高强度聚焦超声破坏结肠癌细胞株的实验观察

Observation of colon cancer cell line irradiated with high intensity focused ultrasound

目的:探讨用高强度聚焦超声辐照肿瘤细胞,制备肿瘤抗原的合适剂量。方法:实验于2003-12/2004-10在泰安市中心医院中心实验室和超声治疗中心完成。实验仪器为FEP-BY01型高强度超声聚焦热疗肿瘤治疗机。CT-26肿瘤细胞为BALB/C小鼠来源的结肠癌细胞,高强度聚焦超声辐照CT-26结肠癌细胞株后,用椎虫蓝染色镜下观察不同辐照时间、不同声强辐照后细胞的拒染率和形态学的变化、MTT法测定肿瘤细胞存活率,分析高强度聚焦超声剂量与肿瘤细胞存活率的关系。结果:①高强度聚焦超声声强与细胞存活率的关系:进行回归曲线拟合分析为指数关系,方程为y=155.4le-0.048,R2=0.9728,P<0.05。②高强度聚焦超声辐照时间与细胞存活率的关系:进行回归分析为直线关系,方程为y=73.45-2.462x,R2=0.918,P<0.01。随着高强度聚焦超声辐照剂量的增加肿瘤细胞的存活率迅速减少,肿瘤细胞的碎片逐渐增多。超声剂量为1000W/cm2×30s时,无细胞存活,肿瘤细胞完全失去细胞形态,全部被撕裂成碎片。结论:高强度聚焦超声能灭活肿瘤细胞且能破碎肿瘤细胞,高强度聚焦超声制备肿瘤抗原的超声剂量为1000W/cm2×30s。

AIM： To discuss the appropriate dosage of high intensity focused ultrasound （HIFU） irradiating on tumor cells in making tumor antigen. METHODS： The experiment was conducted in Central Laboratory and Ultrasound Treatment Center, Taian Municipal Central Hospital from December 2003 to October 2004. The machine was FEP-BYO1 high-energy focused ultrasound on tumor treatment machine. CT-26 tumor cell was the colon cancer cell, which originated from BALB/C mice. After irradiating cell line CT26 with HIFU, the changes of resists dyeing rate and morphology of cells observed after various irradiating time and various irradiating sound intensity under typan blue staining. Number of survival cells was detected with MTT colorimetry. Relationship between HIFU dosage and survival rate of tumor cell was analyzed. RESULTS： ①Relationship between HIFU sound intensity and survival rate of tumor cell： Regression curve fitting analysis was carried out as the index relation. The equation was y=155.4 le^-0.048, R^2=0.972 8, P〈0.05. ② Relationship between HIFU irradiation time and survival rate of tumor cell： Regression analysis was carried out as the straight line. The equation was y=73.45-2.462x, R2=0.918, P〈0.01. With increasing dose of sound intensity, cell survival rate decreased sharply and more cell debris was observed. In 1 000 W/cm^2×30 s no cell was survival, and the tumor cell totally lost cell＇s Shape, all broken into chip. CONCLUSION： The HIFU is capable of destroying and fracturing living tumor cells at one time. It was 1 000 W/cm^2×30 s that the dose of HIFU in making tumor antigen.