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通过纳米颗粒跟踪分析对源自卵巢癌细胞和正常卵巢上皮细胞的外泌体进行表征。

Characterization of exosomes derived from ovarian cancer cells and normal ovarian epithelial cells by nanoparticle tracking analysis.

作者信息

Zhang Wei, Peng Peng, Kuang Yun, Yang Jiaxin, Cao Dongyan, You Yan, Shen Keng

机构信息

Department of Obstetrics and Gynecology, Peking Union Medical College (PUMC) Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, People's Republic of China.

State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing, China.

出版信息

Tumour Biol. 2016 Mar;37(3):4213-21. doi: 10.1007/s13277-015-4105-8. Epub 2015 Oct 22.

Abstract

Cellular exosomes are involved in many disease processes and have the potential to be used for diagnosis and treatment. In this study, we compared the characteristics of exosomes derived from human ovarian epithelial cells (HOSEPiC) and three epithelial ovarian cancer cell lines (OVCAR3, IGROV1, and ES-2) to investigate the differences between exosomes originating from normal and malignant cells. Two established colloid-chemical methodologies, electron microscopy (EM) and dynamic light scattering (DLS), and a relatively new method, nanoparticle tracking analysis (NTA), were used to measure the size and size distribution of exosomes. The concentration and epithelial cellular adhesion molecule (EpCAM) expression of exosomes were measured by NTA. Quantum dots were conjugated with anti-EpCAM to label exosomes, and the labeled exosomes were detected by NTA in fluorescent mode. The normal-cell-derived exosomes were significantly larger than those derived from malignant cells, and exosomes were successfully labeled using anti-EpCAM-conjugated quantum dots. Exosomes from different cell lines may vary in size, and exosomes might be considered as potential diagnosis biomarkers. NTA can be considered a useful, efficient, and objective method for the study of different exosomes and their unique properties in ovarian cancer.

摘要

细胞外泌体参与多种疾病过程,具有用于诊断和治疗的潜力。在本研究中,我们比较了源自人卵巢上皮细胞(HOSEPiC)和三种上皮性卵巢癌细胞系(OVCAR3、IGROV1和ES-2)的外泌体的特征,以研究源自正常细胞和恶性细胞的外泌体之间的差异。使用两种既定的胶体化学方法,即电子显微镜(EM)和动态光散射(DLS),以及一种相对较新的方法,即纳米颗粒跟踪分析(NTA),来测量外泌体的大小和大小分布。通过NTA测量外泌体的浓度和上皮细胞粘附分子(EpCAM)表达。将量子点与抗EpCAM偶联以标记外泌体,并通过荧光模式下的NTA检测标记的外泌体。源自正常细胞的外泌体明显大于源自恶性细胞的外泌体,并且使用抗EpCAM偶联的量子点成功标记了外泌体。来自不同细胞系的外泌体大小可能不同,外泌体可被视为潜在的诊断生物标志物。NTA可被认为是研究卵巢癌中不同外泌体及其独特性质的一种有用、高效且客观的方法。

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