毫米波电磁场辐照可诱导人黑素瘤细胞凋亡
|
摘要
目的研究毫米波电磁场辐照对人黑素瘤A375细胞凋亡的影响。方法研究共分为两部分:电磁场计算实验和细胞实验。电磁场计算实验通过模拟毫米波辐照细胞这一生物物理过程,计算并对比分析比吸收率(SAR)的均匀性及强度,得出最佳辐照参数。细胞实验中,A375细胞以35.2 GHz毫米波分别辐照15、30、60、90 min,非辐照组除不接受辐照外其他环境与辐照组保持一致,caspase-3抑制剂(AC-DEVD-fmk)组在毫米波辐照90 min前以10μmoL的AC-DEVD-fmk预处理1 h。用CCK-8法检测辐照组,非辐照组及AC-DEVD-fmk预处理组的细胞活性。用Annexin-V/PI双染法检测细胞凋亡情况,蛋白印迹法检测细胞内凋亡蛋白caspase-3的表达水平。结果电磁计算得出的最佳辐照方式为沿天线线极化方向,入射场垂直于塑料培养皿底部,自下而上辐照。CCK-8细胞活性检测法的结果表明,毫米波对A375细胞活性具有抑制作用(P<0.05),且呈时间依赖性。与非辐照组相比,15、30、60、90 min毫米波辐照组A375细胞的凋亡率显著升高(P<0.05)。辐照组A375细胞中caspase-3表达水平明显增加,应用caspase-3抑制剂后可以逆转毫米波辐照对细胞活性的抑制作用,细胞活性率由(36.7±0.09)%增加到(59.8±0.06)%,与对照组相比差异具有统计学意义(P<0.05)。结论 35.2 GHz毫米波辐照可以通过激活caspase-3诱导A375细胞凋亡。
Objective To investigate the effects of millimeter wave(MMW) exposure on apoptosis of human melanoma A375 cells and explore the mechanisms. Methods Through electromagnetic field calculation we simulated MMW exposure in cells and calculated the specific absorption rate(SAR). The optimal irradiation parameters were determined according to the uniformity and intensity of the SAR. A375 cells were then exposed to MMV for 15, 30, 60, or 90 min, with or without pretreatment with the caspase-3 inhibitor AC-DEVD-fmk(10 μmol/L) for 1 h at 90 min before the exposure. CCK-8 assay was used to assess the changes in the viability and Annexin-V/PI staining was used to detect the apoptosis of the cells following the exposures; Western blotting was used to detect the expression of caspase-3 in the cells. Results The results of electromagnetic field calculation showed that for optimal MMV exposure, the incident field needed to be perpendicular to the bottom of the plastic Petri dish with the antenna placed below the dish. CCk-8 assay showed that MMW exposure significantly inhibited the cell viability in a time-dependent manner(P<0.05); exposures for 15, 30, 60, and 90 min all resulted in significantly increased apoptosis of the cells(P<0.05). The cells with MMW exposure showed significantly increased expression of caspase-3. The inhibitory effect of MMW on the cell viability was antagonized significantly by pretreatment of the cells with AC-DEVD-fmk(P<0.05), which increased the cell viability rate from(36.7 ± 0.09)% to(59.8 ± 0.06)%(P<0.05). Conclusion 35.2GHz millimeter wave irradiation induces apoptosis in A375 cells by activating the caspase-3 protein.
Objective To investigate the effects of millimeter wave(MMW) exposure on apoptosis of human melanoma A375 cells and explore the mechanisms. Methods Through electromagnetic field calculation we simulated MMW exposure in cells and calculated the specific absorption rate(SAR). The optimal irradiation parameters were determined according to the uniformity and intensity of the SAR. A375 cells were then exposed to MMV for 15, 30, 60, or 90 min, with or without pretreatment with the caspase-3 inhibitor AC-DEVD-fmk(10 μmol/L) for 1 h at 90 min before the exposure. CCK-8 assay was used to assess the changes in the viability and Annexin-V/PI staining was used to detect the apoptosis of the cells following the exposures; Western blotting was used to detect the expression of caspase-3 in the cells. Results The results of electromagnetic field calculation showed that for optimal MMV exposure, the incident field needed to be perpendicular to the bottom of the plastic Petri dish with the antenna placed below the dish. CCk-8 assay showed that MMW exposure significantly inhibited the cell viability in a time-dependent manner(P<0.05); exposures for 15, 30, 60, and 90 min all resulted in significantly increased apoptosis of the cells(P<0.05). The cells with MMW exposure showed significantly increased expression of caspase-3. The inhibitory effect of MMW on the cell viability was antagonized significantly by pretreatment of the cells with AC-DEVD-fmk(P<0.05), which increased the cell viability rate from(36.7 ± 0.09)% to(59.8 ± 0.06)%(P<0.05). Conclusion 35.2GHz millimeter wave irradiation induces apoptosis in A375 cells by activating the caspase-3 protein.
引文
[1]李璟蓉,赵瑞,方锐华,等.miR-122-5p通过靶向NOP14抑制黑素瘤的细胞增殖[J].南方医科大学学报,2018,38(11):1360-5.
[2]Mashima E,Inoue A,Sakuragi Y,et al.Nivolumab in the treatment of malignant melanoma:review of the literature[J].Onco Targets Ther,2015,8(1):2045-51.
[3]Banik S,Bandyopadhyay S,Ganguly S.Bioeffects of microwave-a brief review[J].Bioresour Technol,2003,87(2):155-9.
[4]Pakhomov AG,Akyel Y,Pakhomova ON,et al.Current state and implications of research on biological effects of millimeter waves:a review of the literature[J].Bioelectromagnetics,1998,19(2):393-413.
[5]El Khoueiry C,Moretti D,Renom R,et al.Decreased spontaneous electrical activity in neuronal networks exposed to radiofrequency1 800 MHz signals[J].J Neurophysiol,2018,120(6):2719-29.
[6]吴瑛,赖声礼.毫米波的生物效应及毫米波疗法研究进展[J].第一军医大学学报,2001,21(1):62-4.
[7]Beneduci A,Chidichimo G,de Rose R,et al.Frequency and irradiation time-dependant antiproliferative effect of low-power millimeter waves on RPMI 7932 human melanoma cell line[J].Anticancer Res,2005,25(2A):1023-8.
[8]Beneduci A,Chidichimo G,Tripepi S,et al.Antiproliferative effect of millimeter radiation on human erythromyeloid leukemia cell line K562 in culture:Ultrastructural-and metabolic-induced changes[J].Bioelectrochemistry,2007,70(2):214-20.
[9]王艳萍,袁淑兰,陈晓禾,等.毫米波辐射对鼻咽癌细胞增殖和凋亡的影响[J].四川大学学报:自然科学版,2000,35(S1):37.
[10]王艳萍,袁淑兰,陈晓禾,等.毫米波辐射对人急性早幼粒白血病细胞诱导凋亡的作用[J].中华理疗杂志,2001,24(6):21-4.
[11]方昆阳,赖声礼,罗绍凯,等.毫米波辐射对K562细胞作用机制研究[J].华南理工大学学报:自然科学版,2001,29(12):6-9.
[12]Gapeyev AB,Kulagina TP,Aripovsky AV.Exposure of tumorbearing mice to extremely high-frequency electromagnetic radiation modifies the composition of fatty acids in thymocytes and tumor tissue[J].Int J Radiat Biol,2013,89(8):602-10.
[13]Li X,Ye H,Cai L,et al.Millimeter wave radiation induces apoptosis via affecting the ratio of Bax/Bcl-2 in SW1353 human chondrosarcoma cells[J].Oncol Rep,2012,27(3):664-72.
[14]赵建勋,鲁德强.细胞培皿构形对毫米波辐照剂量的影响[J].西安电子科技大学学报:自然科学版,2004,31(1):115-8.
[15]李缉熙,牛中奇.生物电磁学概论[M].西安:西安电子科技大学出版社,1991.
[16]Espinosa EI,Linares YR.Analysis of SAR distribution in a heterogeneous and homogenous headmodel at 2.4 GHz using the FDTDmethod[J].Microw Optical Techn Letters,2018,60(6):1323-31.
[17]Angulo LD,Garcia SG,Pantoja MF,et al.Improving the sar distribution in petri-dish cell cultures[J].J Electro Wav Applica,2010,24(5/6):815-26.
[18]Zhao JX.Numerical dosimetry for cells under millimetre-wave irradiation using Petri dish exposure set-ups[J].Phys Med Biol,2005,50(14):3405-21.
[19]Hasgall PA,Di Gennaro F,Baumgartner C,et al.IT'is database for thermal and electromagnetic parameters of biological tissues[J].JCell Biol,2012,10(6):376.
[20]Zhadobov M,Augustine R,Sauleau R,et al.Complex permittivity of representative biological solutions in the 2-67GHz range[J].Bioelectromagnetics,2012,33(4):346-55.
[21]高源慈,孙嘉鸿,余国芬.细胞培养皿的介电常数测定[J].电子科技大学学报,2007,36(1):70-1,111.
[22]张明子,王晓军,管恩玲,等.不同瘢痕类型中Caspase-3,Caspase-8及PCNA的表达差异分析[J].中国美容整形外科杂志,2018,29(2):85-8.
[23]Beneduci A,Chidichimo G,Tripepi S.Antiproliferative effect of millimeter radiation on human erythromyeloid leukemia cell line K562 in culture:ultrastructural-and metabolic-induced changes[J].Bioelectrochemistry,2007,70(2):214-20.
[24]Ouyang L,Shi Z,Zhao S,et al.Programmed cell death pathways in cancer:a review of apoptosis,autophagy and programmed necrosis[J].Cell Prolif,2012,45(6):487-98.
[25]Galluzzi L,Vitale I,Aaronson SA,et al.Molecular mechanisms of cell death:recommendations of the momenclature committee on Cell Death 2018[J].Cell Death Differ,2018,25(3):486-541.
[26]Hengartner MO.The biochemistry of apoptosis[J].Nature,2000,407(6805):770-6.
[27]Julien O,Wells JA.Caspases and their substrates[J].Cell Death Differ,2017,24(8,SI):1380-9.
[28]Li J,Yuan J.Caspases in apoptosis and beyond[J].Oncogene,2008,27(48):6194-206.
[29]Li Z,Sheng M.Caspases in synaptic plasticity[J].Mol Brain,2012,5(1):15-21.
[30]Belyaev I.Non-thermal biological effects of microwaves[J].Microwave Review,2005,11(2):375-403.
[31]赵建勋,牛中奇,鲁德强.辐照到培皿中细胞单层上的毫米波功率流密度分析[J].生物医学工程学杂志,2004,21(1):97-101.
[2]Mashima E,Inoue A,Sakuragi Y,et al.Nivolumab in the treatment of malignant melanoma:review of the literature[J].Onco Targets Ther,2015,8(1):2045-51.
[3]Banik S,Bandyopadhyay S,Ganguly S.Bioeffects of microwave-a brief review[J].Bioresour Technol,2003,87(2):155-9.
[4]Pakhomov AG,Akyel Y,Pakhomova ON,et al.Current state and implications of research on biological effects of millimeter waves:a review of the literature[J].Bioelectromagnetics,1998,19(2):393-413.
[5]El Khoueiry C,Moretti D,Renom R,et al.Decreased spontaneous electrical activity in neuronal networks exposed to radiofrequency1 800 MHz signals[J].J Neurophysiol,2018,120(6):2719-29.
[6]吴瑛,赖声礼.毫米波的生物效应及毫米波疗法研究进展[J].第一军医大学学报,2001,21(1):62-4.
[7]Beneduci A,Chidichimo G,de Rose R,et al.Frequency and irradiation time-dependant antiproliferative effect of low-power millimeter waves on RPMI 7932 human melanoma cell line[J].Anticancer Res,2005,25(2A):1023-8.
[8]Beneduci A,Chidichimo G,Tripepi S,et al.Antiproliferative effect of millimeter radiation on human erythromyeloid leukemia cell line K562 in culture:Ultrastructural-and metabolic-induced changes[J].Bioelectrochemistry,2007,70(2):214-20.
[9]王艳萍,袁淑兰,陈晓禾,等.毫米波辐射对鼻咽癌细胞增殖和凋亡的影响[J].四川大学学报:自然科学版,2000,35(S1):37.
[10]王艳萍,袁淑兰,陈晓禾,等.毫米波辐射对人急性早幼粒白血病细胞诱导凋亡的作用[J].中华理疗杂志,2001,24(6):21-4.
[11]方昆阳,赖声礼,罗绍凯,等.毫米波辐射对K562细胞作用机制研究[J].华南理工大学学报:自然科学版,2001,29(12):6-9.
[12]Gapeyev AB,Kulagina TP,Aripovsky AV.Exposure of tumorbearing mice to extremely high-frequency electromagnetic radiation modifies the composition of fatty acids in thymocytes and tumor tissue[J].Int J Radiat Biol,2013,89(8):602-10.
[13]Li X,Ye H,Cai L,et al.Millimeter wave radiation induces apoptosis via affecting the ratio of Bax/Bcl-2 in SW1353 human chondrosarcoma cells[J].Oncol Rep,2012,27(3):664-72.
[14]赵建勋,鲁德强.细胞培皿构形对毫米波辐照剂量的影响[J].西安电子科技大学学报:自然科学版,2004,31(1):115-8.
[15]李缉熙,牛中奇.生物电磁学概论[M].西安:西安电子科技大学出版社,1991.
[16]Espinosa EI,Linares YR.Analysis of SAR distribution in a heterogeneous and homogenous headmodel at 2.4 GHz using the FDTDmethod[J].Microw Optical Techn Letters,2018,60(6):1323-31.
[17]Angulo LD,Garcia SG,Pantoja MF,et al.Improving the sar distribution in petri-dish cell cultures[J].J Electro Wav Applica,2010,24(5/6):815-26.
[18]Zhao JX.Numerical dosimetry for cells under millimetre-wave irradiation using Petri dish exposure set-ups[J].Phys Med Biol,2005,50(14):3405-21.
[19]Hasgall PA,Di Gennaro F,Baumgartner C,et al.IT'is database for thermal and electromagnetic parameters of biological tissues[J].JCell Biol,2012,10(6):376.
[20]Zhadobov M,Augustine R,Sauleau R,et al.Complex permittivity of representative biological solutions in the 2-67GHz range[J].Bioelectromagnetics,2012,33(4):346-55.
[21]高源慈,孙嘉鸿,余国芬.细胞培养皿的介电常数测定[J].电子科技大学学报,2007,36(1):70-1,111.
[22]张明子,王晓军,管恩玲,等.不同瘢痕类型中Caspase-3,Caspase-8及PCNA的表达差异分析[J].中国美容整形外科杂志,2018,29(2):85-8.
[23]Beneduci A,Chidichimo G,Tripepi S.Antiproliferative effect of millimeter radiation on human erythromyeloid leukemia cell line K562 in culture:ultrastructural-and metabolic-induced changes[J].Bioelectrochemistry,2007,70(2):214-20.
[24]Ouyang L,Shi Z,Zhao S,et al.Programmed cell death pathways in cancer:a review of apoptosis,autophagy and programmed necrosis[J].Cell Prolif,2012,45(6):487-98.
[25]Galluzzi L,Vitale I,Aaronson SA,et al.Molecular mechanisms of cell death:recommendations of the momenclature committee on Cell Death 2018[J].Cell Death Differ,2018,25(3):486-541.
[26]Hengartner MO.The biochemistry of apoptosis[J].Nature,2000,407(6805):770-6.
[27]Julien O,Wells JA.Caspases and their substrates[J].Cell Death Differ,2017,24(8,SI):1380-9.
[28]Li J,Yuan J.Caspases in apoptosis and beyond[J].Oncogene,2008,27(48):6194-206.
[29]Li Z,Sheng M.Caspases in synaptic plasticity[J].Mol Brain,2012,5(1):15-21.
[30]Belyaev I.Non-thermal biological effects of microwaves[J].Microwave Review,2005,11(2):375-403.
[31]赵建勋,牛中奇,鲁德强.辐照到培皿中细胞单层上的毫米波功率流密度分析[J].生物医学工程学杂志,2004,21(1):97-101.