摘要
目的:观察高强度聚焦超声对体外培养的CT-26肿瘤细胞的急性生物学效应,分析高强度聚焦超声破坏肿瘤细胞的剂量-效应关系。方法:实验于2005-05/08在泰安市中心医院中心实验室和肿瘤内科完成。CT-26肿瘤细胞为BALB/c小鼠来源的结肠癌细胞,由解放军第二军医大学免疫学研究所惠赠。固定声强(均为1000W/cm2),应用不同辐照时间(分别为1,5,10,15,20,25,30,35s)的高强度聚焦超声辐照CT-26细胞系后,用锥虫蓝染色、四甲基偶氮唑盐法测定活肿瘤细胞数,同时用锥虫蓝染色观察肿瘤细胞的形态学变化,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锥虫蓝染色后光镜下见辐照5s时有少部分细胞死亡,随着辐照时间的延长,死亡细胞逐渐增加;当辐照到20s时细胞几乎全部死亡,可见少量细胞碎片,但仍有轮廓清晰的死亡细胞;25s时细胞全部死亡,细胞碎片多见;当辐照30s及35s时肿瘤细胞全部碎裂成片,未见正常细胞形态(细胞死亡率:辐照5s时32.7%,辐照20s时93.7%,辐照25s时98.5%,辐照30和35s时100%)。②四甲基偶氮唑盐实验中根据吸光度获得细胞存活率,细胞存活率与辐照时间二者关系经SPSS10.0统计软件进行回归分析为y=70.180-2.416x(R2=0.874,P<0.01),方程有统计学意义。结论:当高强度聚焦超声声强固定为1000W/cm2时,细胞存活率随高强度聚焦超声辐照时间延长呈直线下降,1000W/cm2×30s是能灭活肿瘤细胞且使之完全破碎的剂量。
AIM： To explore the acute biological effects with different irradiation times of high intensity focused ultrasound （HIFU） on CT-26 tumor cells and investigate the relationship between the ultrasonic dosage and therapeutic effects.
METHODS： The experiment was accomplished in the Central Laboratory and Department of Medical Oncology, Taian Central Hospital from May to August in 2005. CT-26 cells of colon carcinoma in BALB/c mice were offered by the Immunology Institute of the Second Military Medical University of Chinese PLA. CT-26 cell line received regular sound intensity （1 000 W/cm^2） and HIFU irradiation of different irradiation times （1, 5, 10, 15, 20, 25, 30 and 35 seconds）. The number of survival tumor cells was detected with trypan blue staining and MTT colorimetry. The relations of HIFU dosage and tumor cell survival rates, as well as the morphologic change of the cells were also analyzed.
RESULTS：（1）After 5-second irradiation, few cells began to present apoptosis under the microscope. With the prolongation of irradiation time, cell survival rate decreased sharply; At 20 seconds, the cells almost died, except a small quantity of cell debris and sharp cellular edge; After 25 seconds, all the cell showed apoptosis in debris, and were broken into debris by 1 000 W/cm^2×30 seconds in irregular morphology （Cell death rate： 32.7%, 93.7%, 98.5%, 100%）.（2）The relationship between irradiation time and cell survival rate was linear in regression analysis by SPSS 10.0 software, with the significant differences （y =70.180-2.416x, R^2 =0.874, P 〈 0.01）.
CONCLUSION： With the increase of HIFU irritation time, the survival rate of tumor cells presents the linear decrease tendency. And 1 000 W/cm^2×30 seconds is the minimum dose of bringing cells into complete dead and breaking them into debris.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第9期1704-1706,共3页
Journal of Clinical Rehabilitative Tissue Engineering Research
