乳腺癌细胞及其外泌体的拉曼表型快速鉴定与关系研究

doi:10.3964/j.issn.1000-0593(2023)12-3840-07

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Abstract Exosomes are nano-sized phospholipid bilayer-enclosed vesicles secreted by all cells into the extracellular milieu. Released exosomes contain cell-specific proteins, membrane lipids, mRNA, DNA and microRNA that can perform versatile roles in normal or diseased processes. Exosomes are ideal biomarkers of cancer, which have important application potential in liquid biopsy and are expected to become one of the means for rapid cancer detection. Surface enhanced Raman spectroscopy (SERS) is a molecular vibration spectrum, which can detect the fine structure and information changes of substances at the molecular level, and has the characteristics of a “fingerprint spectrum”. In this study, the exosomes were isolated via differential centrifugation combined with ultracentrifugation of the supernatants of breast cells. The SERS profiles of breast cancer cells and their exosomes were collected with colloidal Au nanoparticles as the enhanced substrate, and multivariate statistical analysis was used to identify and distinguish breast cancer cells. The results showed that breast cancer cells and their exosomes had characteristic Raman signals in the range of 500～1 600 cm^-1. Their Raman phenotypes obtained by non-labeled detection were the presentation of all the signals of the“whole-organism fingerprint” of the sample. The accuracy rate reached 100% by using exosome-SERS detection and OPLS-DA analysis. Single-cellular SERS detection combined with PCA-LDA analysis showed that the accuracy of differentiating breast cancer cells was 83.7%. Breast cancer cells and their exosomes showed similarity at the bands of 506～569 and 1 010～1 070 cm^-1, but the characteristic Raman peaks of exosomes at 735, 963 and 1 318 cm^-1 were significantly higher than those of cells. It may be because the structure of exosomes is simpler than that of cells, and the information of biological macromolecules such as nucleic acids and proteins can be characterized more easily, indicating the feasibility of rapid identification of breast cancer by detecting exosomes by SERS technology. In summary, this study established a non-labeling and direct detection method for rapidly detecting single cells and their exosomes by SERS analysis. Combined with multivariate statistical analysis, different types of breast cancer cells could be quickly identified, and the relationship between exosomes and maternal cells was explored from the perspective of Raman omics. This method has the advantages of non-labeling, rapidness, sensitivity, accuracy and simplicity, which provides an effective technical means for rapid diagnosis and screening of breast cancer in vitro, and lays a foundation for clinical application.

Key words：Surface enhanced Raman spectroscopy;Breast cancer;Exosome;Single-cellular analysis;Rapid detection;Multivariate statistical analysis

Received: 2022-07-11 Accepted: 2022-10-17

ZTFLH:

Q28，R319

Corresponding Authors: ZHANG Ping E-mail: zplife@bjut.edu.cn

Cite this article:

LU Wen-jing,FANG Ya-ping,LIN Tai-feng, et al. Rapid Identification of the Raman Phenotypes of Breast Cancer Cell Derived Exosomes and the Relationship With Maternal Cells[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(12): 3840-3846.

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https://www.gpxygpfx.com/EN/10.3964/j.issn.1000-0593(2023)12-3840-07 OR https://www.gpxygpfx.com/EN/Y2023/V43/I12/3840

[1] Guillaume van Niel, Gisela D'Angelo, Graça Raposo. Nature Reviews Molecular Cell Biology, 2018, 19(4): 213.
[2] Johan Skog, Tom Würdinger, Sjoerd van Rijn, et al. Nature Cell Biology, 2008, 10(12): 1470.
[3] Wu Di, Chen Yonghao, Hou Shuai, et al. ChemBioChem: A European Journal of Chemical Biology, 2019, 20(19): 2432.
[4] Chenchen Fan, Na Zhao, Kai Cui, et al. ACS Sensors, 2021, 6(9): 3234.
[5] Li Juan, Li Yanru, Chen Shuai, et al. Small Methods, 2022, 6(6): e2200154.
[6] Jelle Penders, Anika Nagelkerke, Eoghan M Cunnane, et al. ACS Nano, 2021, 15(11): 18192.
[7] Li Guohao, Zhu Nanhang, Zhou Juan, et al. Journal of Materials Chemistry B, 2021, 9(11): 2709.
[8] Wang Yanlin, Li Qiaoyu, Shi Haimei, et al. Lab on A Chip, 2020, 20(24): 4632.
[9] Wei Xiaodan, Zheng Dawei, Zhang Ping, et al. Journal of Applied Biomaterials and Fundamental Materials, 2018, 16: 157.
[10] Zhang Ping, Wang Limin, Fang Yaping, et al. Molecules, 2019, 24(16): 2947.
[11] Gergo Peter Szekeres, Janina Kneipp. Frontiers in Chemistry, 2019, 31(7): 30.
[12] Wei E Huang, Robert I Griffiths, Ian P Thompson, et al. Analytical Chemistry, 2004, 76(15): 4452.
[13] David Perez-Guaita, Kamila Kochan, Miguela Martin, et al. Vibrational Spectroscopy, 2017, 91: 46.
[14] Christian Matthäus, Tatyana Chernenko, Judith A Newmark, et al. Biophysical Journal, 2007, 93(2): 668.
[15] Andrade P O, Bitar R A, Yassoyama K, et al. Analytical and Bioanalytical Chemistry, 2007, 387(5): 1643.
[16] Nicholas Stone, Catherine Kendall, Jenny Smith, et al. Faraday Discussions, 2004, 126: 141.
[17] Han H W, Yan X L, Dong R X, et al. Applied Physics B: Lasers and Optics, 2009, 94(4): 667.
[18] Nicholas Stone, Pela Stavroulaki, Catherine Kendall, et al. Laryngoscope, 2010, 110(10): 1756.
[19] Li Xiaozhou, Yang Tianyue, Li Siqi, et al. Optics Express, 2015, 23(14): 18361.
[20] Tang Hong-wu, Xuebin B Yang, Jennifer Kirkham, et al. Analytical Chemistry, 2007, 79(10): 3646.