低强度超声增加前列腺癌细胞膜通透性的体外实验研究  

Permeability increase of prostate cancer cells membrane under low intensity ultrasound in vitro

作　　者：张吉臻[1] 申锷[1] 胡兵[1] 

机构地区：[1]上海交通大学附属第六人民医院超声科,上海超声医学研究所,上海200233

出　　处：《中华医学超声杂志（电子版）》2010年第12期15-18,共4页Chinese Journal of Medical Ultrasound(Electronic Edition)

基　　金：国家自然科学基金面上项目(30770562);上海市科委基础研究重点项目(10JC1412600)

摘　　要：目的探讨低强度超声照射下前列腺癌细胞的细胞膜通透性变化。方法频率为1MHz低强度超声以连续模式照射6孔板中的单层贴壁LNCaP前列腺癌细胞,以正常细胞膜非通透性荧光染料-钙黄绿素(Calcein)标记细胞膜通透性增加的细胞,以核酸荧光染料-碘化丙啶(PI)标记死亡细胞。在声强为160mW/cm2超声下照射5s,荧光显微镜观察细胞膜通透性改变,扫描电镜观察细胞膜的形态变化;分别在不同低声强(80mW/cm2、120mW/cm2和160mW/cm2)超声下照射5s,另外在声强为120mW/cm2超声下分别照射5s、10s和15s,流式细胞仪分析细胞膜通透性增加的细胞比例与声强及照射时间的对应关系。结果低强度超声作用后,荧光显微镜显示部分单层贴壁LNCaP前列腺癌细胞摄取Calcein而呈现绿色荧光,而对照组无细胞摄取Calcein;扫描电镜显示正常LNCaP前列腺癌细胞表面有密集微绒毛,而超声作用后细胞膜表面绒毛稀少、变平,少数细胞表面出现裂孔;流式细胞仪显示细胞膜通透性增加的细胞比例分别随声强和照射时间的增加而增加。结论低强度超声可以增加前列腺癌细胞膜通透性,细胞膜通透性增加的细胞表面变平,少数出现裂孔;细胞膜通透性增加的细胞比例与声强和照射时间均呈正相关。Objective To investigate the permeability changes of prostate cancer cells membrane under low intensity ultrasound in vitro. Methods The culture of monolayer adherent LNCaP prostate cancer cells in six-well plate was exposed to continuous ultrasound at frequency of 1 MHz. The cells membrane permeability （stained with Calcein）and cells viability（stained with PI）were evaluated by fluorescent microscope （FM） and cells morphological changes were evaluated by scanning electron microscope （SEM） under ultrasound with acoustic intensity of 160 mW/cm2 for 5 s. The rate of cells with increased cells membrane permeability as function of acoustic intensity （80 mW/cm2,120 mW/cm2 and 160 mW/cm2,for 5 s） and exposure duration （5 s,10 s and 15 s,acoustic intensity of 120 mW/cm2） was evaluated by flow cytometry. Results After low intensity of ultrasound,the cells with increased cell membrane permeability could be clearly shown with Calcein uptake under FM while no cell showed Calcein uptake in the control group. The SEM showed less microvilli on the cells after low intensity of ultrasound exposure and few cells showed holes on the cell membrane. The rate of cells with increased membrane permeability increased with acoustic intensity and exposure duration. Conclusion Low intensity ultrasound alone could increase membrane permeability of prostate cancer cells and cells with increased membrane permeability showed surface plane,uncommon holes on the cells membrane. The rate of cells with increased membrane permeability positively correlated with acoustic intensity and exposure duration.

关 键 词：超声检查 前列腺肿瘤 细胞膜通透性 

分 类 号：R737.25[医药卫生—肿瘤]