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目的:初步探究新型物理治疗技术中频交变电流对肿瘤细胞和正常细胞体外存活率的影响;设置不同实验参数(频率、电流、时间),观察影响效果;针对实验结果,分析中频交变电流抑制肿瘤细胞体外增殖的可能机制。方法:将对数生长期的细胞悬液接种于24孔培养板,贴壁后(24 h)施加不同参数的中频电流刺激,噻唑蓝(MTT)法检测细胞存活率;以人乳腺癌细胞MCF-7为例,观察中频交变电流对细胞生存环境温度和pH的影响,Annexin V-FITC/PI双染法流式细胞术定量检测细胞凋亡/死亡情况,扫描电子显微镜和透射电子显微镜观察细胞结构变化。结果:中频交变电流能降低肿瘤细胞的体外存活率,诱导其凋亡,但不影响正常细胞(L929)存活率;MCF-7细胞体外实验最适参数是100 kHz,50 mA,每天30 min;电刺激后MCF-7细胞表面微绒毛减少,细胞线粒体肿胀,有多泡体及溶酶体形成。结论:中频交变电流能抑制肿瘤细胞增殖,但不影响正常细胞(L929);抑制效果和电刺激的频率、电流和时间有关;抑制机制可能是中频交变电流能够诱导细胞凋亡,影响细胞膜表面蛋白功能,但不影响细胞生存环境。中频交变电流抑制肿瘤体外增殖为实体瘤等增生性疾病的治疗提供了新的思路。
OBJECTIVE: To investigate the effect of the new alternating current on the viability of tumor cells and normal cells in vitro, and to set up different experimental parameters (frequency, current and time) to observe the effect. In view of the experimental results, Possible Mechanism of Tumor Cell Proliferation in. Methods: The cell suspension in logarithmic growth phase was inoculated into 24-well culture plates and stimulated by medium frequency current with different parameters 24 h after adherent. Cell survival rate was measured by MTT assay. -7 as an example to observe the effects of alternating current with medium frequency on the temperature and pH of the living environment of the cells. The apoptosis / death was quantitatively determined by flow cytometry with Annexin V-FITC / PI double staining. Scanning electron microscopy and transmission electron microscopy Cell structure changes. Results: The alternating current of medium frequency could decrease the survival rate of tumor cells in vitro and induce apoptosis, but did not affect the survival rate of normal cells (L929). The optimal parameters of MCF-7 cells in vitro were 100 kHz, 50 mA, 30 min per day After the electrical stimulation, the surface of MCF-7 cells reduced microvilli, the mitochondria were swollen, and there were more vacuoles and lysosomes. CONCLUSION: The alternating current with medium frequency can inhibit the proliferation of tumor cells but not the normal cells (L929). The inhibitory effect is related to the frequency, current and time of electrical stimulation. The inhibitory mechanism may be that medium frequency alternating current can induce apoptosis and affect cell membrane Surface protein function, but does not affect cell survival environment. Alternating frequency current can inhibit tumor proliferation in vitro and provide new ideas for the treatment of proliferative diseases such as solid tumors.