摘要
目的:采用原子力声显微镜及透射电镜观察低频低能量超声联合微泡对前列腺癌细胞DU145及正常前列腺上皮细胞RWPE-1的作用。方法:两种细胞均分为对照组、单纯超声组、超声联合微泡组。对照组加入一定比例的生理盐水,不进行超声辐照;单纯超声组加入相同比例的生理盐水,用发射频率为21 k Hz的低频超声辐照,辐照2 min,占空比30%;超声联合微泡组加入同前相同比例的微泡造影剂悬浊液,用与单纯超声组相同的超声辐照。处理过的细胞立即用原子力声显微镜观察其形貌并计算其杨氏模量。同时,对同一处理方式的对照组及超声联合微泡组细胞继续培养24 h后,用透射电镜观察细胞。结果:单纯超声组的DU145细胞及RWPE-1细胞的细胞形态与对照组无明显变化;超声联合微泡组的DU145细胞及RWPE-1细胞形态呈类圆形,细胞表面可见放射状显微丝状结构,细胞膜表面可见多个大孔状结构;超声联合微泡组的DU145细胞的弹性模量较RWPE-1细胞大,且与对照组相比,两种细胞的弹性模量均变大,DU145细胞尤甚。透射电镜观察结果示对照组的两种细胞未见细胞自噬,而超声联合微泡组两种细胞都出现自噬现象,其中DU145细胞中可见凋亡现象。结论:低频低能量超声联合微泡可引起前列腺癌DU145细胞及正常前列腺上皮RWPE-1细胞的细胞膜出现孔状结构,并诱导其自噬,且对DU145细胞的损伤大于RWPE-1细胞。
Objective： To explore the impact of 21 kHz low-intensity ultrasound combined with microbubbles on both DU145 cells and RWPE-1 cells by using Atomic force acoustic microscope （AFAM） and Transmission electron microscopy （TEM）.Methods： Both DU145 cells and RWPE-1 cells were divided into three groups： control group, ultrasound group and ultrasound combined with microbubbles group. Conditions of the low-frequency and low-energy ultrasound （US） irradia-tion were： Frequency, 21 kHz; Exposure time, 2 min at a duty ratio of 30%; and the valid treatment time of 84 seconds was used for the combination with microbubbles （100μl/ml）. Cells of different groups were imaged in cover slips by AFAM immediately after the intervention. Twenty-four hours after intervention, TEM was used to observe cells.Results： Cellular membrane and three-dimensional structure of both cells showed significant difference in ultrasound combined with mi-crobubbles group compared with control group, while no difference in ultrasound group compared with control group. Compared with control group, the elasticity modulus of both cells in ultrasound combined with microbubbles group was higher, and also the elasticity modulus of DU145 cells was higher than RWPE-1 in ultrasound combined with microbubbles group. Lots of autophagosomes or autolysosomes were detected by TEM in both cells in ultrasound combined with mi-crobubbles group. TEM also demonstrated some apoptosis of DU145 cells in ultrasound combined with microbubbles group.Conclusions： The low-frequency ultrasound combined with microbubbles can induce autophagy in both DU145 cells and RWPE-1 cells, and the injury to DU145 cells is more serious than to RWPE-1 cells.
出处
《声学技术》
CSCD
北大核心
2015年第4期333-337,共5页
Technical Acoustics
基金
国家自然科学基金项目(81401421)
教育部项目(20120073120100)
上海市科委自然基金面上项目(12ZR1422600)资助
关键词
低频低能量超声
微泡造影剂
前列腺癌
原子力声显微镜
细胞自噬
low-frequency ultrasound
microbubble contrast agent
prostate cancer
Atomic Force Acoustic Microscope（AFAM）
autophage
