北京生物医学工程

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 enhancethe elimination of triple-negative breast cancercellsthrough photothermal effect

作者：孙博蒙艺灵温涛孟洁刘健许海燕

单位：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） 通信作者：许海燕，E-mail：xuhy@pumc.edu.cn；温涛，E-mail：went@ibms.pumc.edu.cn

关键词：热休克蛋白60；乳腺癌细胞；光热；靶向杀伤

分类号：R318.04

出版年·卷·期（页码）：2021·40·5（441-446）

摘要：

目的乳腺癌细胞高表达热休克蛋白60（HSP60）。本文用可特异性结合的多肽P17修饰金纳米颗粒（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100 μg/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）°C分别上升到（52.0 ± 3.9）°C、（47.2 ± 0.6）°C和（38.2 ± 2.4）°C，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 gold 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 are spherical and the hydrated particle size is (51.8 ± 0.1) nm. The hydrated particle size of AuNP-P17 is (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 100 μg/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) °C to (52.0 ± 3.9) °C, (47.2 ± 0.6) °C and (38.2 ± 2.4) °C, 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.

参考文献：

1.Wild?CP, Weiderpass?E, Stewar?BW. World cancer report: cancer research for cancer prevention [R]. Lyon，Geneva：World Health Organization，2020.

2. Harbeck N, Gnant ?M. Breast cancer [J]. The Lancet，?2017，389(10074):1134-1150.

3. Lee YT, Tan YJ, Oon CE. Molecular targeted therapy: Treating cancer with specificity?[J].?European Journal of Pharmacology，?2018，834:188-196.

4. Colli LM, Machiela MJ, Zhang H,et al. Landscape of combination immunotherapy and targeted therapy to improve cancer management [J].?Cancer?Research，2017，77(13):3666-3671.

5. ????Valachis A,?Polyzos NP,?Alpha NP,?et al. Bevacizumab in metastatic breast cancer:a meta-analysis of randomized controlled trials?[J].?Breast Cancer Research and Treatment，?2010，122(1):1-7．

6. ????Litton JK,?Rugo HS,?Ettl J,?et al. Talazoparib in patients with advanced breast cancer and a germline BRCA mutation?[J].?New England Journal of Medicine，2018，379(8):753-763．

7. ????Robson M,?Im SA,?Senkus E,?et al.?Olaparib for metastatic breast cancer in patients with a germline BRCA mutation?[J].?New England Journal of Medicine，?2017，377(6):523-533．

8. Chen FM, Huang LJ, Ou-Yang F,et al. Activation of mitochondrial unfolded protein response is associated with Her2-overexpression breast cancer [J]. Breast Cancer Research and Treatment，?2020，183(1):61-70.

9. Bini L, Magi B, Marzocchi B, et al. Protein expression profiles in human breast ductal carcinoma and histologically normal tissue [J]. Electrophoresis，1997，18(15):2832-2841.

10. Buttros DdAB, Branco MT, Orsatti CL,et al. High risk for cardiovascular disease in postmenopausal breast cancer survivors [J]. Menopause，?2019，26(9):1024-1030.

11. Yang S, Meng J, Yang Y, et al. A HSP60-targeting peptide for cell apoptosis imaging [J]. Oncogenesis，?2016，5:e201.

12. Bastus NG, Comenge J, Puntes V. Kinetically controlled seeded growth synthesis of citrate-stabilized gold nanoparticles of up to 200 nm: size focusing versus Ostwald ripening [J]. Langmuir，2011，27(17):11098-11105.

13. Merendino AM, Bucchieri F, Campanella C, et al. Hsp60 is actively secreted by human tumor cells [J]. PLoS One，?2010，5(2):e9247.

14. Kim M, Lee JH, Nam JM. Plasmonic photothermal nanoparticles for biomedical applications [J]. Advanced?Science (Weinh)，?2019，6(17):1900471.

15. Gupta N, Malviya R. Understanding and advancement in gold nanoparticle targeted photothermal therapy of cancer [J]. Biochimica et?Biophysica?Acta-Reviews on?Cancer，2021，1875(2):188532.

服务与反馈：

【文章下载】【加入收藏】

提示：您还未登录，请登录！点此登录