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近20年的动物实验及临床前期研究表明,纳秒脉冲具有能量高、微创治疗、不需联合化疗药物等优点。因此本研究将纳秒脉冲作用于MDA–MB–231乳腺癌荷瘤小鼠,观察局部原发肿瘤的消融效果,随访局部消融对免疫细胞亚群的影响情况,探索其非热消融的免疫机制。本实验在小鼠皮下接种小鼠乳腺癌MDA–MB–231细胞,2周肿瘤直径达到10 mm时进行纳秒脉冲局部消融治疗,治疗参数选择脉冲宽度为300 ns,电场强度为40 k V/cm,脉冲个数为100,治疗频率为隔日重复共3次。研究结果表明原位接种的MDA–MB–231乳腺癌细胞成瘤率达到100%。纳秒脉冲治疗后随访肿瘤大小、形态及体积至1个月,肿瘤直径达到20 mm时结束动物实验,B超监测数据显示,纳秒脉冲局部消融对原位肿瘤的消融率为83%,显著降低了原发肿瘤的体积,与对照组相比具有显著性降低;用流式细胞术检测脾脏中各淋巴细胞亚群的变化。射频消融后,小鼠脾脏CD4+T细胞、CD8+T细胞数量和比例升高,提示局部使用的纳秒脉冲刺激脾脏T细胞增殖能力。纳秒脉冲消融不但能够抑制原发肿瘤,还能增强小鼠免疫功能,使得肿瘤生长和转移侵袭力下降,有助于抑制肿瘤的生成及发展。
Nearly 20 years of animal experiments and preclinical studies have shown that nanosecond pulses with high energy, minimally invasive treatment, without the need for chemotherapy and other advantages. Therefore, in this study, nanosecond pulses were applied to MDA-MB-231 breast cancer-bearing mice to observe the ablation effect of local primary tumors and follow-up local ablation on immune cell subsets and to explore the immune mechanism of non-thermal ablation . In this study, mouse breast cancer MDA-MB-231 cells were inoculated subcutaneously in mice and local nanosecond pulse ablation was performed at 2-week tumor diameter of 10 mm. The selected pulse width was 300 ns and the electric field intensity was 40 kV / cm, the number of pulses is 100, the treatment frequency is repeated every other day for 3 times. The results show that in situ inoculation of MDA-MB-231 breast cancer cell tumorigenicity rate of 100%. After the nanosecond pulse treatment, the size, shape and volume of the tumor were followed up to 1 month, and the animal experiment was ended when the tumor diameter reached 20 mm. The B-monitoring data showed that the ablation rate of the local nanosecond pulse on the tumor in situ was 83% Reduce the volume of the primary tumor, compared with the control group was significantly reduced; using flow cytometry to detect changes in the spleen lymphocyte subsets. After radiofrequency ablation, the number and proportion of CD4 + T cells and CD8 + T cells in spleen of mice increased, suggesting that local use of nanosecond pulses could stimulate the proliferation of splenic T cells. Nanosecond pulse ablation not only can inhibit the primary tumor, but also enhance the immune function of mice, making tumor growth and metastasis invasiveness decreased, helps to inhibit tumor formation and development.