水溶性量子点在肿瘤细胞标记及荧光共振能量转移体系构建中的应用

发布时间：2018-05-01 09:20

本文选题：CdTe/CdS核/壳量子点 + 荧光探针　；参考：《东北师范大学》2012年硕士论文

【摘要】：将水溶性荧光量子点应用于肿瘤细胞成像和荧光共振能量转移这两个领域，是近年来国内外研究的热点。在本课题中尝试分别利用不同种类，不同荧光发射波长的量子点应用在这两个领域中，主要研究内容和结果如下： 1.在以单层CdTe量子点为核的基础上，通过水相合成的方法制备了最大发射波长分别为545nm和600nm的CdTe/CdS核/壳量子点。通过紫外-可见吸收光谱，荧光光谱，XRD，透射电镜等表征手段，证实了CdTe/CdS量子点的核/壳结构并且与单层CdTe量子点相比具有更强的光稳定性及荧光量子产率。通过与抗肿瘤药物5-氟尿嘧啶（5-FU），P-gp抑制剂他莫西芬(TAM)以及生物素偶联，制备了CdTe/CdS(545nm)-5-FU及bio-CdTe/CdS(600nm)-TAM两种新型荧光探针，并进一步探讨了其相互作用的机理。通过将两种荧光探针对肿瘤细胞进行标记，我们发现bio-CdTe/CdS-TAM能够靶向追踪细胞膜表面上的P-gp，并且TAM成功抑制了P-gp对抗肿瘤药物的外排作用，使抗肿瘤药物5-FU随着CdTe/CdS（545nm）进入到了细胞核内而未被排出。这使得P-gp的抑制作用能够可视化，扩大了量子点在肿瘤细胞成像和治疗领域的应用。 2.将最大发射波长分别为530nm和610nm的CdTe量子点，分别与两种生物分子——亲合素（Avi）和生物素（Bio）进行共价偶联，并分别考察了pH值，反应温度及反应物配比对偶合物的影响。借助Avi与Bio之间的强亲和力及反应的高度专一性，，成功的构建了荧光共振能量转移（FRET）体系，并考察了不同供体和受体摩尔比对FRET效率的影响，这为FRET技术在药物定量分析领域的应用奠定了基础。同时，利用高效液相色谱（HPLC）这一快速、灵敏、可靠的检测方法分析偶合物的生成及FRET的构建，大大提高了分析的准确性和灵敏性。
[Abstract]:The application of water-soluble fluorescent quantum dots in tumor cell imaging and fluorescence resonance energy transfer is a hot topic in recent years. In this paper, we try to use different kinds of quantum dots with different emission wavelengths in these two fields. The main research contents and results are as follows: 1. Based on monolayer CdTe quantum dots, CdTe/CdS core / shell quantum dots with maximum emission wavelengths of 545nm and 600nm were prepared by water phase synthesis. The core / shell structure of CdTe/CdS quantum dots was confirmed by UV-Vis absorption spectra, fluorescence spectra and transmission electron microscopy, and the photoluminescence stability and fluorescence quantum yield of CdTe/CdS quantum dots were better than those of monolayer CdTe QDs. Two novel fluorescent probes, CdTe/CdS(545nm)-5-FU and bio-CdTe/CdS(600nm)-TAM, were prepared by coupling with the anti-tumor drug 5-fluorouracil (5-FU) P-gp inhibitor tamoxifen (TAM) and biotin, and the mechanism of their interaction was further discussed. By labeling tumor cells with two fluorescent probes, we found that bio-CdTe/CdS-TAM was able to target P-gp on the surface of cell membrane, and TAM successfully inhibited the efflux of P-gp against tumor drugs. The antitumor drug 5-FU enters the nucleus with CdTe / CdSU 545 nm and is not excreted. This makes the inhibition of P-gp visible and expands the application of quantum dots in tumor cell imaging and treatment. 2. The CdTe quantum dots with the maximum emission wavelengths of 530nm and 610nm were covalently coupled with two biomolecules, avion and biotin, respectively. The effects of pH value, reaction temperature and reactant ratio on the conjugate were investigated. Based on the strong affinity between Avi and Bio and the high specificity of the reaction, the fluorescence resonance energy transfer (FRET) system was successfully constructed, and the effects of different donor and receptor molar ratios on the FRET efficiency were investigated. This laid a foundation for the application of FRET technology in the field of drug quantitative analysis. At the same time, the rapid, sensitive and reliable detection method of HPLC was used to analyze the formation of coupling compounds and the construction of FRET, which greatly improved the accuracy and sensitivity of the analysis.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.51;O657.3

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