = 16. 934, P 〈 0. 01; qMagnetic-One thermoseed heating = 16. 437, P 〈 0. 01; qOne thermoseed-One thermoseed heating =16.814, P〈0.01; and qTwo thermosseds-One thermoseedheating=15.515, P〈0.01）. Conclusions Thermoseeds inductive heating technology can achieve an appropriate temperature to effectively kill in vitro cultured melanoma cells, leading to apoptosis or death of the tumor cells.