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目的:观察极低频磁场对人肝癌细胞株BEL-7402的作用,探讨其抑制细胞生长的机制。方法:建立极低频磁场生物模型。极低频磁场处理(100Hz,0.7mT,30min)人肝癌细胞系BEL-7402后,给予不同剂量X射线照射(2、4、6、8和10Gy)。作细胞集落形成试验;流式细胞仪研究极低频磁场作用下人肝癌细胞系BEL-7402细胞周期及相关蛋白的改变;RT-PCR检测p53水平。结果:细胞集落形成试验结果显示,极低频磁场加X射线组抑制率为73.00%、78.00%、58.00%、55.00%和67.00%,单纯X射线组抑制率为38.00%、41.00%、20.00%、24.00%和30.00%,F=5.89,P=0.03。流式细胞仪检测显示,5种不同放射强度下,极低频磁场加放射组凋亡率为10.00%、14.50%、4.30%、5.10%和7.10%,单纯放射组凋亡率为0.10%、8.10%、0.10%、0.41%和2.20%,极低频磁场加X射线对肝癌细胞株BEL-7402的凋亡率均高于单纯放射,2和4Gy照射时最为明显,两组差异有统计学意义,F=6.9,P=0.03。极低频磁场组细胞生长停滞于G2/M期,细胞周期停滞相关的p53蛋白水平均显著增高。结论:100Hz,0.7mT极低频磁场能诱导人肝癌细胞BEL-7402产生凋亡。极低频磁场抑制癌细胞增殖作用的机制与G2/M期停滞有关。其中p53激活是引起细胞周期停滞重要原因。
OBJECTIVE: To observe the effect of extremely low frequency magnetic field on human hepatocellular carcinoma cell line BEL-7402 and to explore its mechanism of inhibiting cell growth. Methods: To establish a low frequency magnetic field biological model. After ultra-low frequency magnetic field treatment (100 Hz, 0.7 mT, 30 min), the human hepatoma cell line BEL-7402 was irradiated with different doses of X rays (2, 4, 6, 8 and 10 Gy). Cell colony formation assay was performed. Flow Cytometry was used to study the cell cycle and related proteins in human hepatocellular carcinoma cell line BEL-7402 under the effect of ultra-low frequency magnetic field. The level of p53 was detected by RT-PCR. Results: The results of colony formation assay showed that the inhibition rates of ultra-low frequency magnetic field and X-ray group were 73.00%, 78.00%, 58.00%, 55.00% and 67.00% respectively. The inhibitory rates were 38.00%, 41.00% and 20.00% 24.00% and 30.00%, F = 5.89, P = 0.03. The results of flow cytometry showed that the apoptotic rates in the ultra-low frequency magnetic field plus radiation group were 10.00%, 14.50%, 4.30%, 5.10% and 7.10% at 5 different radiances, and those in the radiotherapy group were 0.10% and 8.10 %, 0.10%, 0.41% and 2.20% respectively. The rate of apoptosis in BEL-7402 cells treated by X-ray and extra-low frequency magnetic field was higher than that of pure radiotherapy alone. The most obvious difference was observed between 2 and 4 Gy irradiation. F = 6.9, P = 0.03. In low frequency magnetic field group, the cell growth arrested in G2 / M phase, and the cell cycle arrest related p53 protein levels were significantly increased. Conclusion: A 100 Hz, 0.7 mT extra-low frequency magnetic field can induce apoptosis in human hepatoma BEL-7402 cells. The mechanism of extremely low frequency magnetic field inhibiting the proliferation of cancer cells is related to G2 / M arrest. Among them, p53 activation is an important cause of cell cycle arrest.