低强度超声联合微泡的声空化效应介导大鼠C_6胶质瘤细胞凋亡的研究  

Mediation of Acoustic Cavitation Caused by Low-intensity Ultrasound Combined with Microbubbles in Apoptosis of Rat C_6 Glioma Cells

作　　者：吴晓辉[1] 陶裕川[1] 汪峰[1] 黄琴[1] 邓金木[1] 王志刚[2] 程远[1] 

机构地区：[1]重庆医科大学附属第二医院神经外科,重庆400010 [2]重庆医科大学超声影像研究所,重庆400010

出　　处：《华中科技大学学报（医学版）》2014年第1期6-11,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30870723)

摘　　要：目的探讨不同低强度超声联合微泡的声空化效应对大鼠C6胶质瘤细胞增殖和凋亡的影响及其可能机制。方法采用不同低强度超声联合微泡辐射体外培养的大鼠C6胶质瘤细胞,四甲基偶氮唑盐(MTT)比色法检测C6胶质瘤细胞生长抑制率,吖啶橙/溴化乙啶(AO/EB)双染色法计算细胞凋亡率和坏死率,流式细胞仪分析细胞周期变化、凋亡及凋亡增殖比(APR),脱氧核糖核酸断裂的原位末端标记法(TUNEL法)观察细胞凋亡形态学改变,透射电镜及扫描电镜观察细胞超微结构变化。结果细胞生长抑制率随超声辐照强度的增强而增加;0.7 W/cm2超声辐照后细胞凋亡率最高,为(17.03±1.94)%,凋亡峰出现在G0/G1峰前且明显,细胞生长受阻于G1/S期;透射电镜观察辐照组细胞可见凋亡各阶段的典型改变;扫描电镜示辐照组细胞膜有穿孔现象。结论低强度超声辐照及联合微泡所产生的声空化效应可诱导C6胶质瘤细胞凋亡。Objective To explore the effects of the acoustic cavitation caused by low-intensity ultrasound（LIUS） combined with microbubbles on the cell growth and apoptosis of rat C6 glioma cells and the possible mechanism. Methods Rat C6glioma cells were irradiated by LIUS of different intensities. The growth inhibition rate of C6 glioma cells was detected by MTT colori- metric assay. The cell apoptosis rate and the necrosis rate were measured by AO/EB double staining,and the tumor cell cycle changes,the tumor cell apoptosis and the apoptosis/proliferation rate（APR）were determined by flow cytometry. The morpho- logical changes of glioma cells were observed by using TUNEL method and the ultrastructural changes of C6 glioma cells by transmission electron microscopy. Results Within a certain ultrasonic intensity range, the growth inhibition rate of C6 glioma cells was increased with the ultrasonic intensity elevated. The apoptosis rate（17.03± 1.94）% was highest in C6 glioma cells ir- radiated by LIUS at the intensity of 0.7 W/cm^2. An obvious apoptosis peak appeared before the Go/G1 peak. The cell growth was blocked at G1/S phase. Under transmission electron microscope,the typical changes of the cell apoptosis at various phases were found in radiation group, and the scanning electron microscopy revealed the perforation of the membrane in radiated cells. Conclusion The acoustic cavitation caused by the interaction between the LIUS and the microbubbles can induce the ap- optosis of rat C6 glioma cells.

关 键 词：低频超声 微泡 声空化效应 细胞凋亡 细胞增殖 

分 类 号：R739.41[医药卫生—肿瘤]