低强度聚焦超声触发PLGA阿霉素在细胞内空化增加乳腺癌细胞死亡  

Low-intensity Focused Ultrasound-triggered Intracellular Cavitation of Doxorubicinloaded PLGA Nanoparticles Enhances Breast Cancer Cell Death

作　　者：黄盘辉 罗琼 严飞[3] 李绍林[1] HUANG Pan-hui;LUO Qiong;YAN Fei;LI Shao-lin(Department of Radiology,The Fifth Affiliated Hospital of Sun Yat-sen University,Zhuhai 519000,China;Depart-ment of Ultrasound Medicine,Guangzhou First People's Hospital,School of Medicine,South China University of Technol-ogy,Guangzhou 510180,China;Paul C.Lauterbur Research Center for Biomedical Imaging,Shenzhen Institute of Ad-vanced Technology,Chinese Academy of Sciences,Shenzhen 518055,China)

机构地区：[1]中山大学附属第五医院放射科,广东珠海519000 [2]华南理工大学医学院附属广州市第一人民医院超声科,广东广州510180 [3]中国科学院深圳先进技术研究所劳特伯生物医学成像研究中心,广东深圳518055

出　　处：《中山大学学报（医学科学版）》2021年第2期209-217,共9页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81871376)。

摘　　要：【目的】探讨低强度聚焦超声能否触发聚乳酸-羟基乙酸共聚物(PLGA)包裹阿霉素在细胞内的空化作用以增加4T1肿瘤细胞的死亡。【方法】采用双乳化溶剂蒸发法制备了负载阿霉素的聚乳酸-羟基乙酸共聚物(PLGA)纳米气泡。设立细胞内释药组,细胞外释药组,传统给药组,单独超声组和对照组。用流式细胞仪检测各组细胞的死亡情况,用细胞活力检测试剂盒(CCK8)检测各组的细胞增殖,细胞划痕检测各组的侵袭情况。【结果】超声组细胞核的红色荧光平均强度高于对照组,差异具有统计学意义(t=16.627,P<0.001);聚乳酸-羟基乙酸共聚物(PLGA)包裹阿霉素在3、5、7、24和48 h的累计释药量超声组均高于对照组,差异都具有统计学意义(P<0.001);细胞内释药组,细胞外释药组及传统给药组的细胞存活率,细胞增值率及划痕像素面积均低于对照组,且细胞内释药组显著低于其他组,差异具有统计学有意义(P<0.001)。【结论】低强度聚焦超声通过细胞内空化触发的细胞内药物释放比低强度聚焦超声触发的细胞外药物释放及传统的给药方式更有效地增加4T1肿瘤细胞的死亡。【Objective】To investigate whether low-intensity focused ultrasound can trigger the intracellular cavitation of the doxorubicin-loaded polylactic acid-glycolic acid copolymer(PLGA)nanoparticles and then enhance the death of 4T1 breast cancer cells.【Methods】The doxorubicin-loaded PLGA nano-bubbles were prepared by double emulsification solvent evaporation method.4T1 breast cancer cells were divided into intracellular drug release group,extracellular drug release group,traditional drug delivery group,ultrasound alone group and control group.In each group,flow cytometry,cell counting kit-8(CCK8)and scratch assay were used to detect the cell death,proliferation and invasion,respectively.【Results】The average intensity of red fluorescence in the nuclei of the ultrasound group was higher than that of the control group and the difference was statistically significant(t=16.627,P<0.001).The cumulative drug release of PLGA encapsulated doxorubicin at 3,5,7,24 and 48 h in the ultrasound group were all higher than those in the control group and the differences were statistically significant(P<0.001).The cell viability,cell proliferation and 24 h scratch pixel area in the intracellular drug release group,extracellular drug release group and traditional drug delivery group were all lower than those in the control group,with the results in the intracellular drug release group significantly lower than those in other groups.All the differences were statistically significant(P<0.001).【Conclusions】The intracellular drug release triggered by low-intensity focused ultrasound through intracellular cavitation is more effective in enhancing the death of 4T1 tumor cells than the extracellular drug release triggered by low-intensity focused ultrasound and traditional drug delivery methods.

关 键 词：低强度聚焦超声 细胞内空化 乳腺肿瘤 细胞死亡 

分 类 号：R73-361[医药卫生—肿瘤]