多肽修饰的金纳米颗粒对小鼠三阴性乳腺癌细胞的靶向光热效应研究  

Peptide modified gold nanoparticles enhance the elimination of triple-negative breast cancer cells through photothermal effect

作　　者：孙博[1] 蒙艺灵 温涛 孟洁[1] 刘健[1] 许海燕[1] SUN Bo;MENG Yiling;WEN Tao;MENG Jie;LIU Jian;XU Haiyan(Institute of Basic Medical Sciences Chinese Academy of Medical Sciences,School of Basic Medicine Peking Union Medical College,Beijing 100005)

机构地区：[1]中国医学科学院基础医学研究所,北京协和医学院基础学院,北京100005

出　　处：《北京生物医学工程》2021年第5期441-446,共6页Beijing Biomedical Engineering

基　　金：中国医学科学院医学与健康科技创新工程项目(CIFMS 2016-I2M-3-004);国家自然科学基金(81801771)资助。

摘　　要：目的乳腺癌细胞高表达热休克蛋白60(heat shock protein 60,HSP60)。本文用可特异性结合的多肽P17修饰金纳米颗粒(aurum nanoparticle,AuNP),构建靶向光热治疗系统(AuNP-P17),研究其对小鼠乳腺癌细胞4T1的光热杀伤作用。方法利用紫外-可见-近红外分光光度计、动态光散射仪(dynamic light scattering,DLS)和透射电子显微镜(transmission electron microscope,TEM)对纳米颗粒的光学性质、粒径和电势进行表征;通过荧光标记和流式细胞术检测细胞HSP60表达水平;利用光学显微镜观察4T1细胞对纳米颗粒的摄取;使用便携式热电偶温度计测量细胞所在溶液在激光照射下的升温曲线;采用细胞增殖-毒性检测法(cell counting Kit-8,CCK-8)测定细胞活性。结果金纳米颗粒呈均匀球形,水合粒径为(51.8±0.1)nm;表面连接P17后(AuNP-P17)水合粒径为(52.1±2.2)nm。在金原子浓度不大于100μg/mL范围内,AuNP和AuNP-P17与4T1细胞共孵育24 h对细胞活性无显著性影响。相比于AuNP,AuNP-P17被更多地摄取到细胞中。激光照射4 min后,AuNP-P17、AuNP组和对照组的溶液温度从(22.9±0.4)℃分别上升到(52.0±3.9)℃、(47.2±0.6)℃和(38.2±2.4)℃,4T1细胞存活率分别为(1.3±2.8)%、(64.1±6.8)%和(100.0±3.2)%,两两比较差异具有统计学意义(P<0.01)。结论AuNP-P17可被4T1细胞摄取,具有良好的生物相容性、靶向性和光热效应。在激光照射下,对乳腺癌细胞具有更强的杀伤作用,在乳腺癌光热治疗方面具有转化应用潜力。Objective Breast cancer cells highly express heat shock protein 60(HSP60).In this paper,we construct a targeted photothermal therapy system by modifying aurum nanoparticle(AuNP)with a HSP60 specific binding peptide(P17)and study the photothermal killing effect on breast cancer cell(4T1).Methods The optical properties,particle size and potential of nanoparticles were characterized by UV-Vis-NIR spectrophotometer,dynamic light scattering(DLS)and transmission electron microscope(TEM).The expression of HSP60 was detected by fluorescence labeling and flow cytometry.The uptake of nanoparticles by 4T1 cells was observed by light microscope.The heating curve of cell solution under laser irradiation was measured by portable thermocouple thermometer.The cell viability was detected by cell counting kit-8(CCK-8).Results The gold nanoparticles were spherical and the hydrated particle size was(51.8±0.1)nm.The hydrated particle size of AuNP-P17 was(52.1±2.2)nm.After incubated with AuNP and AuNP-P17 for 24h,the cellular viability of 4T1 had no significant change when the concentration of gold atom was lower than 100μg/mL.The uptake of AuNP-P17 by 4T1 cells was observed much higher than that of AuNP with light microscope.With laser irradiation for 4 minutes,the temperature of 4T1 cells incubated with AuNP-P17,AuNP and the control group increased from(22.9±0.4)℃to(52.0±3.9)℃,(47.2±0.6)℃and(38.2±2.4)℃,respectively.The survival rate of 4T1 cells changed to(1.3±2.8)%,(64.1±6.8)%,and(100.0±3.2)%with significant difference(P<0.01).Conclusions AuNP-P17 can be uptake by 4T1,and has good biocompatibility,targeting and photothermal effect.With laser irradiation,it can promote the targeted killing effect of AuNP-P17 on 4T1 cells,which has the potential of transformation and application in the treatment of photothermal of breast cancer.

关 键 词：热休克蛋白60 乳腺癌细胞 光热 靶向杀伤 金纳米颗粒 

分 类 号：R318.08[医药卫生—生物医学工程]